CN102552337A - Extract of lethariella cladonioides active site for reducing fat and protecting liver and application for extract - Google Patents
Extract of lethariella cladonioides active site for reducing fat and protecting liver and application for extract Download PDFInfo
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- lethariella cladonioides
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- 241001030098 Lethariella cladonioides Species 0.000 title claims abstract description 57
- 239000000284 extract Substances 0.000 title claims abstract description 40
- 210000004185 liver Anatomy 0.000 title abstract description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 24
- 238000002360 preparation method Methods 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims abstract description 11
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 9
- 108090000526 Papain Proteins 0.000 claims abstract description 6
- 239000004365 Protease Substances 0.000 claims abstract description 6
- 102000004142 Trypsin Human genes 0.000 claims abstract description 6
- 108090000631 Trypsin Proteins 0.000 claims abstract description 6
- 229940055729 papain Drugs 0.000 claims abstract description 6
- 235000019834 papain Nutrition 0.000 claims abstract description 6
- 239000012588 trypsin Substances 0.000 claims abstract description 6
- 208000010706 fatty liver disease Diseases 0.000 claims abstract description 4
- 239000002994 raw material Substances 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 48
- 238000000034 method Methods 0.000 claims description 11
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- 208000004930 Fatty Liver Diseases 0.000 claims description 3
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- 230000000694 effects Effects 0.000 abstract description 8
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- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 4
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- 238000010241 blood sampling Methods 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
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- 241001450685 Corallium japonicum Species 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
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- 235000005590 Larix decidua Nutrition 0.000 description 1
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- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- 241001310330 Parmeliaceae Species 0.000 description 1
- KNAHARQHSZJURB-UHFFFAOYSA-N Propylthiouracile Chemical compound CCCC1=CC(=O)NC(=S)N1 KNAHARQHSZJURB-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
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- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
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- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
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- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- VZOPRCCTKLAGPN-ZFJVMAEJSA-L potassium;sodium;(2r,3r)-2,3-dihydroxybutanedioate;tetrahydrate Chemical compound O.O.O.O.[Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O VZOPRCCTKLAGPN-ZFJVMAEJSA-L 0.000 description 1
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- NRHMKIHPTBHXPF-TUJRSCDTSA-M sodium cholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 NRHMKIHPTBHXPF-TUJRSCDTSA-M 0.000 description 1
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses an extract of a lethariella cladonioides active site for reducing fat and protecting liver, and belongs to the technical field of the preparation of natural medicines, and application of the extract to preparation of a medicine for preventing and treating nonalcoholic fatty liver diseases. The extract is prepared by the following steps of: performing water extraction on dry lethariella cladonioides coarse powder serving as a raw material, performing alcohol precipitation, hydrolyzing by using trypsin and papain, dialyzing, drying and the like. The effects of the extract of the lethariella cladonioides active site in the aspect of preventing and treating nonalcoholic fatty liver diseases is comparable to that of compound methionine and choline bitartrate tablets, so the extract serving as a medicine which can be used for preventing and treating fatty liver diseases has relatively wide development and application prospects.
Description
Technical field
The invention belongs to the natural drug preparing technical field, be specifically related to a kind of extract and application in the medicine of preparation control non-alcoholic fatty liver disease thereof of Lethariella cladonioides fat-reducing liver-protecting active site.
Background technology
Lethariella cladonioides (Lethariella cladoniodes) is a kind of lichen that lichens door Parmeliaceae spun gold belongs to.Shape such as Corallium Japonicum Kishinouye, the blister color is red, and bitter in the mouth raw meat band is sweet.Mainly be distributed in the plateau in thousands of meters in Tibet, Jiu Zhaigou, Sichuan, Yulong Xueshan, yak mountain, medicine mountain, Deqie, Yunnan, Lijing Yulong Xueshan, Mount Taibai, Shaanxi Province height above sea level, be grown on larch, the withered trunk of fir.Ethnic groups as wild vegetable and tea-drinking, discovers that Lethariella cladonioides has effects such as heat clearing away, antiinflammatory, resisting fatigue, radioprotective with it both at home and abroad.The inventor finds that under study for action the Lethariella cladonioides water extract has effect for reducing blood fat.
The research of at present relevant Lethariella cladonioides, domestic only on a small quantity about the Lethariella cladonioides pharmacognosy and chemical constituent such as researchs such as volatile ingredient, aminoacid group nutritional labeling.About the systematic study of systematic study, Lethariella cladonioides fat-reducing liver-protecting active effective part and the effective ingredient aspect of Lethariella cladonioides blood fat reducing the liver protecting pharmacological effect aspect does not also have bibliographical information.
Summary of the invention
The object of the present invention is to provide a kind of extract of Lethariella cladonioides fat-reducing liver-protecting active site.
The present invention also aims to provide the application of extract in the medicine of the fatty hepatitis of preparation control of above-mentioned Lethariella cladonioides active site.
A kind of extract of Lethariella cladonioides active site, this extract obtains according to the following steps extraction to be raw material with the dry coarse powder of Lethariella cladonioides:
(1) takes by weighing the dry coarse powder 100g of Lethariella cladonioides, add the water logging bubble 1-3h of 15-25 times of quality, heated and boiled 30-60min; Secondary decocts and adds water 3-8 times quality; 60-100 ℃ of condition refluxed 20-50min merges decocting liquid, filters; Filtrating concentrating under reduced pressure under 50-70 ℃ of condition is dry, obtains the Lethariella cladonioides water extract;
(2) get the Lethariella cladonioides water extract and add water 100-300mL, dissolving back adds ethanol to be made and contains pure mass percent and reach 60-85%, 4 ℃ of cold preservation 12-36h; Centrifuging and taking precipitate, oven dry obtain Lethariella cladonioides alcohol hypostasis, and Lethariella cladonioides alcohol hypostasis is added the 200mL water dissolution, use 0.16-0.50g trypsin, 0.08-0.30g papain enzymolysis successively, and dialysis back reuse Sevage method is removed Deproteinization, and oven dry promptly gets.
This extract extracts according to following steps and obtains:
(1) takes by weighing the dry coarse powder 100g of Lethariella cladonioides, add the water logging bubble 2h of 20 times of quality, heated and boiled 45min; Secondary decocts and adds 5 times of quality of water, and 80 ℃ of condition refluxed 30min merge decocting liquid; Filter, filtrating concentrating under reduced pressure under 60 ℃ of conditions is dry, obtains the Lethariella cladonioides water extract;
(2) get the Lethariella cladonioides water extract and add water 200mL, dissolving back adds ethanol to be made and contains pure mass percent and reach 80%, 4 ℃ of cold preservation 24h; Centrifuging and taking precipitate, oven dry obtain Lethariella cladonioides alcohol hypostasis, and Lethariella cladonioides alcohol hypostasis is added the 200mL water dissolution, use trypsin, the 0.08-0.30g papain enzymolysis of 0.16-0.50g successively, and dialysis back reuse Sevage method is removed Deproteinization, and oven dry promptly gets.
The extract component of above-mentioned Lethariella cladonioides active site is mainly saccharide, accounts for 65% of active site gross mass, is mainly rhamnose and galactose.
The application of the extract of above-mentioned Lethariella cladonioides active site in the lipotropic medicine of preparation.
Said fatty liver is a non-alcoholic fatty liver disease.
Beneficial effect of the present invention: the extract of Lethariella cladonioides active site of the present invention can significantly improve each item liver function index of fatty liver rat: reduce the liver index, reduce the content of TC, TG, LDL-C, ALT and AST in the serum.Effect and the DONGBAO GANTAI of the extract of Lethariella cladonioides active site of the present invention aspect control non-alcoholic liver is suitable, can be used for lipotropic medicine and has development prospect as a kind of.
Description of drawings
Fig. 1: liver tissues of rats pathological section (HE * 200);
Among the figure, A-normal group, B-model group, the low group of C-HTP, the high group of D-HTP, E-DONGBAO GANTAI group.
Fig. 2 is the total sugar canonical plotting.
Fig. 3 is the reducing sugar canonical plotting.
The specific embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is further specified.
The extraction of embodiment 1 Lethariella cladonioides active site (HTP)
Take by weighing the dry coarse powder 100g of Lethariella cladonioides, add the water logging bubble 2h of 20 times of quality, heated and boiled 45min; Secondary decocts and adds 5 times of quality of water, and 80 ℃ of condition refluxed 30min merge decocting liquid; Filter, filtrating concentrating under reduced pressure under 60 ℃ of conditions is dry, obtains the Lethariella cladonioides water extract;
Get the Lethariella cladonioides water extract and add water 200mL, dissolving back adds ethanol to be made and contains pure mass percent and reach 80%, 4 ℃ of cold preservation 24h; Centrifuging and taking precipitate, oven dry obtain Lethariella cladonioides alcohol hypostasis, and Lethariella cladonioides alcohol hypostasis is added the 200mL water dissolution, use trypsin, the 0.08-0.30g papain enzymolysis of 0.16-0.50g successively, and dialysis back reuse Sevage method is removed Deproteinization, and oven dry promptly gets.
The mensuration of embodiment 2 Lethariella cladonioides active site (HTP) sugar contents
Method: adopt the phenolsulfuric acid method to measure total sugar content, 3,5-dinitrosalicylic acid (DNS) method is measured content of reducing sugar, and both differences are HTP content.
Step:
1, need testing solution preparation
The accurate title, decided HTP 0.2g, puts in the 10mL volumetric flask, adds the water standardize solution, shakes up and promptly get the HTP need testing solution.
2, the preparation of reference substance solution
Take by weighing 105 ℃ of rhamnose reference substance 0.0320mg of drying to constant weight, put in the 50mL volumetric flask, be dissolved in water and standardize solution, promptly getting concentration is 0.64mgmL
-1The reference substance stock solution, precision is measured 1.563mL and is put in the 10mL volumetric flask, adds the water standardize solution, shakes up, and promptly gets 0.1mgmL
-1Reference substance solution.
3, the preparation of 6% phenol solution
Take by weighing phenol 200g, with aluminium flake 0.2g, sodium bicarbonate 0.1g, normal pressure oil bath distillation; Collect 182 ℃ of fractions, take by weighing this fraction 80g, place the 100mL volumetric flask, be dissolved in water and be diluted to scale; Shake up the back to brown bottle, be 80% phenol liquid, 4 ℃ of cold preservations are subsequent use.Precision is measured 1.875mL 80% phenol liquid, puts in the 25mL volumetric flask, adds the water standardize solution, promptly gets 6% phenol liquid, faces and uses new system.
4, the preparation of DNS reagent
Take by weighing 3,5-dinitrosalicylic acid 3.15g is dissolved in the sodium hydroxide solution that 131mL concentration is 2.0mol/L; This solution is joined 250mL to be contained in the hydrothermal solution (water temperature is less than 45 ℃) of 92.5g sodium potassium tartrate tetrahydrate; Add 2.5g again and heavily steam phenol and 2.5g sodium sulfite, stirring and dissolving is settled to 500mL after the cooling; Be stored in the brown bottle week back use.
5, measure the selection of wavelength
5.1 total HTP measures the selection of wavelength
The accurate 0.1mgmL that draws
-1Reference substance solution 1.0mL, distilled water, each 0.1mL of HTP need testing solution put respectively in the 10mL tool plug test tube, and adding distil water is to 2.0mL; Precision is measured the solution of 1.0mL 6% phenol, adds, and behind the mixing, precision is measured the 5.0mL concentrated sulphuric acid; Slowly add, shake up, leave standstill 5min after, put in the boiling water bath; Take out behind the 15min, to be cooled to room temperature, from each tool plug test tube, draw 300 μ L respectively, put in the ELISA Plate; With the corresponding reagent is blank, 230～999nm scope interscan absorption spectrum, and reference substance and test sample all have absorption maximum at the 480nm place, so confirm that maximum absorption wavelength is 480nm.
6, the reducing sugar test wavelength is selected
Accurate respectively distilled water, the 0.64mgmL of drawing
-1Reference substance solution, each 0.5mL of HTP need testing solution put in the 10mL tool plug test tube, add 1.5mL DNS reagent, behind the mixing, put in the boiling water bath, take out behind the 5min, and be to be cooled to room temperature, adds water 4mL, mixing.From each tool plug test tube, draw 300 μ L respectively, put in the ELISA Plate interscan of 230～999nm scope; Absorption curve tapers off, no maximum absorption band, and before 550nm; DNS reagent has interference to measuring the result, and in conjunction with documents and materials, final definite mensuration wavelength is 550nm.
7, the drafting of standard curve
7.1 total sugar standard curve
Accurate respectively 0.1mgmL-1 reference substance solution 0.2,0.4,0.6,0.8, the 1.0mL of drawing; Put in the 10mL tool plug test tube, adding distil water adds the phenol solution of 1.0mL 6% to 2mL, and mixing slowly adds the 5.0mL concentrated sulphuric acid; Shaking up, leave standstill 5min, put in the boiling water bath and take out behind the heating 15min, is blank with the corresponding reagent; Measuring the OD value in the 480nm place, is abscissa with the content of rhamnose, and optical density is a vertical coordinate, the drawing standard curve.The standard curve equation is OD=0.0514C-0.0146 (r=0.9992), in 2.5～12.5 μ gmL-1 scopes, good linear relationship is arranged, and the result is as shown in Figure 2.
7.2 reducing sugar standard curve
The accurate respectively 0.64mgmL-1 reference substance solution 0.1,0.2,0.3,0.4 of drawing; 0.5mL, put in the 10mL tool plug test tube, add water to 0.5mL, add 1.5mL DNS reagent; Mixing is put in the boiling water bath and is taken out behind the heating 5min, is blank with the corresponding reagent, measures the OD value in the 550nm place; Content with rhamnose is abscissa, and optical density is a vertical coordinate, the drawing standard curve.The standard curve equation is OD=0.0229C-0.086 (r=0.9999), at 8～40 μ gmL
-1Good linear relationship is arranged in the scope.Result such as Fig. 3.
8, precision experiment
The accurate 0.1mgmL that draws
-16 parts of reference substance solution, each 0.8mL presses total sugar standard curve METHOD FOR CONTINUOUS DETERMINATION 6 times, calculates total sugar content, and RSD is 1.98%, shows that instrument precision is good.
9, stability experiment
Precision is drawn HTP and is supplied examination solution 0.4mL, puts in the 10mL tool plug test tube, and method is every at a distance from 1 absorbance of 0.5h mensuration, continuous 13 times down to press " 7.1 " item.Calculating RSD is 1.54%, shows that in the total sugar content mensuration, sample solution is stable in 6h.
10, sample determination
10.1 total sugar content is measured
Get the HTP need testing solution, the accurate 0.4mL that draws measures total sugar content by condition under " 7.1 " item, records sugar content 86.9%.
10.2 content of reducing sugar is measured
Get the HTP need testing solution, the accurate 0.4mL that draws measures content of reducing sugar by condition under " 7.2 " item, records sugar content 4.28%.
10.3HTP saccharinity determining
By above-mentioned HTP total sugar content and content of reducing sugar, the sugar content of calculating HTP effective site is 82.6%.
Embodiment 3 Lethariella cladonioides active sites (HTP) are to the effect of rat non-alcoholic fatty liver disease
One, method
(1) foundation of non-alcoholic fatty liver rat model
Choosing 80 of cleaning level SD rat (180 ± 20) g, is male entirely; Continue to raise 6 weeks with high glucose and high fat feedstuff (normal feedstuff 72.7%, cholesterol 2%, yolk powder 5%, Adeps Sus domestica 10%, propylthiouracil 0.2%, sucrose 10%, sodium cholate 0.1%) every day, sets up non-alcoholic fatty liver (NAFLD) rat model.
(2) get 80 of NAFLD rat models and 20 of healthy rats, conventional raise (normal group) of healthy rat, 20 NAFLD rat model high glucose and high fat feedstuffs are raised (model group), irritate clothes 0.2g.kg for 20
-1.d
-1The extract (HTP low group) of Lethariella cladonioides active site, irritate clothes 0.6g.kg for 20
-1.d
-1The water extract (HTP high group) of Lethariella cladonioides active site, irritate clothes 1.2g.kg for 20
-1.d
-1DONGBAO GANTAI (DONGBAO GANTAI group), all rats were raised for 6 weeks continuously.
(3) getting the rat of respectively organizing of cultivating 3d and 42d weighs; That gets 6 weeks of feeding respectively organizes rat; Put to death with the femoral artery blood sampling; Take by weighing liver wet weights, adopt
and calculate the liver index.
(4) femoral artery blood sampling serum sample is pressed 3600rmin
-14 ℃ of centrifugal 10min; Separation of serum and homogenate supernatant, the content of TC (T-CHOL), TG (triglyceride), ALT (alanine aminotransferase), AST (aspartic acid aminotransferase), LDLC (low-density lipoprotein cholesterol), HDLC (HDL-C) in the mensuration serum.
(5) get each group and put to death liver tissues of rats 0.2g and add normal saline (1: 9) homogenate and get supernatant and be made into 10% liver tissue homogenate, adopt chemical colorimetry that the SOD in the rat liver tissues of rats (superoxide dismutase), MDA (malonaldehyde), TC and TG content are measured.
(6) cut 1 hepatic tissue from each middle part of organizing the right lobe of liver of putting to death rat, with the fixing back preparation of 10% formalin paraffin section observation pathological change.
The statistical procedures of above experimental data is used SAS9.1.3 software and is carried out statistical analysis; Measurement data is expression with
, and many groups data mean relatively adopts One-Way ANOVA check and t check.
Two, result
(1) each group rat body weight, the wet quality of liver and the exponential statistical result of liver are shown: wet quality of the liver of model group and liver index obviously raise, and the administration group can both reduce body weight; The administration group all can reduce the wet quality of liver with the liver index and high group of HTP more obviously reduced liver index (table 1).Effect and the DONGBAO GANTAI of proof Lethariella cladonioides active site aspect the liver function that improves the hepatitis rat is suitable, and the HTP effect of high dose also is superior to DONGBAO GANTAI.
Table 1: each organizes rat body constitution amount, liver index variation
**P＜0.01，
#P＜0.05，
##P＜0.01.
(2) measurement result to TC, TG in the content of Serum TC, TG, ALT, AST, LDLC, HDLC and the hepatic tissue shows: raise with middle TG, the ALT of the serum of compared with normal model group; And the TC in TC, LDLC and the hepatic tissue, TG significantly raise in the serum; HDLC descends, and AST changes not obvious; The administration group can both reduce the TC in serum TC, TG, LDLC, ALT and the hepatic tissue; High group of group of HTP significantly reduces TC, TG, and the low group of HTP can significantly reduce LDLC; The model group HDLC that significantly raises; The low group of HTP significantly reduces ALT (table 2).Show HTP obviously blood fat reducing and liver fat and have the effect of protection hepatic injury.
Table 2 is respectively organized the content
of Serum TC, TG, HDL-C, LDL-C, ALT, AST and hepatic tissue TC, TG
*P＜0.05，
**P＜0.01，
#P＜0.05，
##P＜0.01
(3) the content measurement result to liver tissues of rats SOD, MDA shows, the SOD of model group descends and MDA obviously raises, and the administration group can both reduce MDA, and the low group of HTP more obviously reduces MDA.But HTP hangs down group, HTP high group can increased SOD.Explain that HTP can improve the body anti-oxidant vigor and suppress lipid peroxidation.
Table 3: to the influence
of liver tissues of rats SOD, MDA
*P＜0.05，
**P＜0.01，
#P＜0.05，
##P＜0.01.
(4) the hepatic tissue paraffin section Electronic Speculum figure of each group execution rat shows; The hepatic tissue of model group rat is the severe steatosis; The hepatocyte anthorisma; Nucleus is extruded the low group group of crushed element disappearance HTP, and high group of group of HTP and DONGBAO GANTAI group denaturation degrees all alleviate, and hepatocyte anthorisma symptom also alleviates (Fig. 1).
Claims (4)
1. the extract of a Lethariella cladonioides active site is characterized in that, this extract is a raw material with the dry coarse powder of Lethariella cladonioides, extracts according to following steps to obtain:
(1) takes by weighing the dry coarse powder 100g of Lethariella cladonioides, add the water logging bubble 1-3h of 15-25 times of quality, heated and boiled 30-60min; Secondary decocts and adds water 3-8 times quality; 60-100 ℃ of condition refluxed 20-50min merges decocting liquid, filters; Filtrating concentrating under reduced pressure under 50-70 ℃ of condition is dry, obtains the Lethariella cladonioides water extract;
(2) get the Lethariella cladonioides water extract and add water 100-300mL, dissolving back adds ethanol to be made and contains pure mass percent and reach 60-85%, 4 ℃ of cold preservation 12-36h; Centrifuging and taking precipitate, oven dry obtain Lethariella cladonioides alcohol hypostasis, and Lethariella cladonioides alcohol hypostasis is added the 200mL water dissolution, use 0.16-0.50g trypsin, 0.08-0.30g papain enzymolysis successively, and dialysis back reuse Sevage method is removed Deproteinization, and oven dry promptly gets.
2. according to the extract of the said a kind of Lethariella cladonioides active site of claim 1, it is characterized in that this extract extracts according to following steps and obtains:
(1) takes by weighing the dry coarse powder 100g of Lethariella cladonioides, add the water logging bubble 2h of 20 times of quality, heated and boiled 45min; Secondary decocts and adds 5 times of quality of water, and 80 ℃ of condition refluxed 30min merge decocting liquid; Filter, filtrating concentrating under reduced pressure under 60 ℃ of conditions is dry, obtains the Lethariella cladonioides water extract;
(2) get the Lethariella cladonioides water extract and add water 200mL, dissolving back adds ethanol to be made and contains pure mass percent and reach 80%, 4 ℃ of cold preservation 24h; Centrifuging and taking precipitate, oven dry obtain Lethariella cladonioides alcohol hypostasis, and Lethariella cladonioides alcohol hypostasis is added the 200mL water dissolution, use trypsin, the 0.08-0.30g papain enzymolysis of 0.16-0.50g successively, and dialysis back reuse Sevage method is removed Deproteinization, and oven dry promptly gets.
3. the application of the extract of the said Lethariella cladonioides active site of claim 1 in the lipotropic medicine of preparation.
4. according to the application of extract in the lipotropic medicine of preparation of the said Lethariella cladonioides active site of claim 3, it is characterized in that said fatty liver is a non-alcoholic fatty liver disease.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103947805A (en) * | 2014-05-04 | 2014-07-30 | 李滋星 | Lethariella cladonioides biological drink and preparation method thereof |
| CN109864911A (en) * | 2017-12-01 | 2019-06-11 | 伽蓝（集团）股份有限公司 | A kind of application for avenging tea extraction |
-
2012
- 2012-02-21 CN CN2012100426503A patent/CN102552337A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 崔璨等: "高血脂症性脂肪肝的研究近况", 《现代中西医结合杂志》 * |
| 庄筱葳: "西藏红雪茶多糖含量测定", 《中国实验方剂学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103947805A (en) * | 2014-05-04 | 2014-07-30 | 李滋星 | Lethariella cladonioides biological drink and preparation method thereof |
| CN109864911A (en) * | 2017-12-01 | 2019-06-11 | 伽蓝（集团）股份有限公司 | A kind of application for avenging tea extraction |
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