CN105287685A - Application of alcohol extract of viburnum sargentii koehne to preparation of medicines for treating atherosclerosis diseases - Google Patents
Application of alcohol extract of viburnum sargentii koehne to preparation of medicines for treating atherosclerosis diseases Download PDFInfo
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- CN105287685A CN105287685A CN201510482073.3A CN201510482073A CN105287685A CN 105287685 A CN105287685 A CN 105287685A CN 201510482073 A CN201510482073 A CN 201510482073A CN 105287685 A CN105287685 A CN 105287685A
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Abstract
The invention discloses application of an alcohol extract of viburnum sargentii koehne to preparation of medicines for treating AS (atherosclerosis) diseases. By taking C57BL/6J mice as blank control and taking a simvastatin medicated group as positive control, observation of influences of the viburnum sargentii koehne on AS and exploration of corresponding mechanisms are carried out after a high fat diet-induced apoE<-/-> mouse AS model is intragastrically given with the alcohol extract of the viburnum sargentii koehne for 12 weeks. According to results, the alcohol extract of the viburnum sargentii koehne evidently lowers the level of lipids in serum, inhibits oxidative stress and inflammatory reaction, decelerates formation of atheromatous plaques and lipid accumulation on arterial walls of the high fat diet-induced apoE<-/-> mouse and inhibits AS occurrence and development. By application of the alcohol extract of the viburnum sargentii koehne to preparation of the medicines for treating the AS diseases, not only is a novel usage of the alcohol extract of the viburnum sargentii koehne is developed, but also a novel approach of AS treatment is provided.
Description
Technical field
The present invention relates to Tianmu lake application, be specially the application of a kind of Tianmu lake ethanol extract in preparation treatment atherosclerosis medicine.
Background technology
Atherosclerosis (atherosclerosis, AS) is commonly encountered diseases and the frequently-occurring disease of serious harm human health, and the formation of vulnerable plaque is the main cause causing AS pathological changes late phase clinical acute symptom with breaking.Massive epidemiology investigation and experimentation show, hyperlipidemia, smoking, diabetes, hypertension, sex difference etc. are all that relevant risk factor occurs with AS, Richard etc. think that hyperlipemia (Hyperlipidemia, HLP) causes the immediate cause of AS and primary risk factor.Wherein serum cholesterol being especially proportionate of low-density lipoprotein cholesterol (LDL-C) level and AS.The research such as Berliner finds that LDL-C can change OxLDL ELISA (ox-LDL-C) into through the lipid peroxidation of free radical initiation in blood plasma or under endarterium, ox-LDL-C is the key component of endothelial injury, also be the potential inducer of the endothelial inflammation factor simultaneously, start atherosclerotic lesions.Human vascular endothelial plays a crucial role in AS evolution, and anti-inflammatory drug can regulate the function of blood vessel endothelium, and shows certain anti-AS pharmacological action.In AS process, macrophage apoptosis not only directly causes the smooth muscle cell of apoptosis and macrophage effectively not to be engulfed, promote formation and the increase of lipid core, and the apoptosis macrophage being rich in free cholesterol damages fibrous cap by release matrix degrading proteinase, and produce the inflammatory factor such as tumor necrosis factor-alpha, Interleukin-1β, cause secondary inflammation to react and necrosis, thus promote that speckle is unstable.
Therefore, restriction cholesterol absorption, reduces NHDL (non-HDL), high density lipoprotein increasing (highdensitylipoprotein, HDL), and inhibited oxidation stress level and arterial wall inflammation have become the important means of control AS.The adjustment of endothelial cell inflammation reaction may as the target spot finding Antiatherosclerosis medicine research.Therefore studying prophylactico-therapeutic measures and finding effective medicine is the research field that the whole world is very paid attention to, Chinese medicine has the advantage of uniqueness in hyperlipemia and cardiovascular and cerebrovascular disease preventing and treating, because of its curative effect affirmative, lasting, side effect is little and become first-selected medicine.
Tianmu lake (Viburnum.sargentiikoehne) also claims Folium et Cacumen Viburni Sargentii, chicken mock Caulis Viburni Dilatati and mountain silk ball etc., for Caprifoliaceae (Caprifoliaceae) Viburnum (ViburnumL.) machaka, be born in the ground such as hillside, shrubbery and roadside, originate in the ground such as northeast, North China.Fruit and branch and leaf all can be used as medicine, and are mainly used in anti-tic, spasmolytic, calmness and excited uterus.The many formula about Tianmu lake of China's folklore, are used for the treatment of cough, skin ulcer scabies and lumbago and skelalgia etc., therefore can excavate its medical value energetically.Research confirms, Tianmu lake is rich in plurality of active ingredients, and fruits nutrition enriches, and flavone and terpene substances, as its main component, have significant antioxidation and antiinflammatory action.
Summary of the invention
The object of the invention is to lack efficient safe drugs again for current atherosclerosis, the novelty teabag of Chinese medicine Tianmu lake ethanol extract in preparation treatment atheromatosis medicine is provided.
For achieving the above object, the technical scheme that the present invention takes is:
The application of Tianmu lake ethanol extract in preparation treatment atheromatosis medicine.
The application of Tianmu lake ethanol extract in preparation treatment atheromatosis medicine, the extracting method of described ethanol extract is: the extracting method of described ethanol extract is: dried by Tianmu lake fruit room temperature, then 40 order coarse powder are ground to form, get 500g powder in the ethanol of 95% and extract, the quality of described powder and the volume ratio of ethanol are 1:10, ethanol extract is condensed into extractum after filtering, every gram of extractum is equivalent to 3.49g crude drug, then by extractum with 5% sodium carboxymethyl cellulose be mixed with 2.5g crude drug/ml respectively, 5.0g crude drug/ml, 10.0g crude drug/ml, 4 DEG C save backup.
The application of Tianmu lake ethanol extract in the exogenous atherosclerosis medicine of preparation treatment
Described atheromatosis refers to the atherosclerosis that the factors such as hyperlipemia, oxidative stress or inflammation that high fat diet is induced cause.
This invention exploits a kind of novel drugs of preventing and treating atheromatosis: Tianmu lake ethanol extract, the apoE-that the present invention adopts high fat diet to induce/-Mus AS model, using simvastatin as positive control drug, gavage gave Tianmu lake ethanol extract after 12 weeks, observe the impact on blood fat and AS speckle, experimental result finds, Tianmu lake ethanol extract has obvious effect to control atheromatosis;
The present invention has also found the novelty teabag of Tianmu lake ethanol extract: treatment atheromatosis, has excavated the biological activity of Tianmu lake further, has expanded the range of application of this Chinese medicine;
Tianmu lake ethanol extract preparation method is simple, and cost is lower, and toxic and side effects is little, has the advantage of good treatment atheromatosis.
Accompanying drawing explanation
Fig. 1 Tianmu lake ethanol extract is to apoE
-/-the impact of lipid of mice.Tianmu lake ethanol extract significantly reduces apoE
-/-t-CHOL (TC) and low density lipoprotein, LDL (LDL-C) level in mice plasma, remarkable high density lipoprotein increasing (HDL-C) level, triglyceride (TG) horizontal there was no significant difference; (mean ± SD, n=8).
* p<0.01
vscD group;
# p<0.05, ## p<0.01vshFD group; CD, blank group; HFD, hyperlipidemia model group; SV, simvastatin group; TQ-L, Tianmu lake low dose group; TQ-M, dosage group in Tianmu lake; TQ-H, Tianmu lake high dose group;
Fig. 2 Tianmu lake significantly alleviates apoE
-/
-interleukin-6 (IL-6) in mice plasma, tumor necrosis factor (TNF-α), the expression of the inflammatory factors such as C reactive protein (CRP); (mean ± SD, n=8);
* p<0.01
vscD group;
# p<0.05, ## p<0.01vshFD group; CD, blank group; HFD, hyperlipidemia model group; SV, simvastatin group; TQ-L, Tianmu lake low dose group; TQ-M, dosage group in Tianmu lake; TQ-H, Tianmu lake high dose group;
Fig. 3 is at the apoE of high fat feed
-/-in mice, Tianmu lake ethanol extract significantly suppress the formation of arterial wall lipidosis and atheromatous plaque, alleviates arterial wall damage (n=8); Give Tianmu lake ethanol extract after 12 weeks, by the degree of impairment of oil red O stain display arterial wall; CD, blank group; HFD, hyperlipidemia model group; SV, simvastatin group; TQ-L, Tianmu lake low dose group; TQ-M, dosage group in Tianmu lake; TQ-H, Tianmu lake high dose group; A, aorta total length; B, base of heart; C, plaque area.
Detailed description of the invention
one, materials and methods
(1) preparation of Tianmu lake ethanol extract
Tianmu lake Fruit, from Mount Taishan, is identified by Taishan Hospital Chinese materia medica Senior Expert professor Li Tongde.Grind to form 40 order coarse powder after Tianmu lake fruit room temperature dries, get 500g powder and extract in the ethanol (M:V=1:10) of 95%.Ethanol extract filter after be condensed into extractum (3.49g crude drug/g), then by extractum with 5% sodium carboxymethyl cellulose be mixed with 2.5g crude drug/ml, 5.0g crude drug/ml, 10.0g crude drug/ml respectively, 4 DEG C save backup.
(2) animal grouping and process
40 male apoE
-/-purchased from Beijing HFK Bio-Technology Co., Ltd. [credit number: SCYK (capital) 2014-0004], adaptability is divided into 5 groups after raising 1 week to mice (20 ± 2g, 7 week age) at random:
Model group (high fat diet: 10% Adeps Sus domestica+10% yolk powder+0.30% cholesterol+normal diet);
Tianmu lake low dose group (high fat diet+2.5g crude drug/kg/d);
Dosage group (high fat diet+5g crude drug/kg/d) in Tianmu lake;
Tianmu lake high dose group (high fat diet+10g crude drug/kg/d);
Simvastatin group (high fat diet+simvastatin 20mg/kg/d), often organizes 8 animals.
Separately get 8 C57BL/6J male mices as blank group (full diet);
Tianmu lake and simvastatin are all dissolved in the sodium carboxymethyl cellulose of 5%, and its final concentration is respectively Tianmu lake 2.5g crude drug/ml, (5.0g crude drug/ml, 10.0g crude drug/ml and simvastatin 20.0g crude drug/ml, mouse stomach 12 weeks.
At 12 weekends, animal fasting 12 hours, takes Mouse Weight, and after urethane anesthesia fiber crops, endocanthion gets blood, gets serum for subsequent use after centrifugal.Weigh after getting the cleaning of liver physiology saline, base of heart is also with OCT embedding at once after the cleaning of ice normal saline, and be put in 80 DEG C and save backup, get aorta total length, 80 DEG C of preservations, for oil red O stain.
(3) Mouse Weight and liver weight analysis
Medication is after 12 weeks, takes mice and to weigh in regulating liver-QI weight, analyze the impact that Tianmu lake is heavy on high fat diet Mouse Weight regulating liver-QI.
(4) Analysis of blood lipid
Mice medication is after 12 weeks, get mice serum, according to description (Zhong Shengbeikong bio tech ltd), enzyme process (Biosino) measures total plasma cholesterol (TC), HDL-C (HDL-C), low density lipoprotein, LDL (LDL-C) and triglyceride (TG) content, observes Tianmu lake to the impact of lipid of mice.
(5) Tianmu lake is on the impact of oxidative stress level
Oxidative stress is the key factor that development occurs atherosclerosis, and its level is by lipid peroxidation product malonaldehyde (MDA) reflection, and the toxic action of active oxygen can by antioxidant as superoxide dismutase (SOD) antagonism simultaneously.The active reaction of plasma SOD, the MDA extent of reaction of oxidative stress.Get mice serum, use microplate reader to detect SOD and MDA according to reagent description (Science and Technology Ltd. is built up in Nanjing) active.
(6) inflammatory factor detects
Mice medication is after 12 weeks, get blood, according to description (Sai Mo flies bio tech ltd), Elisa method is adopted to detect inflammatory factor interleukin-6 (IL-6) in serum, the tumor promotion factor (TNF-α), the expression of C reactive protein (CRP) content.
(7) artery plaque deposition conditions is analyzed
In order to detect the impact of Tianmu lake on atheromatous plaque, get apoE
-/-mouse aorta total length, adopts oil red O stain technology, detects arterial wall deposition conditions, dirty basal slices of simultaneously coring, oil red O stain, observes Aortic injury situation.
(8) statistical analysis
Statistical analysis GraphPadPrismprogrammever.4.0 carries out one factor analysis of variance inspection, and result is shown as mean+SD,
p﹤ 0.05 is for there being significant difference.
two, result
(1) Tianmu lake ethanol extract significantly reduces high fat induction apoE
-/-the body weight regulating liver-QI weight of mice.
Before experiment starts, mice is divided into 6 groups at random, and each group body weight is without significant difference.Feed high lipid food is after 12 weeks, model group apoE
-/-mouse Weight, liver weight average obviously raise (
p﹤ 0.01).Compared with model group, Tianmu lake ethanol extract obviously can reduce Mouse Weight regulating liver-QI weight, and its effect is obviously better than simvastatin (table 1).
Table 1 Tianmu lake is to high fat feed apoE
-/-the impact (means ± SD, n=8) of Mouse Weight, liver weight and liver index
| Group | Liver heavy (g) | Body weight (g) | Liver index (%) |
| CD | 1.20±0.17 | 29.12±1.15 | 4.12±0.15 |
| HFD | 1.50±0.43* | 33.28±5.58* | 4.51±0.20 |
| SV | 1.27±0.13 # | 29.71±1.96 # | 4.27±0.16 |
| TMQH-L | 1.15±0.17 ## | 29.12±2.12 # | 3.95±0.18 |
| TMQH-M | 1.20±0.23 # | 28.58±1.06 # | 4.20±0.21 |
| TMQH-H | 1.10±0.19 ## | 27.35±2.12 ## | 4.02±0.23 |
* p<0.05vsCD group;
# p<0.05,
## p<0.01vsHFD group.CD, blank group; HFD, hyperlipidemia model group; SV, simvastatin group; TMQH-L, Tianmu lake low dose group; TMQH-M, dosage group in Tianmu lake; TMQH-H, Tianmu lake high dose group.
(2) Tianmu lake ethanol extract is to apoE
-/-the impact of lipid of mice
As shown in Figure 1, high fat feed is after 12 weeks, model group apoE
-/-in mice plasma TC, LDL-C level all obviously raise (
p﹤ 0.01).After giving basic, normal, high dosage Tianmu lake ethanol extract, TC reduces 14.89%, 15.51% and 22.76% respectively.LDL-C level reduces 14.48%, 15.25% and 22.42% respectively.Meanwhile, the basic, normal, high dosage group HDL-C of Tianmu lake increases 8.02%, 16.25% and 51.86% respectively, and Tianmu lake on TG level without impact.
(3) Tianmu lake ethanol extract reduces oxidative stress level and inflammatory Cytokines Expression in blood plasma.
MDA is lipid peroxidation product, and it is the most frequently used Testing index of antimer internal oxidition stress level, and SOD is antioxidant.As shown in Table 2, feed high lipid food is after 12 weeks, and in blood plasma, MDA content obviously raises, and SOD is active obviously to be reduced.Compare with model group, in the basic, normal, high dosage group of Tianmu lake, MDA level reduces by 20.03%, 27.09% and 32.72% respectively; SOD activity raises 7.78% respectively, 9.67% and 12.75%(table 2).
Table 2 Tianmu lake is to apoE
-/-the impact (means ± SD, n=8) of SOD, MDA activity in mice plasma
| Group | SOD (U/ ml) | MDA(nmol/l) |
| CD | 183.92±19.05 | 25.92±7.13 |
| HFD | 164.65±20.61* | 36.95±4.81* |
| SV | 185.80±7.43 # | 22.26±1.50 ## |
| TMQH-L | 177.45±11.62 # | 29.55±4.18 ## |
| TMQH-M | 180.57±5.61 # | 26.94±3.89 ## |
| TMQH-H | 184.65±28.92 # | 24.86±3.65 ## |
* p<0.05
vscD group;
# p<0.05, ## p<0.01vshFD group.CD, blank group; HFD, hyperlipidemia model group; SV, simvastatin group; TMQH-L, Tianmu lake low dose group; TMQH-M, dosage group in Tianmu lake; TMQH-H, Tianmu lake high dose group.
In addition, inflammation is through in the atherosclerotic generation of blood vessel wall and evolution.Proinflammatory factor-tumor necrosis factor-alpha (TNF-α), can activate NF-κ B and the generation of stimulating activity oxygen in blood plasma.Inflammatory mediator important in blood plasma--interleukin-6 (IL-6), plays a significant role in vascular system.Acute stress albumen-C type reactive protein (CRP), can stimulate the expression of TNF-α and IL-6 under blood plasma inflammatory conditions.Elissa method is utilized to have detected the content of 3 kinds of important inflammatory factors in blood plasma in research.Compared to blank group, high fat diet induction enhances the expression of above-mentioned inflammatory factor, significant difference (
p<0.01).As seen from Figure 2, in model group blood plasma IL-6, TNF-α and CRP level significantly raise (
p<0.01), medication is after 12 weeks, and the basic, normal, high dosage group IL-6 of Tianmu lake declines 10.40%, 18.63% and 20.91% respectively; TNF-α declines 11.20%, 11.25% and 17.92%; CRP decline 12.41%, 23.45% and 18.62% respectively (
p<0.01 or
p<0.05).
(4) Tianmu lake significantly reduces apoE
-/-the formation of mouse aorta atherosclerotic plaque
As seen from Figure 3, blank group aorta structure is normal, almost without lipidosis and Mottling formation.High fat diet is after 12 weeks, and model group aorta inwall Lipid Plaque is formed significantly.Medication Tianmu lake is after 12 weeks, and basic, normal, high dosage group dose-dependently inhibits the deposition of lipid and the formation of speckle, wherein high dose group changed dramatically in the formation of artery plaque and aorta wall damage (
p<0.01) (Fig. 3 A, B, C).
Certainly, above-mentioned explanation is not limitation of the present invention, and the present invention is also not limited in above-mentioned citing, and the change that those skilled in the art make in essential scope of the present invention, remodeling, interpolation or replacement also should belong to protection scope of the present invention.
Claims (3)
1. the application of Tianmu lake ethanol extract in preparation treatment atheromatosis medicine.
2. the application of Tianmu lake ethanol extract according to claim 1 in preparation treatment atheromatosis medicine, it is characterized in that, the extracting method of described ethanol extract is: dried by Tianmu lake fruit room temperature, then 40 order coarse powder are ground to form, get 500g powder in the ethanol of 95% and extract, the mass volume ratio of described powder and ethanol is 1:10, ethanol extract is condensed into extractum after filtering, every gram of extractum is equivalent to 3.49g crude drug, then by extractum with 5% sodium carboxymethyl cellulose be mixed with 2.5g crude drug/ml respectively, 5.0g crude drug/ml, 10.0g crude drug/ml, 4 DEG C save backup.
3. the application of Tianmu lake ethanol extract in the exogenous atherosclerosis medicine of preparation treatment.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106674239A (en) * | 2016-12-21 | 2017-05-17 | 泰山医学院 | Viburnum sargentii branch and leaf lignan, extraction method and application |
| KR101994557B1 (en) * | 2017-12-28 | 2019-06-28 | 한국생명공학연구원 | Composition for preventing or treating metabolic syndrome comprising extracts of Viburnum stellato-tomentosum (Oerst.) Hemsl., a fraction thereof |
| WO2020251085A1 (en) * | 2019-06-13 | 2020-12-17 | 한국생명공학연구원 | Composition for prevention or treatment of metabolic syndrome comprising viburnum stellato-tomentosum extract or fraction thereof |
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| JP2010235468A (en) * | 2009-03-30 | 2010-10-21 | Uha Mikakuto Co Ltd | Banaba extract effective for activating lox-1 antagonist action and having ensured taste quality |
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Patent Citations (2)
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| JP2010235468A (en) * | 2009-03-30 | 2010-10-21 | Uha Mikakuto Co Ltd | Banaba extract effective for activating lox-1 antagonist action and having ensured taste quality |
| CN104286367A (en) * | 2014-10-19 | 2015-01-21 | 陆建益 | Health care ice-cream assisting to reduce blood fat and preparation method for ice-cream |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106674239A (en) * | 2016-12-21 | 2017-05-17 | 泰山医学院 | Viburnum sargentii branch and leaf lignan, extraction method and application |
| KR101994557B1 (en) * | 2017-12-28 | 2019-06-28 | 한국생명공학연구원 | Composition for preventing or treating metabolic syndrome comprising extracts of Viburnum stellato-tomentosum (Oerst.) Hemsl., a fraction thereof |
| WO2020251085A1 (en) * | 2019-06-13 | 2020-12-17 | 한국생명공학연구원 | Composition for prevention or treatment of metabolic syndrome comprising viburnum stellato-tomentosum extract or fraction thereof |
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