CN1657090A - Extract of prinsepia wood and its bioactivity function - Google Patents

Extract of prinsepia wood and its bioactivity function Download PDF

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CN1657090A
CN1657090A CN 200410039587 CN200410039587A CN1657090A CN 1657090 A CN1657090 A CN 1657090A CN 200410039587 CN200410039587 CN 200410039587 CN 200410039587 A CN200410039587 A CN 200410039587A CN 1657090 A CN1657090 A CN 1657090A
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pue
pup
puo
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丁庆
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Abstract

An extract of prinsepia wood contains PUO, PUE and PUP. It can play the roles of regulating immunity, resisting against radiation and inflammation, increasing leucocytes and decreasing blood fat.

Description

Folium Prinsepiae Utilis extract and bioactive functions thereof
Folium Prinsepiae Utilis is a rosaceous plant, has another name called the Flos Mume thorn, and popular name Folium Prinsepiae Utilis, formal name used at school are Prinsepia utilis Royle. machaka, and be high 1 ~ 2 meter.Branch tool rib, celadon, normal adularescent Refined Mercurous chloride, tool scolus, long 8 ~ 20 millimeters.Single leaf alternate or grow thickly, ground paper matter is to keratin; Narrow avette to lanceolar, long 3 ~ 6.5 centimetres, wide 1 ~ 2.2 centimetre, blunt circle of base portion or wedge point, tip be point or short point gradually, edge tool serration, or several be full edge, the two sides does not have hair; Long 5 ~ 10 millimeters of petiole; Stipule is tiny, and the place is deposited or come off.The raceme axil is given birth to or is given birth to dried side shoot top, spends 3 ~ 8, white, sepal 5, subcircular; Petal 5 falls wealthy avette or oblate; The most multiple rows of stamen; Gynoecium 1, ovary is upper.The drupe ellipse, dark violet redness when ripe has Refined Mercurous chloride, and base portion has accrescent sepal.Be born in hillside or the trench two sides bushes and the depression, the roadside.Distribution Yunnan, Guizhou, Sichuan, ground such as Taiwan.The root of this plant (Radix prinsepiae utilis), fruit (Fructus prinsepiae utilis) is hyoscine also.
Hyperlipidemia is as atherosclerosis, one of important risk factor of cardiovascular and cerebrovascular disease.The invention reside in the spray (stem) of Folium Prinsepiae Utilis, leaf, fruit are extracted wherein effective active composition with modern means of science and technology respectively.After now extracting method and zoopery and embodiment being recited in.
1. the extracting method of the fruit of Folium Prinsepiae Utilis, stem, leaf:
1.1 the extraction of Folium Prinsepiae Utilis fruit: traditional extracting method is to carry out the machinery oil expression after the steaming and decocting, and not only yield is low, and oil presses endless, and residue is good utilisation not.Now use following solvent extraction, not only the high residue medium oil of yield has been carried to the greatest extent.
1.1.1 embodiment 1 gets 100g in apparatus,Soxhlet's after aquatic foods or dried Folium Prinsepiae Utilis are really pulverized, and adds 200ml petroleum ether (30-60) in receiving bottle and refluxes 2 hours in 70 ℃ of water-baths.After reclaiming petroleum ether, get Folium Prinsepiae Utilis oil (Prinsepia UtilisOil, abbreviation PUO) 28g, yield 28%.Residue fully utilizes after giving over to.(seeing accompanying drawing 1)
1.1.2 embodiment 2 replaces embodiment 1 PetroChina Company Limited.'s ether with normal hexane, other every operations are the same.(seeing accompanying drawing 2)
1.1.3 each constituents that contains among the PUO: see Table shown in the 1.2-1.
Table 1.2-1
Effective ingredient Phylloxanthin Beta-carotene The dihydroxycarotene fatty acid ester ??Vc ??Va ??Ve ??Vk Unsaturated fatty acid
Content % 0.95 1.94 3.1 ??0.75 ??0.81 ??0.97 ??0.69 ??90.83
1.1.4 embodiment 3 adds 80% ethanol with above-mentioned 72g marc A in receiving bottle, still refluxed 4 hours in apparatus,Soxhlet's, concentrating under reduced pressure becomes paste behind the recovery ethanol, dry, pulverize, pack behind 100 mesh sieves excessively, be called for short alcohol extraction Folium Prinsepiae Utilis (Prinsepia UtiliS Extract, abbreviation PUE).Residue B continues comprehensive utilization.(see accompanying drawing 3, effective ingredient sees Table 1.2-2)
Table 1.2-2
Figure A20041003958700041
Annotate: [1] village is to 1992 the 23rd volumes of equality Chinese herbal medicine the 3rd phase 122-124 page or leaf.
[2] village is to 1991 the 13rd volumes of equality Chinese patent medicine the 10th phase 10-11 page or leaf
1.1.5 embodiment 4 adds 6 liters of deionized waters backflows after 4 hours with the marc B among the 4kg embodiment 3, filter, it is 85% that filtrate transfers to alcohol concentration with dehydrated alcohol, white flocculent deposit occurs, refilters, drain, get pale powder, yield is 18.5%, crude polysaccharides (Prinsepia UtiliS Polysaccharides, abbreviation PUP), output: 0.74kg.(see accompanying drawing 4, composition sees Table 1.2-2)
1.2 the extracting method of Folium Prinsepiae Utilis stem (spray):
1.2.1 after embodiment 5 pulverizes scapus, get 4kg and extract, get PUE 0.048kg, yield 1.2% by embodiment 3 operations.Stem slag C continues comprehensive utilization (see accompanying drawing 3, effective ingredient sees Table 1.2-2)
1.2.2 embodiment 6 presses embodiment 4 operations with embodiment 5 stem slag C3.8kg and extracts, and gets PUP 0.2964kg, yield 7.8%.(see accompanying drawing 4, effective ingredient sees Table 1.2-2)
1.3 the extracting method of Folium Prinsepiae Utilis leaf:
1.3.1 embodiment 7 presses embodiment 3 operations with cured leaf 4kg and extracts, and gets PUE 0.06kg, yield 1.5%.Leaf slag D continues to extract (see accompanying drawing 3, composition sees Table 1.2-2)
1.3.2 embodiment 8 presses embodiment 5 operations with leaf slag D3.94kg and extracts, and gets PUP 0.453kg, yield 11.5%.(seeing accompanying drawing 4)
2, zoopery:
The toxicity test of PUO, PUE., PUP: the equal Department Of Medicine, Peking University of following animal origin
Get 150 of body weight 18-22g ICR mices, be divided into three groups, 50 every group, carry out LD50 according to a conventional method and measure (intraperitoneal administration), the result is 967.00 ± 143.15mg/kg..Get 4 of healthy dogs, body weight is about about 10kg, divides two groups of contrast and administrations, intravenous administration.The result shows: before and after the administration, Canis familiaris L. body weight, appetite, spirit and hepatic and renal function, hemogram are all normal.Main organs (samples such as the heart, liver,spleen,kidney, lung, stomach, intestinal, pancreas and adrenal gland) appearance not damaged, tissue slice there is no because drug-induced pathological changes and intoxicating phenomenon illustrate that PUO, PUE, PUP are nontoxic substantially.
3, the bioactive functions of PUO, PUE, PUP
3.1 PUO, PUE, PUP are to the facilitation of immunity
3.1.1 PUO, PUE, PUP are to the influence of immune organ weight
Get 44 of ICR mices about body weight 16g, divide two groups of contrast and administrations, male and female half and half.Administration group lumbar injection PUO, PUE, PUP, every 2mg * 7d, matched group is given the equivalent normal saline, and administration finishes the back dissects next day, with the eye socket depletion method mice is put to death.Taking-up spleen and thymus is also weighed, and the results are shown in Table 3.1-1
Table 3.1.1-1 is to the influence of mouse spleen and thymic weight
Figure A20041003958700051
*p<0.01, **p>0.05
By table 3.1.1-1 as seen, PUO, PUE, PUP can obviously increase mouse spleen weight, compare with matched group, and spleen index increases by 0.46 times, but not obvious to the influence of thymus.
3.1.2 PUO, PUE, PUP are to the influence of macrophage phagocytic function
Get 20 of body weight 18-22g ICR mices, observe PUO, PUE, PUP influence, the results are shown in Table 3.1.2-1 macrophage phagocytic function with the creep plate method
Table 3.1.2-1PUO, PUE, PUP are to the influence of peritoneal macrophage phagocytic function
Figure A20041003958700061
*p<0.01?? **p<0.001
By table 3.2-1 as seen, PUO, PUE, PUP can obviously strengthen the phagocytic function of mouse macrophage, compare with matched group, and phagocytic index has increased by 3.17 times, and engulfing percent increases by 1.57 times.
3.1.3 PUO, PUE, PUP are to the influence of humoral immunization
Get 40 of body weight 18-22gIRC mices, press literature method and measure PUO, PUE, PUP, the results are shown in Table 3.1.3-1 the influence that serum hemolysin produces.
The influence that table 3.1.3-1PUO, PUE, PUP produce hemolysin
The Mus number Route of administration Dosage mg/kg * d Half hemolysis value ± SD HC50 ± SD
Matched group ????10 ????Ip ????- ????16.35±3.81
PUO PUE PUP ????10 ????10 ????10 ????ip ????ip ????ip ????50×7 ????50×7 ????50×7 ????21.63±4.01 *????23.24±5.81 *????25.36±3.17 *
*p<0.01
By table 3.3-1 as seen, PUO, PUE, PUP have the effect that increases half hemolysis value, and compare, and half hemolysis value (HC50) increases by 0.56 times approximately.
3.1.4 the influence of PUO, PUE, the immunity of PUP pair cell
3.1.4.1 the experiment of E rosette: get 40 of centrifuge tubes, delivery tube adds 0.2ml (25 μ g/ml) PUO, PUE, PUP, and control tube adds 0.2ml Hank ' s liquid, injects provisional quality standard by the literature method thymosin.Observe PUO, PUE, PUP to the influence that the E rosette forms, the results are shown in Table 3.1.4-1.
The influence that table 3.1.4-1 PUO, PUE, PUP form the E rosette
Figure A20041003958700071
*p<0.01
By table 3.4-1 as seen, PUO, PUE, PUP can obviously increase the formation rate of E rosette, compare with matched group, and formation rate can increase by 0.80 times.
3.1.4.2 lymphocyte transformation experiment: get fresh normal person's whole blood (anticoagulant heparin) 0.2ml, add contain the 2ml culture fluid (include RPMI 1640, each 100 μ/ml of mycillin, adding calf serum, to make into concentration be 20%, uses NaHCO 3Accent pH7.4) in the test tube, dividing three groups is matched group (normal saline), PUO, PUE, each 50 μ g of PUP, 37 ℃ hatch 56h after every pipe add 5 μ ci3H-UR, continue after shaking up to hatch to 72h, with 49 type glass fibre filter disc harvestings, counting the results are shown in Table 3.1.4-2 with FJ-2105 liquid scintillation instrument rolling counters forward DPM.
The influence that table 3.1.4-2 PUO, PUE, PUP transform normal person's Whole blood lymphocyte
*P<0.01
By table 3.1.4-2 as seen, PUO, PUE, PUP group incorporation increase by 1.11 times with contrast, and comparative descriptions three all has the LT effect of promotion, there is no evident difference each other.
3.1.5 PUO, PUE, PUP are to the erythrocytic agglutination of rat
Get SD kind rat, heart puncturing extracting blood, anticoagulant heparin experimentizes by literature method, the results are shown in Table 3.1.5-1
Table 3.1.5-1 PUO, PUE, PUP are to the influence of big several erythroagglutinations
Figure A20041003958700081
*P<0.01
By table 3.1.5-1 as can be known, PUO, PUE, PUP have the erythrocytic effect of coagulation, illustrate that it has the effect of agglutinin, participates in cytophagous activation and biophylaxis.
4, PUO, PUE, PUP are to the reparation and the protective effect of body injury
4.1 PUO, PUE, PUP are right 60The radiating protective effect of Co gamma-rays
Symptoms such as get 50 of body weight 22gIRC mices, after the administration mice is shone, accumulated dose is 800 roentgens, shines after three days, and few food, few drink, loose stool appear in mice, the loose nothing of fur is damp and lose weight were observed one month, the results are shown in Table 4.1-1.
4.1-1PUO, PUE, PUP are right for table 60The radiating protective effect of Co gamma-rays
The Mus number Route of administration Dosage mg/kg * d Mean survival time (d) ± SD
Matched group ????25 ????ip ????- ????6.36±2.95
??PUO ??PUE ??PUP ????25 ????25 ????25 ????ip ????ip ????ip ????100×7 ????100×7 ????100×7 ????10.31±4.98 *????11.44±5.83 *????13.15±6.17 *
*P<0.01 (ip is a lumbar injection)
By table 4.1-1 as seen, PUO, PUE, PUP have the certain protection effect to radiation, compare with matched group, and mean survival time improves 0.80 times.
4.2 the effect of the leukopenia due to PUO, PUE, the PUP antagonism ring phosphamide
Get 40 of body weight 22gIRC mices, administration group lumbar injection PUO, PUE, PUP to the 7 days, injection ring phosphamide 2mg continued administration two days then in addition, checked leukocyte count routinely before and after the administration, the results are shown in Table 4.1-2.
The effect that table 4.1-2 PUO, PUE, PUP resist the leucocytes reduction that is caused by the ring phosphamide
*P<0.001
By table 4.1-2 as seen, PUO, PUE, PUP have the effect of obvious antagonism by the leukopenia that causes of ring phosphamide.
5, the antiinflammatory action of PUO, PUE, PUP
Get 10 of body weight 250-300gSD rat, press literature method, rat toes swelling method is tested.Ovum Gallus domesticus album solution with 0.1ml10% causes inflammation, the results are shown in Table 5-1.
Table 5-1 PUO, PUE, PUP are to the effect of anti-inflammatory
Figure A20041003958700092
P<0.01
By table 5-1 as seen, PUO, PUE, PUP promptly show preferably effect to anti-inflammatory causing scorching back 1h.
6, PUO, PUE, PUP mix the influence of serum albumin to the 3H-leucine
Get 40 of body weight 14-16gICR kind mices, administration group lumbar injection PUO, PUE, PUP100mg/kg * 4d, fasting 16h before mixing, every Mus lumbar injection 3H-leucine 400 μ ci/kg (Oura H (1972) Effectof radix ginsing extract on serum protein synthsis, Chem Pharm Bull 21:980-986.) the results are shown in Table 6-1 during experiment.
Table 6-1 PUO, PUE, PUP mix the proteinic influence of mice serum to the 3H-leucine
P<0.01
By table 6-1 as seen, PUO, PUE, PUP can promote the proteic biosynthesis of ICR mice serum, compare with matched group, and incorporation can on average increase by 65%.
7, PUO, PUE, PUP are to the effect of blood fat reducing
7.1 PUO, PUE, PUP are to the influence of normal mice serum cholesterol
Get male mice, random packet.Successive administration 4d, 4h gets blood after the last administration, surveys serum cholesterol.Matched group TG (mg/dl) is 111.73 ± 13.83, test group 150-200mg/kg4d, the TC value is respectively 107.71 ± 12.5 and 103.24 ± 15.55, and P>0.05 (n=10) shows that PUO, PUE, PUP do not have obvious influence to normal mice serum cholesterol level.
7.2 effect to the formation of prevention hypercholesterolemia
Get male mice, be divided into hypercholesterolemia test control group, normal control group and height dosed administration group at random.Behind the fasting 4h, administration group PO irritates stomach.Behind the 2h, the equal intravenous injection 75% egg yolk Emulsion 0.5ml of administration group and hypercholesterolemia group.The tail vein is got blood and is surveyed serum cholesterol content behind the 20h.Result's (seeing Table 7.1) shows, mice is PO, PUO, PUE, PUP100,150,200,100mg/kg. serum cholesterol content 15% and 23% (P<0.05) that descends respectively respectively
Table 7.1 PUO, PUE, PUP are to the influence of mice serum total cholesterol level (X ± S)
Group The Mus number Dosage mg/kg * kg ??Tc(mg/dl)
Matched group ????10 ?113.87±9.65
Positive controls ????10 ?579.46±81.95
PUO ????10 ????100 ?491.08±53.27 **
PUE ????10 ????150 ?446.67±101.25 **
PUP ????10 ????200 ?405.41±51.24 **
Compare with test control group P<0.05
7.3 the hyperlipidemia rats blood fat is influenced
Get male rat, be divided into normal control group, high fat matched group and height dosed administration group.High fat cholesterol feed 10d is fed for 3 groups in the back [3], the group of administration simultaneously successive administration 10d, normal control group and high fat matched group po equivalent normal saline.Behind the 10th day fasting 12h, heart puncturing extracting blood is measured serum total cholesterol (TC) with high ferro-sulfuric acid process; Normal heptane-isopropyl alcohol mixed liquor extraction method is measured serum triglycerides (TG); Heparin-manganese chloride turbidimetry is then decided serum low-density LP cholesterol (LDL-C); Sodium phosphotungstate-magnesium precipitate method is measured serum high-density LP cholesterol (HDL-C).Result's (seeing Table 7.2) shows, rat po30mg/kg PUO, PUE, PUP, and the hyperlipidemia rats serum TC, TG and LDL-C reduce by 29%, 18% and 22% respectively; HDL-C/TC improves 41%.
Table 7.2PUO, PUE, PUP are to the influence of mice TC, TG, LDL-C, HDL-C, HDL-C/TC (X ± S)
Group The Mus number Dosage mg/kg * d ?TC ?mg/dl ?TG ?mg/dl LDL-C mg/dl HDL-C mg/dl HDL-C/TC
Matched group ?8 ?82.5±13.5 ?115.8±19.5 274.0±18.2 44.8±5.4 0.54±0.03
Positive controls ?8 ?161.2±20.4 ?233.2±25.9 485.0±65.7 54.3±6.9 0.34±0.03
PUO ?8 ?30×10 ?130.7±19.8 * ?216.3±9.2 * 332.0±54.9 * 41.5±7.1 * 0.35±0.03 *
PUE ?8 ?50×10 ?114.2±13.4 ** ?191.32±34.8 ** 377.0±65.6 ** 53.4±8.8 ** 0.48±0.04 **
PUP ?8 ?70×10 ?105.1±11.5 ** ?185.14±36.1 ** 301.1±58.4 ** 40.6±6.9 ** 0.56±0.01 **
P>0.05 *P<0.05 *Compare with positive controls P<0.01

Claims (3)

1, Folium Prinsepiae Utilis extract of the present invention is divided into water miscible polysaccharide PUP, and the PUO of grease, the PUE of pure dissolubility all have purposes such as the immunity of adjusting, radioprotective, leukocyte increasing, anti-inflammatory, blood fat reducing;
2, according to claim 1, the present invention separates and extracting method for Folium Prinsepiae Utilis;
3, according to claim 1, the scope 30-200mg/kg of the dosage of taking.
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102600340A (en) * 2012-03-30 2012-07-25 王子夫 Traditional Chinese medical modification quick powder for treatment of cancers
CN102628002A (en) * 2012-03-31 2012-08-08 华健 Preparation process for prinsepia utilis royle grease
CN104027322A (en) * 2014-06-20 2014-09-10 成都乾坤动物药业有限公司 Microcapsules of prinsepia utilis royle polysaccharides and preparation method of microcapsules
CN105982979A (en) * 2015-01-30 2016-10-05 伽蓝(集团)股份有限公司 Tender leaf extract of prinsepia utilis as well as preparation method and applications of tender leaf extract
CN106305429A (en) * 2016-09-12 2017-01-11 北华大学 In-vitro rapid propagation technology for prinsepia sinensis
CN110055128A (en) * 2019-05-20 2019-07-26 贵州师范大学 A kind of extracting method of himalayan prinsepia leaf fruit oil

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102600340A (en) * 2012-03-30 2012-07-25 王子夫 Traditional Chinese medical modification quick powder for treatment of cancers
CN102628002A (en) * 2012-03-31 2012-08-08 华健 Preparation process for prinsepia utilis royle grease
CN104027322A (en) * 2014-06-20 2014-09-10 成都乾坤动物药业有限公司 Microcapsules of prinsepia utilis royle polysaccharides and preparation method of microcapsules
CN104027322B (en) * 2014-06-20 2016-06-08 成都乾坤动物药业有限公司 A kind of micro-capsule of Prinsepia utilis polysaccharide and its preparation method
CN105982979A (en) * 2015-01-30 2016-10-05 伽蓝(集团)股份有限公司 Tender leaf extract of prinsepia utilis as well as preparation method and applications of tender leaf extract
CN105982979B (en) * 2015-01-30 2020-06-09 伽蓝(集团)股份有限公司 Tender leaf extract of prinsepia utilis royle and preparation method and application thereof
CN106305429A (en) * 2016-09-12 2017-01-11 北华大学 In-vitro rapid propagation technology for prinsepia sinensis
CN106305429B (en) * 2016-09-12 2018-04-20 北华大学 A kind of rapid propagation in vitro method of Prinsepia sinensis
CN110055128A (en) * 2019-05-20 2019-07-26 贵州师范大学 A kind of extracting method of himalayan prinsepia leaf fruit oil

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