CN104961782B - Bioactive ingredients and its medical usage in Green Bamboo Leaf Liquor - Google Patents

Bioactive ingredients and its medical usage in Green Bamboo Leaf Liquor Download PDF

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CN104961782B
CN104961782B CN201510312849.7A CN201510312849A CN104961782B CN 104961782 B CN104961782 B CN 104961782B CN 201510312849 A CN201510312849 A CN 201510312849A CN 104961782 B CN104961782 B CN 104961782B
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bamboo leaf
green bamboo
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liver
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CN104961782A (en
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王金辉
李国玉
黄健
高红英
杜小威
韩英
王书云
安丽娟
王立飞
程锁明
孙富周
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Shihezi University
Shanxi Xinghuacun Fen Wine Factory Co Ltd
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Shanxi Xinghuacun Fen Wine Factory Co Ltd
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Abstract

The present invention relates to the bioactive ingredients in Green Bamboo Leaf Liquor and health-care effect, in particular to the bioactive ingredients in Green Bamboo Leaf Liquor with immunoregulatory activity, anti-oxidant, anti-inflammatory, protection hepar damnification.

Description

Bioactive ingredients and its medical usage in Green Bamboo Leaf Liquor
Technical field
The present invention relates to the bioactive ingredients in Green Bamboo Leaf Liquor and health-care effect, in particular to having in Green Bamboo Leaf Liquor Immunoregulatory activity, anti-oxidant, anti-inflammatory, the bioactive ingredients for protecting hepar damnification.
Background technology
Green Bamboo Leaf Liquor is China's traditional history famous brand of wine, and Chinese win high praise highest and most health liquors.It is with China Delicate fragrance type famous brand of wine --- Fenyang wine is base liquor, with the pleasant impression rare Chinese medicine of the leaf of bamboo, Radix Angelicae Sinensis, dried orange peel, cape jasmine, fructus amomi, santal, cloves etc. ten A kind of alcoholic drink mixed with fruit juice that the soak and rock sugar of material are formulated.The drinking utensils has blood-nourishing and stomach, helped digestion, relieving restlessness and other effects.It is throughout the year suitable Amount is drunk, can reconcile internal organs, dredge gas blood-nourishing, the fiery dissolving phlegm that disappears, removing toxic substances diuresis, invigorating the spleen nourishing liver.Not only can may be used also with health of body To prevent diseases such as curing arthritis, hypertension, highs fat of blood.
In recent years, with the continuous enhancing of consumer health's health care consciousness, special efficacy composition for health food and Its health-care effect, also increasingly cause the concern of people.Therefore, it is deep to bioactive ingredients progress system in Green Bamboo Leaf Liquor Research, attempt to excavate, inquire into its distinctive health function, there is important theory significance and actual application value.
The content of the invention
The purpose of the present invention：Chemical composition in the research Green Bamboo Leaf Liquor of systematic science, finds new activated monomer chemical combination Thing.
The implementation process of invention is as follows：
Include silica gel column chromatography, macroporous absorbent resin, polyamide, sephadex lh-20 post with various separation means Chromatogram and preparative high performance liquid chromatography etc., isolated 96 compounds from Green Bamboo Leaf Liquor, wherein 16 terpenoids Thing, 10 iridoids, 25 flavones and flavonoid glycoside compound, 3 chromogen ketone compounds, 15 phenolic acid and Phenol glycosides compound, 14 aromatic compounds, 1 chinic acid class, 2 Coumarinses, 2 lignanoids, 3 steroids, 2 Individual long-chain fat acids, 2 furfural classes, 1 Furanones.Wherein compound 1,2,3,4,5,14,16,53 for new chemistry into Point.
Its chemical name of these compounds and structural formula are as follows：
Compound 1：The ring [5.4.0] 11 of (1R, 10S, 11R) -10,11- dimethyl -4- aldehyde radical -2,9- dioxos-two - 4,6- diene -3- ketone, [(1R, 10S, 11R) -10,11-dimethyl-4-formyl-2,9-dioxa-bicyclo [5.4.0] Undeca-4,6-dien-3-one], there is structure shown in structural formula 1.
Compound 2：6- (6- (3,4- dimethoxys) phenylpropenoyl-β-D-Glucose base)-O- β-D-Glucose first glycosides [methyl-6-(6-(3,4-dimethoxy)-benzalacryloyl-β-D-glucosyl)-O-β-D- Glucopyranoside], there is structure shown in structural formula 2.
Compound 3：(4- hydroxyl -2,6,6- trimethyls)-cyclohexenecarboxylic acid -4- hydroxyls-(benzoic acid) ester [Picrocrocinic ester], there is structure shown in structural formula 3.
Compound 4：(7R) -6 '-O- (3 "-methoxyl group -4 "-hydroxyl-phenylpropenoyl)-β-- O-7- the hydroxyls of D-Glucose -1 ' Methyl-(6- methylol -1,1- dimethyleyelohexane -4- alkene -3- ketone) glycosides [6'-O- (3-methoxyl-4-hydroxyl- Coumaroyl)-epijasminoside B], there is structure shown in structural formula 4.
Compound 5：(4R) -3- methylol -4- methylol -5,5- dimethyleyelohexane -2- ketenes-β-D-Glucose glycosides [epijasminoside B], there is structure shown in structural formula 5.
Compound 14：(5R)-(2E) -5- hydroxy-2-methyls-amyl- 2- alkene -1,6- diketone [(5R)-(2E) -5- Hydroxy-2-methyl-hepta-2-ene-1,6-dione], there is structure shown in structural formula 6.
Compound 16：Trans-p- methoxyl groups of 6 "-O--cinnamyl group Geniposide O-gentibioside [6 "-O-trans-p- Methoxyl-coumaroylgenipin gentiobioside], there is structure shown in structural formula 7.
Compound 53：7- methoxyl groups-Isobiflorin [7-methoxyl-isobiflorin], there is the institute of structural formula 8 Show structure.
It is isolated from Green Bamboo Leaf Liquor it is another object of the present invention to provide Green Bamboo Leaf Liquor involved in the present invention Compound medical usage.
The present invention has found that Green Bamboo Leaf Liquor, isolated compound has anti-oxidant from Green Bamboo Leaf Liquor by research, resists Inflammation, Immune-enhancing effect, protection liver effect.It can be used for preparing medicine or health food of the prevention and treatment with above-mentioned effect.
The present invention also provides the medicine or health care food of isolated compound containing Green Bamboo Leaf Liquor or from Green Bamboo Leaf Liquor Product composition, to be adapted to application.
Its suitable form of described pharmaceutical composition includes any dosage form suitably taken, such as the present invention Pharmaceutical composition, can be prepared into pharmaceutical dosage forms, such as oral dosage form as needed when in use, injection form, External preparation form, suppository form etc..
The health-care food composition, including but not limited to following food form：Beverage, dairy products, bread, cake, sugar Fruit etc..
Preferably, the present invention provides above-mentioned 8 kinds of compounds, their preparation, and they are preparing medicine or health care food Application in product.
The present invention has found that Green Bamboo Leaf Liquor has anti-oxidant, anti-inflammatory, Immune-enhancing effect, protects liver effect by research, wherein Terpene, iridoids, flavonoids and phenolic acid compound are its principle active component.Bioactivity research shows, compound Any compound has anti-oxidant, anti-inflammatory, removes free radical, anticholinesterase, Immune-enhancing effect, protection liver effect in 1-98, It is the material base that Green Bamboo Leaf Liquor plays healthcare function.
The implementation process of invention is as follows：
Establish acute hepatic injury model (chemical liver injury, Immune liver injury, the wine of different damage mechanisms Essence liver injury model, liver lesion induced by drugs wound model), the research of Hepatocyte protection has been carried out to Green Bamboo Leaf Liquor.Establish immunity Low model, study the immunoregulatory activity of Green Bamboo Leaf Liquor.The cellular damages of HepaG 2 are induced using alcohol, screening liver cell protects Protect composition.The anti-inflammatory activity of compound in Green Bamboo Leaf Liquor is studied using LPS inductions RAW 264.7.To getting in Green Bamboo Leaf Liquor Compound carries out the screening of cholinesterase activity.
Brief description of the drawings
CD the and UV collection of illustrative plates of Fig. 1 compounds 1
CD the and UV collection of illustrative plates of Fig. 2 compounds 2
CD the and UV collection of illustrative plates of Fig. 3 compounds 3
CD the and UV collection of illustrative plates of Fig. 4 compounds 4
CD the and UV collection of illustrative plates of Fig. 5 compounds 7
Fig. 6 CCl4Hepatic tissue pathology change HE dyeing (100 ×) (A) normal groups of caused acute liver； (B)CCl4Model group；(C)-(F) is respectively green bamboo snake mother liquor A-D dosage groups
Hepatic tissue pathology change HE dyeing (100 ×) (A) normal groups of acute liver caused by Fig. 7 TAA；(B) TAA model groups；(C)-(F) is respectively green bamboo snake mother liquor A-D dosage groups；(G) Compound Liver-benepitino Remedy group (200mg/kg)
Hepatic tissue pathology change HE dyeing (× 100) (A) normal groups of acute liver caused by Fig. 8 alcohol； (B) alcohol model group；(C)-(F) is respectively Green Bamboo Leaf Liquor mother liquor A-D dosage groups；(G) it is DDB group (150mg/kg)
Acute liver hepatic tissue TNF-α expression (× 200) (A) normal group caused by Fig. 9 alcohol；(B) alcohol mould Type group；(C)-(F) is respectively Green Bamboo Leaf Liquor mother liquor A-D dosage groups；(G) it is DDB group (150mg/kg)
Acute liver hepatic tissue Fas expression (× 200) (A) normal groups caused by Figure 10 alcohol；(B) alcohol mould Type group；(C)-(F) is respectively Green Bamboo Leaf Liquor mother liquor A-D dosage groups；(G) it is DDB group (150mg/kg)
Acute liver hepatic tissue FasL expression (× 200) (A) normal groups caused by Figure 11 alcohol；(B) alcohol mould Type group；(C)-(F) is respectively Green Bamboo Leaf Liquor mother liquor A-D dosage groups；(G) it is DDB group (150mg/kg)
Acute liver hepatic tissue Bcl-2 expression (× 200) (A) normal groups caused by Figure 12 alcohol；(B) alcohol Model group；(C)-(F) is respectively Green Bamboo Leaf Liquor mother liquor A-D dosage groups；(G) it is DDB group (150mg/kg)
Acute liver hepatic tissue Bax expression (× 200) (A) normal groups caused by Figure 13 alcohol；(B) alcohol mould Type group；(C)-(F) is respectively Green Bamboo Leaf Liquor mother liquor A-D dosage groups；(G) it is DDB group (150mg/kg)
The extraction flow chart of Figure 14 Green Bamboo Leaf Liquors
Figure 15 Green Bamboo Leaf Liquor HPD10060% ethanol elutions are partially separated flow chart
Figure 16 Green Bamboo Leaf Liquors chloroform and ethyl acetate extraction part separation process figure
The related figures of the HMBC of Figure 17 compounds 1
The relative configuration figure of Figure 18 A compounds 1, the relative configuration figure of Figure 18 B compounds 1
The HMBC figures of Figure 19 A compounds 2, the NOEs figures of Figure 19 B compounds 2
The related figures of the HMBC of Figure 20 A compounds 3, the related figures of HMBC of Figure 20 B compounds 3, the HMBC of Figure 20 C compounds 3 Correlation figure
The related figures of the HMBC of Figure 21 A compounds 4, the related figures of HMBC of Figure 21 B compounds 4
The related figures of the HMBC of Figure 22 A compounds 5, the related figures of HMBC of Figure 22 B compounds 5
The related figures of the HMBC of Figure 23 compounds 14
The related figures of the HMBC of Figure 24 compounds 16
Embodiment
The present invention is further illustrated by the following examples, but not as limitation of the present invention.
Embodiment 1：The separation of compound
After taking Green Bamboo Leaf Liquor mother liquor, concentration to fling to alcohol, extracted successively with petroleum ether, chloroform, ethyl acetate, extract 5 respectively Time, each extract layer solvent is reclaimed, obtains each extract layer medicinal extract：Petroleum ether layer 100g, chloroform layer 56g, ethyl acetate layer 100g.
Water layer HPD after extraction100Macroporous absorbent resin is handled, and is entered successively with water, 60% ethanol, 95% ethanol Row elution, wherein water-wash section discards, and 60% ethanol is separately recovered, 95% ethanol elution part obtains medicinal extract：60% ethanol elution Part 200g, 95% ethanol elution part 20g.Extraction separation process is shown in Figure 14.
Chloroform layer, ethyl acetate layer sample are merged, sample 100g after merging is taken, utilizes silica gel column chromatography repeatedly, polyamides 50 compounds are obtained in the separation of the means such as amine, Sephadex LH-20 and HPLC；Take HPD100Macroporous absorbent resin 60% EtOH elution fraction sample 100g, utilize isolated 48 compounds of means such as silica gel column chromatography, open ODS and HPLC.Point See Figure 15 from flow, 16.
Wherein 8 noval chemical compounds of the invention, compound 1,2,3,4,5,14,16,53 specific separation method descriptions are such as Under：
Ethyl acetate fraction (100g) is with petroleum ether/acetone (100:0-0:100) silicagel column is carried out for eluting solvent Chromatograph (350g, 200-300 mesh, 9 × 160cm), fraction A-O must be eluted.Fraction I (petroleum ether/acetone, 100:15,2.3g) and Fraction M (petroleum ether/acetone, 100:50,3.0g) merging carries out half preparation HPLC separation (acetonitrile/water, 15/85) and obtains compound 3 (18mg)。
60% alcohol elution (100g) is with chloroform/methanol (100:0-0:100) silica gel column layer is carried out for eluting solvent Analyse (350g, 200-300 mesh, 9 × 160cm), fraction a-m must be eluted.Fraction b (chloroform/methanol, 100:0.5,70mg) with oil Ether/acetone carries out silica gel column chromatography, obtains fraction b1-b3.Fraction b2, which carries out half preparation HPLC separation (methanol/water, 4/96), to be changed Compound 6 (3.0mg).
Fraction c (chloroform/methanol, 100:2,2.0g) carry out half preparation HPLC separation (acetonitrile/water, 6/94) and obtain compound 1 (5.1mg).Fraction e (chloroform/methanol, 100:3 and 100:5) with methanol/water (15:85-40:60, v/v) ODS gradients are carried out to wash Take off to obtain five parts, e1-e5.Fraction e1 (methanol/water, 15:85,38.9mg) carry out half prepare HPLC separation (acetonitrile/water, 20/ 80) compound 2 (3.5mg) is obtained.
Fraction f (chloroform/methanol, 100:8,3.0g) with methanol/water (5:95-50:50, v/v) ODS gradient elutions are carried out to obtain Six part f1-f6.Fraction f2 (methanol/water, 10:90,52.6mg) half preparation HPLC separation (acetonitrile/water, 5/95) is carried out to obtain Compound 4 (4.2mg), compound 5 (3.0mg) and compound 8 (8.5mg).
Fraction g (chloroform/methanol, 100:12) silica gel column chromatography is carried out as eluting solvent using chloroform/methanol and obtains three fractions g1-g3.Fraction g2 carries out half preparation HPLC separation (acetonitrile/water, 21/79) and obtains compound 7 (4.5mg).
Embodiment 2：Identification of chemical structure
Using 1 dimension, 2 dimension nuclear magnetic resoance spectrums (1D, 2D-NMR), mass spectrum (MS), the spectrum means such as circular dichroism spectra (CD) and Other physico-chemical processes determine the chemical constitution of 96 isolated compounds.Including 8 noval chemical compounds Chemical constitution and identification of means such as table 1：
The structure and its authentication method of 96 compounds (including 8 noval chemical compounds) in the Green Bamboo Leaf Liquor of table 1
* it is noval chemical compound
Embodiment 3：The Identification of chemical structure of compound 1
Yellow transparent solid body (methanol), is dissolved in methanol.HR-ESI-TOF-MS spectrums provide high-resolution quasi-molecular ion peak m/ z223.0973[M+H]+(Calcd.223.0970,1.3ppm).With reference to its NMR data, its molecular formula C is determined12H14O4, and count Calculate the compound and contain 6 degrees of unsaturation.
1H H NMR spectroscopies (600MHz, CD3OD), low field area provides 2 alkene Hydrogen Proton signal δ 6.35 (1H, d, J=4.2Hz), 7.15 (1H, d, J=4.2Hz)；1 active hydrogen proton signal δ 9.33 (1H, s).High field region provides 2 company's oxygen tertiary carbon proton letters Number δ 5.23 (1H, d, J=11.4Hz), 4.34 (1H, dq, J=3.0,6.0Hz)；1 company oxygen secondary carbon proton signal δ 4.65 (2H, s)；1 tertiary carbon proton signal δ 2.65 (1H, m), two methyl proton signal δ 1.13 (3H, d, J=6.0Hz), 1.55 (3H, d, J =6.0Hz).
13C H NMR spectroscopies (150MHz, CD3OD 12 carbon signals are provided in), low field area provides 6 sp2The carbon signal of hydridization, An aldehyde radical carbon signal is wherein provided at δ 180.5, is the carbonyl carbon signals of one one-tenth ester at δ 174.6.δ 111.9,128.3,133.4, Two double bond carbon signals are provided at 146.3.Two company's oxygen methine carbon signals are provided at high field region δ 82.3,63.8, at δ 56.8 1 company's Oxymethylene carbon signal is provided, 1 methine carbon signal is provided at δ 45.3, two methyl carbon are provided at δ 14.7,18.8 Signal.
In HMBC spectrums (Figure 17), by δ 1.13 (3H, d, J=6.0Hz), 1.55 (3H, d, J=6.0Hz) respectively with δ 45.3,63.8,82.3 have correlation, and δ 4.34 (1H, dt, J=3.0,6.6Hz) is related to δ 14.7, δ 2.65 (1H, m) and δ 14.7,18.8,63.8,82.3 is related, illustrates that 14.7 methyl is connected on 82.3 company's oxygen tertiary carbon, 18.8 methyl is connected in 45.3 Tertiary carbon on.There are long-range correlation, δ 5.23 (1H, d, J=11.4Hz) and δ 14.7 by δ 4.65 (2H, s) and δ 111.9,146.3, 45.3,133.4,146.3,174.6 is related, and δ 6.35 (1H, d, J=4.2Hz) and δ 56.8,128.3,133.4,146.3 is related, δ 7.15 (1H, d, J=4.2Hz) and δ 111.9,133.4,146.3,180.5 is related, can draw the planar structure of the compound such as Fig. 1.In addition, being J=11.4Hz according to H-10/H-11 coupling constant, H10/H-9 coupling constant is J=3.0Hz, and profit With computer model configuration least energy (CS Chem 3D Pro Version 8.0, MM2minimize energy Caculate) calculate, relative configuration such as Figure 18 of the compound can be drawn, be named as 10,11- dimethyl -4- aldehyde radicals -2,9- bis- - 4,6- diene -3- the ketone of the ring of oxo-two [5.4.0] 11.
In CD is composed (see accompanying drawing 1), according to the spiral of unsaturated lactone rule, compound 1 is presented just at 306nm Cotton effects, negative cotton effects are presented at 260nm, can determine whether to meet P- spirals, therefore judge 1 for R types, determine compound 1 absolute configuration is (1R, 10S, 11R).Retrieved through system documentation, to have no the noval chemical compound of document report, its structure is - 4,6- diene -3- the ketone of-two ring [5.4.0] of (1R, 10S, 11R) -10,11- dimethyl -4- aldehyde radical -2,9- dioxos 11.
The compound 1 of table 21H NMR(600MHz in CD3OD),13C NMR(150MHz in CD3) and HMBC data OD
Embodiment 4：The Identification of chemical structure of compound 2
White powder (methanol), is soluble in methanol, insoluble in petroleum ether, ethyl acetate, chloroform.HR-ESI-TOF-MS is composed Provide high-resolution quasi-molecular ion peak m/z 545.1877 [M-H]-(Calcd.545.1870,0.5ppm).With reference to its NMR number According to it is C to determine its molecular formula24H34O14, and calculate the compound and contain 8 degrees of unsaturation.
1H H NMR spectroscopies (400MHz, CD3OD 1 group 1 " ' is provided in), 3 " ', 4 " '-trisubstituted benzene ring proton signal δ 6.90 (2H, S, H-2 " ', 6 " '), δ 7.48 (1H, s, H-5 " '), 2 with carboxyl conjugation trans double bond proton signal δ 6.38 (1H, d, J= 16.0Hz, H- α), δ 7.59 (1H, d, J=16.0Hz, H- β), 3 methoxy proton signal δ 3.71 (3H, s), δ 3.88 (3H, S), δ 3.86 (3H, s), by β-D-Glucose anomeric proton signal δ 4.98 (1H, d, J=8.0Hz, glc-1) and acylated Portugal Grape 6 proton signal δ 4.26 (1H, br.d, J=14.4Hz, glc-6) of sugar, δ 4.18 (1H, br.d, J=14.4Hz, glc-6) And by another β-D-Glucose anomeric proton signal δ 4.71 (1H, d, J=8.0Hz, glc-1) and the grape to low field displacement Sugared 6 proton signal δ 4.47 (1H, dd, J=11.2,6.0Hz, glc-6), δ 4.39 (1H, dd, J=11.2,2.4Hz, glc- 6), thus it is speculated that the glucose 6 is substituted by glucosyl group, and forms first glycosides.
13C H NMR spectroscopies (100MHz, CD3OD provided in) and share 24 carbon signals, low field shows 9 sp2The carbon letter of hydridization Number, wherein δC168.9 be carbonyl carbon signals.With reference to1H H NMR spectroscopies speculate that wherein 11 carbon signals are shown as 3 " ', 4 " '-dimethoxy Base benzene acryloyl structural framework, remaining 13 carbon signal are shown as carbon signal on two β-D-Glucoses and methoxyl group Carbon signal.
δ is understood by HSQC spectrumC51.9 (C-1), 57.0 (C-3 " '), 57.0 (C-4 " ') are three methoxyl group carbon signals, δC61.9 (C-6 "), 64.2 (C-6') two secondary carbon signals are respectively six carbon signals on two β-D-Glucoses, δC 100.6 (C-1'), 99.4 (C-1 ") two tertiary carbon signals are respectively the end group carbon signal on two β-D-Glucoses, δC 145.0 (C- β), 115.8 (C- α) are respectively and the carbon signal in the trans double bond of carboxyl conjugation, δC128.5 (C-1 " '), 149.5 (C- 3 " '), 149.4 (C-4 " ') are the nuclear substituted quaternary carbon signal of benzene, δC106.9 (C-2 " ', 6 " '), 112.4 (C-5 " ') are 3 " ', three unsubstituted carbon signals in 4 " '-dimethoxy benzene acryloyl structural framework.
In HMBC spectrum (Figure 19), methoxyl group proton δH3.65 with δC100.6 (C-1') are related, infer that C-1 is connected to On the end group of β-D-Glucose, first glycosides is formed.Methoxyl group proton δH3.86 and 3.88 respectively with δC149.4 (C-4 " '), 149.5 There is long-range correlation in (C-3 " '), illustrate that the two methoxyl groups are connected on phenyl ring C-4 " ', C-3 " ' positions.6 matter of glucose Subsignal δH4.39 with δC99.4 (C-1 ") have correlation, show that 6 of this glucose are connected with 1 of another β-D-Glucose. 6 proton signal δ of another β-D-GlucoseH4.26 with δC168.9 (C-9 " '), 115.8 (C-8 " ') are related, that is, illustrate this Glucose 6 is connected on phenylpropenoyl.In addition, phenyl ring proton signal δH6.90 with δC128.5 (C-1 " '), 149.5 (C- 3 " ') long-range correlation be present in, 149.4 (C-4 " '), 145.0 (C-7 " ').
In NOESY spectrum (Figure 19), H-1/H-1' is related, it is known that the end group of β-D-Glucose is substituted by methyl, is formed First glycosides.H-6'/H-1 " is related, illustrates that 6 of this β-D-Glucose are connected with the end group of another glucose.H-6 "/H-8 " ', H- 6 "/H-2 " ' is related, shows that this glucose 6 is connected with phenylpropenoyl.In addition, H-8 " '/H-2 " ', H-7 " '/H-6 " ' phase Close, NOESY effects can combine simultaneously1H NMR coupling constant judges C- α, and C- β double bonds are trans double bond.
Therefore, to sum up analyze, can determine that the compound is 6- (6- (3,4- dimethoxy) phenylpropenoyl-β-D- grapes Glycosyl)-O- β-D-Glucose first glycosides.
The compound 2 of table 31H NMR(400MHz in CD3OD),13C NMR(100MHz in CD3OD),HMBC,NOESY Spectrum
Embodiment 5：The Identification of chemical structure of compound 3
Yellow powder (methanol), is dissolved in methanol.HR-ESI-TOF-MS spectrums provide high-resolution quasi-molecular ion peak m/z 303.1225[M-H]-(Calcd.303.1232,-1.9ppm).With reference to its NMR data, it is C to determine its molecular formula17H20O5, and Calculate the compound and contain 8 degrees of unsaturation.
1H H NMR spectroscopies (600MHz, CD3OD in), low field area provides the phenyl ring proton signal δ of one group of AA'BB' Coupling System 6.84 (2H, dd, J=2.4,8.4Hz), 6.84 (2H, dd, J=2.4,8.4Hz).High field region provides 3 methyl proton signal δ 1.09 (3H, s), 1.21 (3H, s), 1.72 (3H, s).Two methene proton signal δ 1.74 (1H, m), 1.40 (1H, m), 1.95 (1H, m), 2.33 (1H, m).
13C H NMR spectroscopies (150MHz, CD3OD 17 carbon signals are provided in), low field area provides 10 sp2The carbon signal of hydridization, It is the carbonyl carbon signals of one one-tenth ester wherein at δ 174.6, it at a carboxyl carbon signal δ 131.8,136.9 is a double bond to be at δ 170.3 Carbon signal.It is a phenyl ring carbon signal at δ 116.2,116.2,122.9,133.1,133.1,163.5.Provided at high field region δ 65.2 One connects the carbon signal of oxygen, and δ 21.4,29.2,29.9 provides three methyl carbon signals.
In HMBC spectrums (Figure 20), there is phase in δ 1.09 (3H, s), 1.21 (3H, s) and δ 29.9,36.6,48.4,136.9 Close, illustrate that the two methyl are connected on δ 36.6 quaternary carbon.δ 1.72 (3H, s) and δ 41.6,131.8,136.9 has long-range phase Close, illustrate that this methyl is linked on δ 131.8 double key carbon.Related, the δ by δ 1.95 (1H, m) and δ 65.2,131.8,136.9 2.33 (1H, m) and δ 21.4,48.4,65.2,131.8,136.9 is related, δ 1.40 (1H, m) and δ 29.2,36.6,41.6,65.2 In the presence of long-range correlation, there is correlation, can draw structure fragment A in δ 1.74 (1H, m) and δ 36.6,65.2.By δ 6.84 (2H, dd, J =2.4,8.4Hz) related, δ 7.90 (2H, dd, J=2.4,8.4Hz) and δ 116.2 with δ 116.2,122.9,133.1 be present, 133.1,163.5,170.3 have correlation, can draw structure fragment B.In addition, from being understood to the carbonyl carbon signals of High-Field displacement, The carbonyl carbon is into ester.Therefore structure C can be drawn by A, B fragment.To sum up analyze, can determine that the compound structure for (4- hydroxyls- 2,6,6- trimethyls)-cyclohexenecarboxylic acid -4- hydroxyls-(benzoic acid) ester.
In CD is composed (see accompanying drawing 2), according to the spiral of unsaturated lactone rule, compound 2 presents negative at 233nm Cotton effects, it can determine whether that 4 are R types, this [1] consistent with compound picrocrocinic acid configuration, therefore determine should Compound is (4- hydroxyl -2,6,6- trimethyls)-cyclohexenecarboxylic acid -4- hydroxyls-(benzoic acid) ester.Retrieved through system documentation, be Have no the noval chemical compound of document report.
The compound 3 of table 41H-(600MHz in CD3OD),13C-NMR(150MHz in CD3) and HMBC data OD
Embodiment 6：The Identification of chemical structure of compound 4
White powder (methanol), it is dissolved in methanol, Molisch reacting positives.HR-ESI-TOF-MS spectrums provide accurate point of high-resolution Daughter ion peak m/z 521.2018 [M-H]-(Calcd.521.2023,-0.9ppm).With reference to its NMR data, its molecular formula is determined For C26H34O11, and calculate the compound and contain 10 degrees of unsaturation.
1H H NMR spectroscopies (400MHz, CD3OD in) low field area provide 1 group of 1,3,4- trisubstituted benzene ring proton signals δ 6.90 (1H, Br.s, H-2 "), 6.96 (1H, br.d, J=8.0Hz, H-5 "), 6.43 (1H, br.d, J=8.0Hz, H-6 "), 2 are trans double Key proton signal δ 6.41 (1H, d, J=16.0Hz, H-8 "), δ 7.63 (1H, d, J=16.0Hz, H-7 "), 1 alkene Hydrogen Proton letter Number δ 6.25 (1H, s, H-4).High field region provides 1 methoxy proton signal δ 3.87 (3H, s), 2 methyl proton signal δ 1.01 (3H, s), 1.11 (3H, s).By β-D-Glucose anomeric proton signal δ 4.44 (1H, d, J=8.0Hz, glc-1) and 6 protons Signal δ 4.32 (1H, br.d, J=11.6Hz), 4.51 (1H, br.d, J=11.6Hz) are understood, a Portugal in the molecular structure be present Grape glycan molecule, and 6 are acylated, and 1 is substituted.
13C H NMR spectroscopies (100MHz, CD3OD 26 carbon signals are provided in), low field area provides 12 sp2The carbon signal of hydridization, A carbonyl carbon signals are wherein shown as at δ 202.9.Carbon signal is glucose end group carbon signal at δ 104.7.It is a first at δ 57.0 Epoxide carbon signal, another high field region provide two methyl carbon signal δ 27.6,29.5.
(Figure 21) is understood by HMBC spectrums, δ 6.41 (1H, d, J=16.0Hz, H-8 ") and δ 125.7,147.5,169.1 is present Remotely related, there is long-range correlation in δ 7.63 (1H, d, J=16.0Hz, H-7 ") and δ 107.1,125.7,169.1, can draw one Phenylpropenoyl structure fragment.δ 4.32 (1H, br.d, J=11.6Hz, glc-6), 4.51 (1H, br.d, J=11.6Hz, glc- 6) it is long-range related to the presence of δ 169.1, illustrate that glucose sugar 6 is acylated, be connected with phenylpropenoyl.By δ 4.47,4.53 with There is correlation in δ 164.1,125.6, δ 3.83 and δ 36.4,50.7,164.1 has long-range correlation, δ 2.01,2.72 and δ 202.9, 36.4,50.7 is related, and δ 1.01,1.11 and δ 36.4,50.2,50.7 is related, can draw structure fragment A.δ 4.44 (1H, d, J= 8.0Hz, glc-1) long-range related, the deducibility glucose 1 and 10 methylol phases in said structure fragment to δ 72.2 be present Even.To sum up analyze, can draw the relative configuration of the compound for 6'-O- (3 "-methoxyl group -4 "-hydroxyl-phenylpropenoyl)-β - D-Glucose -1'-O-7- methyl-(6- methylol -1,1- dimethyleyelohexane -4- alkene -3- ketone) glycosides.
(accompanying drawing 4) is composed by CD to can be seen that, positive Cotton effects are presented at 235nm, 224nm and 207nm, are at 251nm Now negative Cotton effects.It is beta configuration to understand that C-7 connects Oxymethylene and the one of methene protons of C-2 (δ 2.72), so Judge the absolute configuration of the compound for R configurations[3].To sum up can conclude that the compound for (7R) -6 '-O- (3 "-methoxyl group -4 " - Hydroxyl-phenylpropenoyl)-β-- the O-7- of D-Glucose -1 ' methylols-(6- methylol -1,1- dimethyleyelohexane -4- alkene -3- ketone) Glycosides.
The compound 4 of table 51H-(400MHz in CD3OD),13C-NMR(100MHz in CD3) and HMBC data OD
Embodiment 7：The Identification of chemical structure of compound 5
Faint yellow indefinite form powder (methanol), is dissolved in methanol, Molisch reacting positives.HR-ESI-TOF-MS spectrums provide height Differentiate quasi-molecular ion peak m/z 345.1540 [M-H]-(Calcd.345.1549,1.2ppm).With reference to its NMR data, it is determined that Its molecular formula is C16H26O8, and calculate the compound and contain 4 degrees of unsaturation.
1H H NMR spectroscopies (400MHz, CD3OD 1, low field area alkene Hydrogen Proton signal δ 6.23 (1H, br.s, H-4) in).High field region Provide 2 methyl proton signal δ 1.01 (3H, s), 1.11 (3H, s).By β-D-Glucose anomeric proton signal δ 4.25 (1H, d, J=7.8Hz, glc-1) and 6 proton signal δ 3.67 (1H, dd, J=12.0,5.2Hz), 3.86 (1H, dd, J=12.0, 1.6Hz) understand, a glucose molecule in the molecular structure be present.
13C H NMR spectroscopies (100MHz, CD3OD 16 carbon signals are provided in), low field area provides 3 sp2The carbon signal of hydridization, A carbonyl carbon signals are wherein shown as at δ 202.9.Carbon signal is glucose end group carbon signal at δ 104.6.Another high field region provides Two methyl carbon signal δ 27.6,29.5.
(Figure 22) is understood by HMBC spectrums, related, δ 3.83 and δ 36.5 by δ 4.43,4.57 and δ 164.0,125.5 be present, 50.7,164.0 have long-range correlation, and δ 2.00,2.71 and δ 202.9,36.5,50.7 is related, δ 1.01,1.11 and δ 36.5,50.1, 50.7 is related, can draw structure fragment A.Long-range related, deducibility be present to δ 72.2 in δ 4.25 (1H, d, J=7.8Hz, glc-1) The glucose 1 is connected with 10 methylols in said structure fragment.To sum up analyze, the relative configuration that can draw the compound is 3- methylol -4- methylol -5,5- dimethyleyelohexane -2- ketenes-β-D-Glucose glycosides.
(accompanying drawing 3) is composed by CD to can be seen that, positive Cotton effects are presented at 233nm and 208nm, are presented what is born at 329nm Cotton effects.It is beta configuration to understand that C-7 connects Oxymethylene and the one of methene protons of C-2 (δ 2.71), so judging to be somebody's turn to do The absolute configuration of compound is R configurations.To sum up can conclude that the compound is (4R) -3- methylol -4- methylol -5,5- dimethyl Hexamethylene -2- ketenes-β-D-Glucose glycosides.Data above and document[2]Contrast, it is known that the compound and jasminoside B are each other Isomer.
The compound 5 of table 61H-(400MHz in CD3OD),13C-NMR(100MHz in CD3) and HMBC data OD
Embodiment 8：The Identification of chemical structure of compound 14
Faint yellow solid (methanol), HR-ESI-TOF-MS spectrums provide the [M+ of high-resolution quasi-molecular ion peak m/z 156.0796 H]+(Calcd.156.0786,2.3ppm).With reference to its NMR data, it is C to determine its molecular formula8H12O3, and calculate the chemical combination Thing contains 3 degrees of unsaturation.
1H NMR and13C NMR(600MHz,CD3OD a trans double bond signal δ is provided in) composingH7.29 (1H, t, J= 1.5Hz,H-3),δC130.8 (C-2), 151.2 (C-3), an aldehyde radical signal δH 9.75(1H,s,-CHO),δC 176.3(C- 1), a carbonyl carbon signals δC207 (C-6), two methyl carbon signal δH 2.18(3H,s,H-7),1.87(3H,s,2-CH3),δC 30.3(C-7),10.6(2-CH3), a mesomethylene carbon signal δH 2.87(2H,m,H-4),δC46.9 (C-4) and company's oxygen Tertiary carbon signal δH 5.32(1H,m,H-5),δC 78.9(C-5)。
HMBC spectrums (Figure 23) prompting, δH 1.87(3H,s,2-CH3) and C-1, C-2, C-3 correlation, H-3 and C-1, C-2, C-4 Correlation, H-4 and C-3, C-5, C-6 are related, and H-7 is related in W to C-4, and H-7 and C-5, C-6 are related.The compound structure can be drawn For (E) -5- hydroxy-2-methyls-hept-2-ene" -1,6- diketone.
The compound 14 of table 71H NMR(600MHz),13C NMR (150MHz) data
Embodiment 9：The Identification of chemical structure of compound 16
Yellow amorphous powder (methanol), it is dissolved in methanol, Molisch reacting positives.HR-ESI-TOF-MS spectrums provide accurate point Daughter ion peak m/z 711.2508 [M+H]+(Calcd.711.2500,1.8ppm).And combine its NMR data, thus it is speculated that its molecule Formula is C33H42O17, and calculate the compound and contain 13 degrees of unsaturation.
1H H NMR spectroscopies (400MHz, CD3OD) and13C H NMR spectroscopies (100MHz, CD3OD provided in) four double bond proton signals and Six double bond carbon signals, wherein including a trans double bond signal δH7.63 (1H, d, J=16.0Hz, H-7 " '), 6.44 (1H, d, J=16.0Hz, H-8 " '), δC147.4 (C-7 " '), 115.7 (C-8 " ').δH7.23 (2H, dd, J=2.0,8.0Hz, H- 2 " ', 6 " '), 6.80 (2H, dd, J=2.0,8.0Hz, H-3 " ', 5 " '), 3.87 (3H, s, 4 " '-OCH3) place's proton signal and δC 126.6 (C-1 " '), 159.6 (C-4 " '), 126.4 (C-2 " ', 6 " '), 115.3 (C-3 " ', 5 " '), 56.9 (4 " '-OCH3) place Contain a dibasic phenyl ring of 1,4- in carbon signal prompting structure.
In HMBC spectrums (Figure 24), H-7 " ' and C-1 " ', C-2 " ', 6 " ', C-9 " ' is related, 4 " '-OCH3To C-4 " ' related, H-8 " ' and C-9 " ', C-1 " ' is related, prompts a trans p- methoxybenzene acrylic acid groups be present.In addition, δH 4.40(1H,d, J=8.0Hz, H-1 "), 4.53 (1H, br.d, J=10.8Hz, H-6 "), 4.22 (1H, dd, J=2.0,10.8Hz, H-6 ") and δH4.71 (1H, d, J=8.0Hz, H-1 '), 4.09 (1H, dd, J=12.0,2.4Hz, H-6 '), 3.75 (1H, dd, J= 12.0,2.4Hz, H-6 ') two molecule glucoses be present in prompting structure, from anomeric proton coupling constant (8.0Hz), the Portugal Grape sugar is beta configuration.H-6 ' is related with C-6 ' to C-1 " and H-1 ", and the connection for prompting two glucose is 1 → 6.Remaining1H、13C NMR signal is similar to geniposide[8].H-1 is related to C-1 ' and H-1 ' with C-1, illustrates glucose 1 and 1 phase of Geniposide Even.It is related by H-6 " to C-9 " ', and knowable to the C-6 to low field displacement " signals, p- methoxybenzene acrylic is connected to another 6 of glucose.1H H NMR spectroscopies (400MHz, CD3OD) and13C H NMR spectroscopies data and trans-p- hydroxyl-cinnamyl groups of 6 "-O-- Geniposide-O-gentibioside contrast, it is basically identical[4], therefore judge the compound for trans-p- methoxyl group-propenyl benzenes of 6 "-O- Base-Geniposide-O-gentibioside.
The compound 16 of table 81H-(400MHz in CD3OD) and13C-NMR(100MHz in CD3OD) data
Embodiment 10：The Identification of chemical structure of compound 53
Yellow powder (methanol), ferric trichloride-potassium ferricyanide reacting positive, shows with the presence of phenolic hydroxyl group.UV(MeOH)λ max(logε):204 (3.82), 254 (3.79), 294 (2.93), HR-ESI-TOF-MS spectrums provide quasi-molecular ion peak m/z 369.1175[M+H]+(Calcd.369.1186, -2.6ppm), with reference to13C NMR、1H H NMR spectroscopies, thus it is speculated that its molecular formula is C17H20O9。
1H H NMR spectroscopies (400MHz, CD3OD low field area provides 2 aromatic signal δ 6.09 (1H, s, H-3), δ 6.24 in) (1H,s,H-6).High field region shows 1 methoxy proton signal δ 3.97 (3H, s, 7-OCH3), 1 methyl proton signal δ 2.40 (3H,s,2-CH3), another δ 4.39 (1H, d, J=7.6Hz, glc-1 ') place is glucose anomeric proton signal.13C H NMR spectroscopies (100MHz,CD3OD 17 carbon signals provided in), wherein low field area provide 9 sp2The carbon signal of hydridization, wherein at δ 184.5 For a carbonyl carbon signals.It is a methoxyl group carbon signal at high field region δ 56.8, is monomethyl carbon signal at δ 20.4, another δ 82.4, One group of Glucose Carbon signal to High-Field displacement is provided at 80.0,74.9,72.8,71.7,62.9, thus can determine that the glucose Carbon glycosides is formed with parent nucleus.In summary information will1H NMR,13C NMR datas and document[5]It is compared, it is seen that (the C- of δ 105.2 8) moved to low field, can determine whether that methoxyl group is connected in 7, therefore the deducibility compound is 7-methoxyl-5-hydroxy-2- Methylchromone-8-C- β-D-glucopyranoside, i.e. 7- methoxyl group-Isobiflorins.
The compound 53 of table 91H-(400MHz in CD3OD) and13C-NMR(100MHz in CD3OD) data
Embodiment 11：Pharmacodynamic study
Green Bamboo Leaf Liquor carries out the acute liver protective effect research of different type of impairments and to normal and immunity The Study immune regulation of low mouse.It protects the data of hepatic injury and Immune-enhancing effect as follows, illustrates to send out in Green Bamboo Leaf Liquor The active ingredient for waving protection hepatic injury and immunological enhancement is active ingredient of the prior art：Terpene, iridoids are yellow Ketone and flavonoid glycoside, chromogen ketone compounds, phenolic acid and phenolic glycoside class, aromatics, chinic acid class, Coumarins, lignanoids, steroid Body class, alkaloids, Furanones, furfural class, any type in long-chain fat acids compound.
Carbon tetrachloride causes mouse chemical damage protective effect research
The data such as following table of hepatic injury caused by it protects carbon tetrachloride：
1 couple of CCl4Cause the influence of hepatic injury mice organs coefficient
Compared with normal group, liver coefficient, Spleen coefficient and the Kidney coefficients of model group hepatic injury mouse have rise, but There was no significant difference.Compared with model group, liver coefficient, Spleen coefficient and the Kidney coefficients of Green Bamboo Leaf Liquor A-D groups have dropped It is low, but there was no significant difference.
The Green Bamboo Leaf Liquor mother liquor of table 10 is to CCl4Cause hepatic injury mouse liver coefficient, Spleen coefficient and Kidney coefficients influence (N=10)
Note：*Contrasted with model group, P ＜ 0.05；#Contrasted with blank group, P ＜ 0.05
The influence of 2 pairs of serum ASTs, ALT, NO
CCl4Model group is compared with normal group, and the horizontal significantly rise (P ＜ 0.01) of Serum ALT, AST, serum NO levels show Write rise (P ＜ 0.01).Show this experiment modeling success；Compared with model group, Green Bamboo Leaf Liquor mother liquor A-C groups are shown preferably Active (P ＜ 0.01, P ＜ 0.05), it is especially preferable with B groups.D groups serum AST, ALT levels have lowered compared with model group, but nothing Significant difference.Compared with model group, A-D group serum NO levels conspicuousness reduces (P ＜ 0.01, P ＜ 0.05), and F groups are without significantly Sex differernce.Result above shows Green Bamboo Leaf Liquor mother liquor to CCl4The protective effect of caused acute liver is probably base In terms of drop enzyme, anti-inflammatory.It the results are shown in Table 11.
The Green Bamboo Leaf Liquor mother liquor of table 11 is to CCl4Cause hepatic injury mice serum ALT, AST, NO influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01
T-AOC, GSH-PX influence in 3 pairs of hepatic tissues
CCl4For model group compared with normal group, hepatic tissue T-AOC, GSH-PX content significantly reduces (P ＜ 0.01), shows this Test modeling success；Compared with model group, Green Bamboo Leaf Liquor mother liquor A-D group hepatic tissue GSH-PX contents significantly raise (P ＜ 0.01, P ＜ 0.05)；Hepatic tissue T-AOC contents significantly raise (P ＜ 0.05) in Green Bamboo Leaf Liquor mother liquor B groups.Show Green Bamboo Leaf Liquor Mother liquor can improve CCl4Cause the oxidation resistance of hepatic injury mouse.It the results are shown in Table 12.
The Green Bamboo Leaf Liquor mother liquor of table 12 is to CCl4Cause hepatic injury murine liver tissue in T-AOC, GSH-PX influence (N= 10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01,#To be contrasted with blank group, P ＜ 0.05.
SOD, MDA influence in 4 pairs of hepatic tissues
CCl4For model group compared with normal group, hepatic tissue SOD activity significantly reduces (P ＜ 0.01), lipid peroxidation product MDA contents significantly raise (P ＜ 0.05), show this experiment modeling success.Compared with model group, Green Bamboo Leaf Liquor mother liquor A-D group livers Tissue SOD's activity significantly raises (P ＜ 0.01, P ＜ 0.05)；B, C groups lipid peroxidation product MDA contents are remarkably decreased (P ＜ 0.05).Show that Green Bamboo Leaf Liquor mother liquor can reduce CCl4The lipid peroxidation product of induced mice, there is certain protective effect. It the results are shown in Table 13.
Table 2-413 Green Bamboo Leaf Liquor mother liquors are to CCl4Cause hepatic injury murine liver tissue in SOD, MDA influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01,#To be contrasted with blank group, P ＜ 0.05.
The influence of 5 pairs of hepatic tissue pathology changes
From changes in histopathology, Normal group liver cell is arranged radially centered on central vein, knot Structure is normal；And CCl4Model group it is observed that serious Histopathologic change, such as：Centrilobular hepatic necrosis, macrophage, gas Ball sample deforms and substantial amounts of inflammatory cell infiltration；Compound Liver-benepitino Remedy group and green bamboo snake mother liquor A-D dosage groups can significantly improve this Hepatic tissue pathology changes, and as a result sees Fig. 6.
Thioacetamide causes mouse chemical damage protective effect research
1 Green Bamboo Leaf Liquor mother liquor causes the influence of hepatic injury mice organs coefficient to thioacetamide
Compared with normal group, the thymus gland coefficient conspicuousness of TAA model group hepatic injury mouse reduces (P ＜ 0.01), liver system Number has raised, but there was no significant difference, and there was no significant difference for Spleen coefficient, Kidney coefficients.Compared with model group, Green Bamboo Leaf Liquor The thymus gland coefficient and Spleen coefficient of each dosage groups of mother liquor A-D there are no significant difference, the liver coefficient of Green Bamboo Leaf Liquor mother solution C group show Work property reduces (P ＜ 0.05), and there was no significant difference for the Kidney coefficients of each dosage group.Positive drug legalon group compared with model group, Thymus gland coefficient, Spleen coefficient, liver coefficient, Kidney coefficients there are no significant difference.It the results are shown in Table 14.
The Green Bamboo Leaf Liquor mother liquor of table 14 causes hepatic injury mouse thymus coefficient, Spleen coefficient, liver coefficient, Kidney coefficients to TAA Influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
The influence of 2 pairs of serum ASTs, ALT, TBIL
TAA model groups are compared with normal group, Serum ALT, the horizontal significantly rise (P ＜ 0.01) of AST, TBIL.Show this examination Test modeling success；Compared with model group, the horizontal conspicuousnesses of Green Bamboo Leaf Liquor mother liquor B-D groups ALT reduce (P ＜ 0.01, P ＜ 0.05). The horizontal conspicuousness of each dosage groups AST, TBIL of A-D reduces (P ＜ 0.05, P ＜ 0.01).Show that Green Bamboo Leaf Liquor mother liquor is led to TAA The mouse liver injury of cause has certain protective effect.Positive drug legalon group Serum ALT, TBIL compared with model group is horizontal Conspicuousness reduces (P ＜ 0.05, P ＜ 0.01), the results are shown in Table 15.
The Green Bamboo Leaf Liquor mother liquor of table 15 TAA is caused hepatic injury mice serum ALT, AST, TBIL influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
GSH, T-AOC influence in 3 pairs of hepatic tissues
For TAA model groups compared with normal group, hepatic tissue GSH, T-AOC content significantly reduces (P ＜ 0.01), hepatic tissue SOD activity significantly reduces (P ＜ 0.01), and lipid peroxidation product MDA contents significantly raise (P ＜ 0.01), show that this experiment is made Mould success；Compared with model group, Green Bamboo Leaf Liquor mother liquor A-D group hepatic tissue GSH contents significantly raise (P ＜ 0.01, P ＜ 0.05), SOD activity significantly raises (P ＜ 0.01, P ＜ 0.05)；Each dosage group lipid peroxidation product MDA contents of B-D are notable Decline (P ＜ 0.01, P ＜ 0.05), T-AOC contents significantly raise (P ＜ 0.01, P ＜ 0.05).Show that Green Bamboo Leaf Liquor mother liquor can carry High TAA causes the oxidation resistance of hepatic injury mouse.It the results are shown in Table 16.
The Green Bamboo Leaf Liquor mother liquor of table 16 to TAA cause hepatic injury murine liver tissue in GSH, T-AOC, SOD, MDA influence ( N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
4 Histopathology results
Pathological tissue interpretation of result, Normal group：Liver cell is arranged radially centered on central vein, and liver is small Leaf, liver harden structure are normal, and without obvious cell infiltration, liver cell is without morphological changes such as denaturation, necrosis.Model control group：With Centered on central vein, there is piece focal necrosis in surrounding hepatic tissue, and average each high power field 14, location of necrosis has different journeys The cell infiltration of degree, large stretch of hepatic tissue liver cell enlargement, endochylema puffing, the expansion of part sinus hepaticus, there is a small amount of cell portal area Infiltration.Green Bamboo Leaf Liquor mother liquor A-D groups：Compared with model group, necrosis of liver tissue stove is still piece focal necrosis, but area diminishes, and is in Point, small pieces focal necrosis, location of necrosis have different degrees of cell infiltration, part hepatic tissue liver liver cell enlargement, and endochylema is dredged Song Hua, sinus hepaticus have no obvious expansion, and there is a small amount of cell infiltration portal area, and it is more obvious that B, D group improve lesion.Compound Liver-benepitino Remedy Group：There is spotty necrosis in hepatic tissue around the blood vessel of part, average each high power field 3, the infiltration of necrosis region visible inflammatory cell, quilt Hepatic tissue liver cell enlargement under film, endochylema puffing, sinus hepaticus have no obvious expansion, and there is a small amount of cell infiltration portal area, sees figure 7。
Ethanol causes Alcoholic Hepatic Injury protective effect research
1 animal behavior is observed
All mouse are before gavage alcohol, and equal normal diet drinking-water, fur gloss is submissive, and bouncing, body weight has difference Degree increase.After model group and each medicine group gavage alcohol, mostly astasia, walking is crooked, liquor-saturated soon sleeping, after a few houres from Dynamic wake up turns, but diet and amount of drinking water have all been reduced, and fur tarnishes, One's spirits are drooping depressed, and it is in black to defecate, foul smelling, Body weight loss.The animal state of medicine group is better than model group.
The influence of 2 pairs of alcohol induced Acute hepatic injury mice organs coefficients
Compared with normal group, the liver coefficient of model group significantly increases, and each medication therapy groups can significantly reduce liver Coefficient, and be in dose dependent, enlargement (P ＜ 0.01, the P ＜ that wherein Green Bamboo Leaf Liquor mother solution C, two groups of D can significantly reduce liver 0.05), thus it is speculated that medicine group can mitigate oedema and the hyperemia of liver.The Spleen coefficient of model group has certain compared to normal group Increase, prompt caused by Alcoholic that oedema and hyperemia may also occur in spleen in hepatic injury, cause quality to increase, each medicine Treatment group and DDB group can be in various degree reduction Spleen coefficient, but there is no significant difference.Specific data are shown in Table 17。
The Green Bamboo Leaf Liquor mother liquor of table 17 causes hepatic injury mouse thymus coefficient, Spleen coefficient, liver coefficient and kidney system to alcohol Several influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
The influence of 3 pairs of serum ASTs, ALT, TBIL
Alcohol model group is compared with normal group, Serum ALT, the horizontal significantly rise (P ＜ 0.01) of AST, TBIL.Show this examination Test modeling success；Compared with model group, Green Bamboo Leaf Liquor mother liquor A-D groups show preferably active (P ＜ 0.01, P ＜ 0.05), especially It is preferable with B groups.Show that rise of the Green Bamboo Leaf Liquor mother liquor to the acute liver serum transaminase caused by alcohol has one Fixed reduction effect.It the results are shown in Table 18.
The Green Bamboo Leaf Liquor mother liquor of table 18 alcohol is caused hepatic injury mice serum ALT, AST, TBIL influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01；##To be contrasted with blank group, P ＜ 0.01.
GSH, T-AOC, SOD, MDA influence in 4 pairs of hepatic tissues
For alcohol model group compared with normal group, hepatic tissue T-AOC, SOD, GSH content significantly reduces (P ＜ 0.01, P ＜ 0.05), lipid peroxidation product MDA content significantly raises (P ＜ 0.01), shows this experiment modeling success；With model group phase Than Green Bamboo Leaf Liquor mother liquor B, C group hepatic tissue GSH contents significantly raise (P ＜ 0.01, P ＜ 0.05)；C group hepatic tissues T-AOC contains Amount significantly rise (P ＜ 0.05)；Hepatic tissue SOD contents significantly raise (P ＜ 0.01, P ＜ in Green Bamboo Leaf Liquor mother liquor A-D groups 0.05)；A-D group lipid peroxidation product MDA contents significantly reduce, and show that Green Bamboo Leaf Liquor mother liquor can improve alcohol and cause hepatic injury small The oxidation resistance of mouse, reduce the content of lipid peroxidation product.It the results are shown in Table 19.
The Green Bamboo Leaf Liquor mother liquor of table 19 to alcohol cause hepatic injury murine liver tissue in GSH, T-AOC, SOD, MDA influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01,#To be contrasted with blank group, P ＜ 0.05.
The influence of 5 pairs of hepatic tissue pathology changes
From the point of view of histopathologic slide's result, Normal group lobuli hepatis structure is normal, no cell infiltration, liver cell with It is arranged radially centered on central vein, liver cell is without morphological changes such as denaturation, necrosis.Model control group central vein week Enclose hepatic tissue and piece focal necrosis occur, location of necrosis has different degrees of cell infiltration, large stretch of hepatic tissue liver cell enlargement, born of the same parents Puffing is starched, the expansion of part sinus hepaticus, there is a small amount of cellular infiltration portal area.Green Bamboo Leaf Liquor mother liquor A-D groups, compared with model group, liver Necrosis cooktop product reduces, and in point, small pieces focal necrosis, location of necrosis has different degrees of cell infiltration, part liver group Liver liver cell enlargement, endochylema puffing are knitted, sinus hepaticus has no obvious expansion, and there is a small amount of cell infiltration portal area, and C groups improve lesion It is more obvious.DDB positive controls, there is spotty necrosis, the leaching of necrosis region visible inflammatory cell in hepatic tissue around the blood vessel of part Moistening, hepatic tissue liver cell enlargement under envelope, endochylema puffing, sinus hepaticus has no obvious expansion, and there is a small amount of cell infiltration portal area, See Fig. 8.
The influence of 6 pairs of TNF-α expression
Experimental result shows, the expression without TNF-α in Normal group liver cell endochylema, model group murine liver tissue header Express and significantly increase in liver cell endochylema by area and sinus hepaticus.Each dosage Green Bamboo Leaf Liquor mother liquor group, which has, significantly reduces TNF-α in liver The effect of tissue expression, compared with model group, C, D group murine liver tissue TNF-α differential expression have significant statistical significance, See Fig. 9.
The influence of 7 couples of Fas and FasL expression
Research shows that the Apoptosis of Fas System-mediateds is one of important mechanisms of hepatopathy occurrence and development.Hepatopathy virus exists Surface of hepatocytes expression may one side cell cultured supernatant great expression Fas.On the other hand activation liver inner cell cytotoxic T cell CTL great expression FasL, both, which combine, causes Apoptosis.The typically no Fas/FasL of normal liver expression, or have few Measure Fas expression.Fas positive expressions are mainly expressed in endochylema, partly in after birth, and positive hepatocellular is primarily present in broken around leaflet Consider sample necrotic area to be worth doing, Fas expression is associated with course inflammatory activity degree.This research finds with immunohistochemical staining, Mice normal liver Dirty no Fas and FasL expression；Model group hepatic tissue Fas, FasL expression are substantially increased, and are expressed, are being drenched in extensive diffusivity It is obvious around Ba Xibaojinrunqu.Fas antigens are mainly expressed in liver cytoplasm, brown yellow granule, are diffused and irregular, liver is thin Also there is expression on cellular surface and sinusoidal endothelial cell surface：FasL expression is mainly based on endochylema, and minority is based on after birth, main collection In around central vein, while also have a large amount of distributions in lobuli hepatis.After the pretreatment of Green Bamboo Leaf Liquor mother liquor, murine liver tissue Damage mitigates, Fas and FasL antigen-positive cell numbers are reduced, and wherein C groups effect is the most obvious, relatively has with model group extremely aobvious Statistical significance (the P of work<0.01) Green Bamboo Leaf Liquor mother liquor, is prompted to reach CTL pairs of blocking by suppressing Fas/FasL expression The killing of liver cell, the hepatocellular apoptosis of alcohol induction is blocked, sees Figure 10, Figure 11.
The influence of 8 pairs of Bcl-2/Bax expression
Bcl-2 shows that Bcl-2 expresses in normal group height, model group low expression with Bax Immunohistochemical expression results, In contrast, in normal group low expression, model group is high to be expressed Bax.And Green Bamboo Leaf Liquor mother liquor and positive controls Bcl-2 expression compared with Model group strengthens, and Bax expression weakens compared with model group, prompts medicine to promote apoptosis by raising anti-apoptotic proteins Bcl-2 and lowering Expressing to play Anti-G value for protein Bax, is shown in Figure 12, Figure 13.
Acetaminophen causes mouse drug induced hepatic injury protective effect research
1 Abensanil causes the influence of hepatic injury mice organs coefficient
Compared with normal group, the thymus gland coefficient conspicuousness of AP model group hepatic injury mouse reduces (P ＜ 0.01), and hepatic injury is small The liver coefficient of mouse significantly raises (P ＜ 0.05), and Spleen coefficient conspicuousness rise (P ＜ 0.01), Kidney coefficients are poor without conspicuousness It is different.Compared with model group, the thymus gland coefficient of each dosage groups of Green Bamboo Leaf Liquor mother liquor A-D has raised, but without significant difference；Green bamboo snake The liver coefficient conspicuousness of distiller's yeast liquid B, C group reduces (P ＜ 0.05)；The equal conspicuousness of Spleen coefficient of Green Bamboo Leaf Liquor mother liquor A-D groups Reduce (P ＜ 0.01)；There was no significant difference for the Kidney coefficients of each dosage group.Positive drug DDB group is compared with model group, chest Gland coefficient has raised, but there was no significant difference, and Spleen coefficient conspicuousness reduces (P ＜ 0.01).It the results are shown in Table 20.
The Green Bamboo Leaf Liquor mother liquor Abensanil of table 20 causes hepatic injury mouse thymus coefficient, Spleen coefficient, liver coefficient and kidney Coefficient influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
The influence of 2 pairs of serum ASTs, ALT, TBIL
AP model groups are compared with normal group, Serum ALT, the horizontal significantly rise (P ＜ 0.01) of AST, TBIL.Show this experiment Modeling success；Compared with model group, the ALT levels of Green Bamboo Leaf Liquor mother liquor B, C dosage group significantly reduce (P ＜ 0.05, P ＜ 0.01), AST, TBIL level of each dosage groups of Green Bamboo Leaf Liquor mother liquor A-D significantly reduce (P ＜ 0.01), show green bamboo snake distiller's yeast Liquid has protective effect to the mouse drug induced hepatic injury caused by AP.Positive drug DDB group is compared with model group, serum AST, TBIL level significantly reduce (P ＜ 0.01), the results are shown in Table 21.
The Green Bamboo Leaf Liquor mother liquor of table 21 AP is caused hepatic injury mice serum ALT, AST, TBIL influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
GSH, T-AOC, SOD, MDA influence in 3 pairs of hepatic tissues
Compared with normal group, hepatic tissue GSH, T-AOC content significantly reduces (P ＜ 0.01) AP model groups, and hepatic tissue SOD lives Property significantly reduces (P ＜ 0.01), and lipid peroxidation product MDA contents significantly raise (P ＜ 0.01), show this experiment modeling into Work(.Compared with model group, each dosage group hepatic tissue GSH, T-AOC contents of Green Bamboo Leaf Liquor mother liquor B-D significantly rise (P ＜ 0.05, P ＜ 0.01), each dosage group hepatic tissue SOD activity of Green Bamboo Leaf Liquor mother liquor A-D significantly raises (P ＜ 0.01), lipid peroxidation production Thing MDA contents are remarkably decreased (P ＜ 0.05, P ＜ 0.01), show that Green Bamboo Leaf Liquor mother liquor can improve the antioxygen that AP causes hepatic injury mouse Change ability, caused lipid peroxidation product MDA in course of liver damage is removed, there is certain protection to make to drug induced hepatic injury With.Positive drug DDB group is compared with model group, and GSH, T-AOC content significantly raise (P ＜ 0.05, P ＜ in hepatic tissue 0.01), SOD activity significantly raises (P ＜ 0.01), and lipid peroxidation product MDA contents are remarkably decreased (P ＜ 0.01).As a result It is shown in Table 22.
The Green Bamboo Leaf Liquor mother liquor Abensanil of table 22 cause hepatic injury murine liver tissue in GSH, T-AOC, SOD, MDA influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
Con A cause immunological liver injury protective effect research
1 experimental animal general status
ICR mouse are successfully induced to establish acute hepatic injury model with Con A.Model group mouse tail vein injection Con A After (20mg/kg) 4h, autonomic activities significantly reduces, and no resistance is One's spirits are drooping, rolls up in cage, extraneous acousto-optic is stimulated ineffective It is quick, to look for food less, urine is yellow, and individual mice hair is upright, but without death in 8h.
2 couples of Con A cause the influence of hepatic injury mice organs coefficient
Compared with normal group, the thymus gland coefficient conspicuousness of Con A model group hepatic injury mouse reduces (P ＜ 0.01), spleen Coefficient, liver coefficient conspicuousness rise (P ＜ 0.01), there was no significant difference for Kidney coefficients.Compared with model group, green bamboo snake distiller's yeast Two groups of liquid B, C thymus gland coefficient conspicuousness rise (P ＜ 0.01), the equal conspicuousness drop of Spleen coefficient of Green Bamboo Leaf Liquor mother liquor A-D groups Low (P ＜ 0.01)；The equal conspicuousness of liver coefficient of each dosage groups of Green Bamboo Leaf Liquor mother liquor A-D reduces (P ＜ 0.01), each dosage group There was no significant difference for Kidney coefficients.Compared with model group, thymus gland coefficient has raised positive drug DDB group, but without conspicuousness Difference, Spleen coefficient conspicuousness reduce (P ＜ 0.01).It the results are shown in Table 23.
The Green Bamboo Leaf Liquor mother liquor of table 23 causes hepatic injury mouse thymus coefficient, Spleen coefficient, liver coefficient and kidney system to Con A Several influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01；##To be contrasted with blank group, P ＜ 0.01.
The influence of 3 pairs of serum ASTs, ALT, TBIL
Experimental result shows, tail vein injection ConA after 8 hours compared with normal group, Con A model groups Serum ALT, The horizontal significantly rise (P ＜ 0.01) of AST, TBIL, shows this experiment modeling success.Compared with model group, Green Bamboo Leaf Liquor mother liquor B, C Two groups of the horizontal conspicuousness reduction groups of ALT (P ＜ 0.05, P ＜ 0.01), there was no significant difference for the ALT levels that two groups of A, D；The leaf of bamboo The horizontal conspicuousnesses of AST, TBIL of blue or green distiller's yeast liquid A-D groups reduce (P ＜ 0.01), and result of the test shows Green Bamboo Leaf Liquor mother liquor pair Mouse liver injury caused by Con A has protective effect.Compared with model group, positive drug group serum AST, TBIL are horizontal significantly Reduce (P ＜ 0.05, P ＜ 0.01), the results are shown in Table 24.
The Green Bamboo Leaf Liquor mother liquor of table 24 Con A are caused hepatic injury mice serum ALT, AST, TBIL influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
GSH, T-AOC, SOD, MDA influence in 4 pairs of hepatic tissues
For Con A model groups compared with normal group, hepatic tissue GSH, T-AOC content significantly reduces (P ＜ 0.01), hepatic tissue SOD activity significantly reduces (P ＜ 0.01), and lipid peroxidation product MDA contents significantly raise (P ＜ 0.01), show this experiment mould Type success.Compared with model group, Green Bamboo Leaf Liquor mother liquor A-D group hepatic tissue GSH, T-AOC content significantly raise (P ＜ 0.05, P ＜ 0.01), SOD activity significantly raises (P ＜ 0.01)；Hepatic tissue lipid peroxidation product MDA contents are remarkably decreased (P ＜ 0.01), show that Green Bamboo Leaf Liquor mother liquor has certain protective effect to Con A induced mice immunological liver injuries.Positive drug group Compared with model group, GSH, T-AOC content significantly raise (P ＜ 0.01), and SOD activity significantly raises (P ＜ 0.01)；Lipid mistake Oxidation product MDA contents are remarkably decreased (P ＜ 0.01).It the results are shown in Table 25.
The Green Bamboo Leaf Liquor mother liquor of table 25 to Con A cause hepatic injury murine liver tissue in GSH, T-AOC, SOD, MDA influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01.
Immune-enhancing activity is studied
The influence of 1 pair of normal and hypoimmunity mouse weight, thymus gland coefficient and Spleen coefficient
Compared with normal group, body weight, thymus gland coefficient and the Spleen coefficient of model group mouse substantially reduce (P ＜ 0.01, P ＜ 0.05), especially thymus gland coefficient reduces significantly (P ＜ 0.01), prompts caused by cyclophosphamide hypoimmunity model modeling Success.Compared with normal group, normal mouse is after green bamboo snake mother liquor is given, body weight, thymus gland coefficient and the spleen system of each group mouse Number does not have significant difference.Compared with model group, the thymus gland coefficient of the hypoimmunity mouse of Green Bamboo Leaf Liquor A-F groups significantly rises High (P ＜ 0.01, P ＜ 0.05)；A, two groups of B Spleen coefficient significantly raises (P ＜ 0.05)；And the body weight of hypoimmunity mouse There was no significant difference compared with model group, the results are shown in Table 26.
The Green Bamboo Leaf Liquor mother liquor of table 26 to normal and hypoimmunity mouse weight, thymus gland coefficient and Spleen coefficient influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01,#To be contrasted with blank group, P ＜ 0.05.
2 pairs of normal and the mouse liver coefficient of hypoimmunity and influences of Kidney coefficients
Compared with normal group, the rise of model group mouse liver coefficient, but there was no significant difference；Kidney coefficients rise presents aobvious Write sex differernce (P ＜ 0.05).After normal mouse gives Green Bamboo Leaf Liquor mother liquor, its liver, Kidney coefficients are compared with normal group without aobvious Write sex differernce.Compared with model group, liver, the Kidney coefficients of hypoimmunity mouse significantly reduce (P ＜ 0.01, P ＜ 0.05) 27, be the results are shown in Table.
The Green Bamboo Leaf Liquor mother liquor of table 27 to normal and hypoimmunity mouse liver, Kidney coefficients influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；#To be contrasted with blank group, P ＜ 0.05.
3 couples of serum I FN- γ, IL-6 and LSZ influence
For model group compared with normal group, IL-6, IFN-γ content are remarkably decreased (P ＜ 0.01) in serum, illustrate this experiment Modeling success.Normal mouse is after the Green Bamboo Leaf Liquor mother liquor of A-F dosage groups is given, compared with normal group, in addition to E, F group, remaining Each dosage group blood serum IL-6, IFN-γ content have raised.Wherein, Green Bamboo Leaf Liquor mother liquor A, B groups IL-6 contents rise compared with For notable (P ＜ 0.01), there was no significant difference for remaining group.
In addition, the low mouse of endoxan immunogenicity is after the Green Bamboo Leaf Liquor mother liquor of A-F dosage groups is given, with model group Compare, each dosage group blood serum IL-6, IFN-γ content have raised.Wherein, A-D dosage groups blood serum IL-6, IFN-γ content With significant difference (P ＜ 0.01) compared with model group.It is normal and immune that data above illustrates that Green Bamboo Leaf Liquor mother liquor can strengthen The cell factor IL-6, IFN-γ of the low mouse of power secretion, so as to strengthen autoimmunity, the results are shown in Table 3-3.
In addition, serum lysozyme is to be synthesized by macrophage and can be promptly released into a kind of extracellular important hydrolase, It is a kind of important nospecific immunity factor of body defenses.This experimental evaluation ZYQL is to mice serum lysozyme activity Influence, the results are shown in Table 28, compared with normal group, mouse independent gavage Green Bamboo Leaf Liquor mother liquor 200mg/kg and 400mg/kg, serum Antalzyme activity has different degrees of raising, has significant difference (P<0.05, P<0.01) green bamboo snake distiller's yeast, is individually given During liquid 50mg/kg and 100mg/kg dosage, there was no significant difference for serum lysozyme level；After Cy is injected intraperitoneally, with normal control Group is compared, and serum lysozyme level significantly reduces (P<0.01)；Compared with Cy groups, each dosage group serum lysozymes of ZYQL+Cy are lived Power has different degrees of lifting (P<0.05, P<0.01), wherein 200mg/kg Green Bamboo Leaf Liquor mother liquor coordinates Cy effect pole Significantly (P<0.01).
The Green Bamboo Leaf Liquor mother liquor of table 28 is to IFN-γ in normal and hypoimmunity mice serum, the shadow of IL-6 and LSZ contents Ring (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01
SOD, GSH-PX, CAT influence in 4 pairs of spleen tissues
Model group is compared with normal group, and GSH-PX, CAT content and SOD activity, significantly reduce (P ＜ in spleen tissue 0.01) this experiment immunodeficiency models modeling success, is illustrated.Normal mouse awards the Green Bamboo Leaf Liquor mother liquor of various concentrations in gavage After (A-F groups), in spleen tissue, GSH-PX, CAT content and SOD activity significantly rise (P ＜ 0.01, P ＜ 0.05), illustrate bamboo Leaf green grass or young crops wine can improve normal mouse vivo oxidation activities of antioxidant enzymes, so as to strengthen mouse immunity of organisms.As shown in Table 29, bamboo Leaf green grass or young crops mother liquor B, C group is compared with other group positive effects.
In addition, Green Bamboo Leaf Liquor mother liquor can also improve GSH-PX, CAT in caused by cyclophosphamide hypoimmunity Mice Body The activity of content and SOD, compared with model group, there is significant difference (P ＜ 0.01, P ＜ 0.05), the results are shown in Table 29.
The Green Bamboo Leaf Liquor mother liquor of table 29 to SOD, GSH-PX, CAT in normal and hypoimmunity mouse boosting tissue influence (N=10)
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；##To be contrasted with blank group, P ＜ 0.01,#To be contrasted with blank group, P ＜ 0.05.
Influence (carbon particle clearance) of the 5 Green Bamboo Leaf Liquor mother liquors to mouse monokaryon mononuclear phagocyte system phagocytic function
After injecting Cy (100mg/kg), compared with Normal group, mouse phagocyte imdex significantly reduces Cy groups.With just Normal group compares, and individually gives 400mg/kg Green Bamboo Leaf Liquor mother liquor, and mouse phagocyte imdex can be made to raise (P<0.05).With mould Type group is compared, immunosupress caused by 100mg/kg, 200mg/kg, 400mg/kg Green Bamboo Leaf Liquor mother liquor can resist Cy, can Significantly improve phagocyte imdex (P<0.05, P<0.01) 30, be the results are shown in Table.
Influence (n=10) of the Green Bamboo Leaf Liquor mother liquor of table 30 to mouse monokaryon mononuclear phagocyte system phagocytic function
Note：**Contrasted with model group, P ＜ 0.01,*Contrasted with model group, P ＜ 0.05；#To be contrasted with blank group, P ＜ 0.05.
Embodiment 12：Bioactivity research
Any one compound carries out anti-inflammatory, cholinolytic ester in effective monomer chemical composition compound 1-78 in the present invention Enzyme, hepatocyte protective effect research.Result of study shows, it is thin that anti-inflammatory, anticholinesterase, Vitro hepatic are played in Green Bamboo Leaf Liquor The active ingredient of born of the same parents' protective effect is any one in compound 1-78 involved in the present invention.
Its anti-inflammatory, anticholinesterase, hepatocyte protective effect result such as following table 31：
Each monomeric compound extracorporeal anti-inflammatory of table 31, anticholinesterase and hepatocyte protection activity
aEach monomeric compound concentration is 50 μ Μ
bLiver protective effect positive controls
cAnti-inflammatory activity positive controls
dAnti- AChE active controls group
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Claims (7)

  1. A kind of 1. following compound of isolated structural formula from Green Bamboo Leaf Liquor：
  2. 2. application of the compound in liver protection, the medicine of Immune-enhancing effect, health products and food is prepared described in claim 1.
  3. 3. compound described in claim 1 prepare liver protection, Immune-enhancing effect functional food in application.
  4. 4. a kind of pharmaceutical composition, contain the compound described in claim 1.
  5. 5. a kind of health-care food composition, contain the compound described in claim 1.
  6. 6. the pharmaceutical composition described in claim 4, is prepared into pharmaceutical dosage forms as needed, it is selected from：Oral dosage form, Injection form, external preparation form, suppository form.
  7. 7. the health-care food composition described in claim 5, it is characterised in that the food compositions are selected from：Beverage, dairy products, Cake, candy.
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