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Ampelopsis grossedentata as a traditional medicinal plant， has many pharmacological effects. Pharmacological effects and possible mechanisms are summarized in this paper based on the research progress of A. grossedentata at home and abroad in recent years， involving digestive system （treating liver injury and improving intestinal flora imbalance）， endocrine system （treating hyperglycemia， hyperlipidemia and hyperuricemia）， cardiovascular system （treating hypertension， vascular injury， myocardial ischemia-reperfusion injury and myocardial hypertrophy）， immune system （anti-inflammatory and anti-tumor） and nervous system （adjuvant treatment for chronic degenerative diseases）. By summarizing the animal acute toxicity test， long-term toxicity test and cell test results of A. grossedentata in the literature， it is confirmed that its safety is good.
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Objective To study the content differences of Dihydromyricetin in Ampelopsis grossedentata from different origins, parts and processing techniques, and improve the therapeutic effects by optimizing the compatibility of Ampelopsis grossedentata with Fructus arctiine. Methods The HPLC separation of Dihydromyricetin was performed on an Agilent ZORBAX SB-C18 column with methanol, 0.05 % phosphoric acid (30∶70) as the mobile phase. The flow rate was 1 ml/min, the detection wavelength was 291 nm, and the column temperature was 25 °C. Compatibility efficacy verification was performed with the inflammation model caused by cotton ball implantation in rats and ear swelling in mice. The net granulomatosis in the rats with cotton ball implantation and the swelling rate of mouse ears were recorded. Results Dihydromyricetin had a good liner recovery between 0.019 9-0.318 mg/ml (r=0.999). The extracted recovery was in the range of 95.04 %-100.4 %. The sample was stable within 24 h. This method had good repeatability. The combination of optimized high-dose Ampelopsis grossedentata with Fructus arctiine resulted in significantly lower net granuloma in rats and ear swelling rate in mice compared to the blank control group. Conclusion This method is simple and accurate. The content of dihydromyricetin varies greatly with different origins, parts, and processing techniques. Among them, the natural sun-dried vine tea in Jiangkou County, Guizhou Province has the highest content. The combination use of Ampelopsis grossedentata and Fructus Arctiine can significantly alleviate the pharyngeal symptoms, reduce the degree of inflammation, and achieve the therapeutic effect of clearing pharynx.
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OBJECTIVE：To study the spectrum-effect relationship of HPLC finger print of different polar parts of Ampelopsis grossedentata with its in vivo anti-inflammatory effect. METHODS ：A. grossedentata was reflux extracted with 70％ ethanol，then extracted with petroleum ether ，chloroform，ethyl acetate and water saturated n-butanol；or it was directly decocted with water and then concentrated to obtain different polar parts. The xylene-induced mice ear swelling model was established ；using dexamethasone as positive control ，anti-inflammatory activity of different polar parts of A. grossedentata was investigated. Fingerprints of different polar parts of A. grossedentata were established by HPLC. The determination was performed on Poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1 mL/min. The column temperature was 25 ℃. The detection wavelength was set at 365 nm，and sample size was 5 μL. The grey ralational analysis method was used to analyze the spectrum-effect relationship of HPLC fingerprint common peaks of different polar parts of A. grossedentata with its anti-inflammatory effect. The correlation coefficient and correlation degree were calculated and ranked. RESULTS：Anti-inflammatory experiment showed that the anti-inflammatory effects of 70％ ethanol extraction part ，ethyl acetate extraction part and water extraction part were the most significant （inhibitory rates of ear swelling were 54.07％，30.54％， 30.45％）. Five common peaks were determined in HPLC fingerprints of different polar parts from A. grossedentata . The spectrum-effect analysis results showed that the correlation of5 common peaks were higher than 0.6；among them ，peak 3 and peak 2 （dihydromyricetin） had the strongest anti- inflammatory effect ，and their correlation degrees were both mail：123745789@qq.com greater than 0.8. CONCLUSIONS ： The anti-inflammatory effect of A. grossedentata on xylene-induced ear swelling in mice is the result of multi-comp onent synergy ； unknown substance of peak 3 and dihydromyricetin may be the main active components of A. grossedentata .
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Objective: To study inhibitory effect of total flavonoids from Ampelopsis grossedentata (TF) on transplanted tumors of human hepatocellular carcinoma in nude mice, and predict that its mechanism may be related to relevant factors regulating phosphatidylinositol 3 kinase (PI3K)/protein kinases B(Akt)/p53 pathway in apoptosis. Method: The nude mice transplanted BEL-7404 hepatoma model was established and divided into model group, 5-fluorouracil (5-FU) group (1.0 g·L-1) and TF (30, 15, 7.5 g·L-1) groups. Nude mice were put to death after two weeks of administration. The tumor tissues were excised, and tumor inhibition rate (IR) and relative tumor proliferation rate (T/C) were calculated. Reverse transcription PCR(RT-PCR) was used to detect PI3K, Akt1, p53 gene(p53), Caspase-3, B cell lymphoma/lewkmia-2 (Bcl-2), Bcl-2 associated X protein (Bax) mRNA expressions, immunohistochemical method was used to detect expressions of relevant proteins PI3K, Akt1, p53, Caspase-3, Bcl-2, Bax. Result: The establishment of xenograft tumor in mice showed that TF was administered orally once per day for two consecutive weeks. IRs were 53.26%, 35.94%, and 26.74%, respectively. T/Cs were 59.74%, 69.66%, and 84.82%, respectively. RT-PCR experiments showed that compared with model group, when TF concentration was 30 g ·L-1, mRNA expressions of PI3K, Akt1, and Bcl-2 were significantly down-regulated, and mRNA expressions of tumor suppressor genes p53, Capsase-3, and Bax were significantly up-regulated. Immunohistochemical method results showed that compared with model group, at TF concentrations of 30, 15 g·L-1, all PI3K, Akt1, Bcl-2 protein expressions were significantly down-regulated, while p53, Capsase-3, Bax protein expressions were significantly increased. Conclusion: TF has an obvious anti-liver cancer activity in vivo. Its mechanism may be correlated with up-regulation of expressions of p53, Caspase-3, and activation of apoptosis PI3K/Akt/p53 pathway, thereby inhibiting Bcl-2, increasing expression of Bax, and promoting hepatocellular apoptosis.
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AIM To determine effective components of Ampelopsis grossedentata (Hand.-Mazz.) W.T.Wang under different breeding modes and to analyze the antioxidant enzyme activities.METHODS Single factor and orthogonal tests were adopted in screening the optimal extraction process of antioxidant enzymes and analyzing antioxidant enzyme activities of A.grossedentata under common cutting,two-phase cutting and tissue culture.Solvent extraction method was adopted in extracting the effective components of three breeding modes in stem leaves of A.grossedentata,whose contents were determined.RESULTS In three breeding modes,the content of polysaccharides in tissue culture seedling was the highest,while the contents of flavonoids and polyphenols in the two stage cutting seedlings were the highest.The optimal extraction process of antioxidant enzymes was determined to be 6.0 for pH,20 ℃ for processing temperature and 20 min for processing time.Antioxidant enzyme activities were in sequence of two stage cutting seedlings ＞ common cutting seedling ＞ tissue culture seedling.CONCLUSION Considering the contents of effective components and antioxidant enzyme activities,two stage cutting method is the best among 3 breeding modes.
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OBJECTIVE: To investigate the effects of total flavones of Ampelopsis(TFAG) on blood glucose and its antioxidant capacity. METHODS: The TFAG were extracted by ultrasonic assisted extraction and purified by D-101 macroporous resin. The effect of TFAG on the activities of α-glucosidase and α-amylase were determined in vitro. Diabetes mellitus(DM)mouse models were established by induction of streptozotocin(STZ). And the mice were daily treated by different dose of TFAG (100, 200, 400 mg·kg-1) for 15 d. In order to determine the blood glucose, blood was collected every 3 d. After administration of drug for 16 d, the blood was collected from the eyes and serum insulin and other biochemical indicator were measured. Hematoxylin and eosin stain will be used for histological analysis after collection of pancreatic tissues. Meanwhile, the amount of SOD, CAT and MDA in liver were measured. RESULTS: TFAG could significantly inhibit the activity of α-glucosidase and α-amylase, and that TFAG had a better inhibitory activity on α-glucosidase than α-amylase. The results of in vivo assay showed that TFAG could significantly reduce the levels of blood glucose, LDL, TC and TG, and increase the concentration of insulin and HDL in diabetic mice. HE staining showed that TFAG had some repair effects on pancreatic islet in diabetic mice. It was found that TFAG significantly decreased the levels of MDA, but instead it significantly increased the activities of SOD and CAT in liver of diabetic mice. CONCLUSION: The TFAG has significant effect on the hypoglycemic and antioxidant activity, and promote repair of pancreatic islets. Collectively, TFAG are valuable raw material with medicinal and edible properties and high development value.
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Objective: To screen reference genes for real time quantitative PCR (qRT-PCR) research in Ampelopsis grossedentata. Methods: On the basis of the conserved sequences among plant species, six candidate reference genes (including Actin, 18 S-rRNA, GAPDH, α-Tubulin, β-Tubulin, and UBQ) were cloned from A. grossedentata by RT-PCR in this study. The expression stability of each reference gene in different tissues (shoot tip, young leaf, mature leaf, old leaf, stem, and root) were analyzed by three softwares (GeNorm, NormFinder, and BestKeeper), followed by validation of the expression pattern of AgPAL by qRT-PCR. Results: Actin, 18 S-rRNA, and GAPDH expressed most stably in all samples and were suitable for reference genes, which were further confirmed by the transcript level analysis result of AgPAL in different tissues. Conclusion: This is the first report on the screening and validation of reference genes for qRT-PCR in A. grossedentata, which benefits future studies on gene expression in this species.
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In recent years, researches on the antibacterial experiments of single herb, extracts, monomer ingredients and the compound preparations have confirmed that a variety of Chinese meteria medica have good antibacterial effects, and they are not easily produced drug resistance. Ampelopsis grossedentata, which can be used as medicine and food, contains various biological ingredients. Among them, flavonoids have desired antibacterial effect and synergistic effect when combined with antibiotics. In this review, antibacterial effects of A. grossedentata were summarized from domestic and foreign study literatures in the recent decade, including the main antibacterial components, the antimicrobial effects, and mechanism of this medicinal plant, aming to provide reference for the studies on the physiological and pharmacological functions, clinical application, and product development of A. grossedentata.
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OBJECTIVE:To optimize the purification technology of total flavones from Ampelopsis grossedentala with macro-porous resins. METHODS:4 kinds of macroporous resins for the purification of total flavones from A. grossedentala were screened by using drug-loading amount,desorption rate,recovery and purification rate as indicators. Single factor test and central composite design-response surface methodology were used to optimized eluant mass fraction,adsorption time,flow rate of eluant,eluant pH and other factors of purification technology,and validation test was also conducted. RESULTS:D-101 macroporous resin was the best. The optimal condition was as follows as the concentration of sample solution 2 mg(by extract weight)/ml,the volume of sam-ple solution 1.1 BV,ethanol 86.0%,adsorption time 36.7 min,flow rate of eluant 3.81 BV/h,pH 7. In validation test,mass frac-tion of total flavones increased from 66.83% to 85.00% in validation test(RSD=0.15%,n=3),and were close to predicted val-ue(85.08%). CONCLUSIONS:Central composite design-response surface methodology is feasible and stable for the optimization of purification technology of total flavones from A. grossedentala with macroporous resins.
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Objective: A new quality evaluation method using HPLC fingerprint coupled with quantitative analysis of multi-components by single marker method was developed for the quality control of Ampelopsis grossedentata. Methods: HPLC analysis was performed on an Agilent Eclipse Plus C18 (250 mm × 4.6 mm, 5 μm) column using methanol-0.05% aqueous solution of phosphoric acid as mobile phase at flow rate of 0.8 mL/min. The column temperature was set at 50 ℃ and the detection wavelength was at 258, 292, and 369 nm. The quality analyses of samples from different origins were accomplished by cluster analysis and principal component analysis using SPSS 19.0 statistical software. Results: The HPLC fingerprint of A. grossedentata from different regions was established. Twenty-four common peaks were found in the HPLC fingerprint of 14 samples, three important common components were identified as dihydromyricetin, myricitrin and myricetin. Further, the quantitative analysis of multi-components by single marker method of these contents was established. Conclusion: The method of HPLC fingerprint combined with quantitative analysis of multi-components by single marker method can be used for the quality control of A. grossedentata.
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OBJECTIVE: To study the chemical constituents of Ampelopsis grossedentata (Hand-Mazz) W.T. wang. METHODS: The chemical constituents were isolated and purified with silica column chromatography, gel chromatography, etc. Their structures were identified by physicochemical properties and various spectroscopic methods including NMR spectrum, MS, UV, etc. RESULTS: Sixteen compounds were isolated and elucidated as dihydromyricetin(1), physcion(2), quercetin(3), myricetin(4), taxifolin(5), kaempferol(6), dihydrokaempferol(7), myricetin-3-O-L-rhamnoside(8), afzelechin(9), bellidifod-in(10), quercetin-3-O-α-L-rhamnopyranoside(11), myricetin-3'-O-β-D-xylopyranoside(12), ampelopsin(13), myricetin-3-O-β-D-glucoside(14), astragalin(15) and myricetin-3-O-β-D-galactopyranoside(16). CONCLUSION: Compound 10 is reported from the plants in genus Ampelopsis for the first time. Compounds 5,6,7,9,12 and 15 are isolated from this plant material for the first time.
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Objective To observe the effect of total flavonoids of ampelopsis grossedentata on blood lipid and hemorheology in atherosclerosis rats .Methods The atherosclerosis model in rats was made by freeing high grease food and intraperitoneal injection of vitamin D3 ,total flavonoids of ampelopsis grossedentata was gave by gavage ,and the level of serum lipid and blood rheology were tested 24 weeks later .Results Total flavonoids of ampelopsis grossedentata could obviously decreased the level of TG ,TC ,LDL , and increased HDL(P<0 .05) .It could significantly decrease whole blood viscosity ,plasma viscosity and hematocrit (P<0 .05) . Conclusion Total flavonoids of ampelopsis grossedentata can adjust fatty substance metabolism ,improve hemorheology of athero-sclerosis rats ,and has therapeutic effect for atherosclerosis .
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Objective:To study the preventive and health effects of Ampelopsis grossedentata on hyperlipidemia. Method:Thirty Wistar rats were divided into three groups:control group, model group and test group. The control group was fed normal diet, while the model and test groups were fed high fat diet.The control and model group were given water while the test group was given water with 20% Ampelopsis grossedentata for 15 w. Serum lipids, hemorrheological indices, myocardial enzymes and IL-6 were measured. Results: At the end of experiment, serum TC, TG, LDL-C, and apoB/apoA, serum and myocardial MDA, myocardial enzymes activities, hemorrheological indices and serum IL-6 were obviously higher in model group while the activities of serum and myocardial SOD and GSH-Px were lower. In test group. serum TC, TG and apoB/apoA levels, serum and myocardial MDA, myocardial enzymes activities, hemorrheological indices, and serum IL-6 were significantly lower than those in model group, but HDL-C, SOD and GSH-Px activities higher. Conclusion:Ampelopsis grossedentata could significantly decrease blood lipids and hemorrheological indices of rats fed with high fat diet, and protect myocardial cells fromoxidation, and as a result to prevent hyperlipidemia and cardiovascular disease.
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OBJECTIVE:To investigate the inhibitory effect of the total flavoniods of Ampelopsis grossedentata on human hepatoma carcinoma cells(HCC) in vitro.METHODS:The inhibitory effect of the total flavoniods of Ampelopsis grossedentata extract on human hepatoma carcinoma BEL7404 cells in vitro was analyzed by MTT,cell growth curve method and colony formation method,respectively.RESULTS:The total flavonoids of Ampelopsis grossedentata significantly inhibited the growth of BEL 7404 cells in vitro.MTT assay showed that the IC50 was 28.59 ?g?mL-1;the cell growth curve measurement showed that the total flavonoids of Ampelopsis grossedentata significantly inhibited the growth of BEL 7404 cells;the colony formation assay showed that the IC50 was 100% at a drug concentration of above 22.22 ?g?mL-1 and the IC50 was 58.05% at a drug concentration of 14.81 ?g?mL-1.CONCLUSION:The total flavonoids of Ampelopsis grossedentata have a remarkable inhibitory effects on the growth of BEL 7404 in vitro.
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Object To establish a method for the assay of myricetin in Ampelopsis grossedentata (Hand. Mazz.) W. T. Wang. Methods The determination was carried out with RP HPLC on Nova pak C 18 stainless steel column (150 mm ? 3 9 mm) with methanol∶water (40∶60) as the mobile phase, and detected at a UV wave length of 254 nm. Results The coefficient of variation was 2.763% with average recoveries=97.4%. The minimal detectable limit was 15 ng. Contents of myricetin in different parts of A. grossedentata from Hunan Province varied from 1.57% to 2.17%. Conclusion The method is rapid, simple, accurate and good for the determination of myricetin in A. grossedentata.
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AIM: To establish the determination of dihydromyricetin in stem,leaf of Ampelopsis grossedentata in different seasons from different habitat. METHODS : Novapak C_ 18 column (150mm?4.6mm,5?m) was selected as separation column at 25 ?C . Methanol-water-phosphoric acid (27∶73∶0.1) was used as a mobile phase at a flow rate of 1.0mL?min -1 . The peak of dihydromyricetin was detected at UV 290nm. RESULTS : The linearity of this method was good. The average recovery was 99.47%,RSD was 1.68%. The content of dihydromyricetin in leaf of Ampelopsis grossedentata collected in May was the highest and was three ～ four times the size of that in stem. CONCLUSIONS :The method is convient with a good separating degree and is useful basis for the develoment and utilization of Ampelopsis grossedentata.