1.Research on Composite Enzyme Assisted Hyperoside Extraction ofAcanthopanax senticosusand Its Process
Qiulian CHEN ; Manling ZHENG ; Enbo CAI ; Yan ZHAO ; Tianqi ZHAO ; Shumei TU ; Limin YANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(9):1866-1871
This study was aimed to optimize the extraction process of hyperoside from leaves ofAcanthopanax senticosusby compounding-enzyme method and orthogonal experiment. The hyperoside compound was regarded as standard and determined by HPLC. Based on the experiments of 4 factors including the enzyme amount, temperature, extraction time and PH values, the extraction process of hyperoside was determined by the orthogonal experiments and variance analysis. The results of single-factor experiments showed that different enzymes showed different effects on the enhance yield of hyperoside. The effects of different factors showed that the order of PH, neutral protease, temperature, time, pectinase, xylanase and cellulose was from strong to weak. Through orthogonal analysis, the optimum conditions were 2% pectinase, 2% xylanase, 0.5% neutral protease, and 0.5% cellulose, under the temperature of 30°C, extraction time of 10 min, and pH = 4.5. Under these conditions, the extraction rate was 1.84%. The yield was increased 107% compared with traditional process. It was concluded that the use of compounding enzyme can increase the yield of hyperoside, which possessed a lot of economic benefits.
2.Optimization of Extraction Process for Arctiin and Arctigenin inArctium lappaL. Based on Central Composite Design and Response Surface Methodology
Enbo CAI ; Wei WANG ; Caixia JIA ; Yan ZHAO ; Dalong WANG ; Xiangxiang LIU ; Dingrong YE ; Limin YANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(9):1828-1832
This study was aimed to optimize the extraction process of double-marker components for Arctium lappa L. The central composite design and response surface methodology was used. According to 3 main factors, the extraction rates of arctiin and arctigenin was used as evaluation indexes. Multiple linear regression and two-order polynomial equation were used. The binomial fitting model was performed in the optimization of arctiin and arctigenin extraction technology. The results showed that the indentified optimized extraction technology of arctiin and arctigenin was 70% ethanol, 24-fold, ultrasonic solvent extraction for 15 minutes. It was concluded that this technology was able to extract large amount of arctiin and arctigenin, which provided experiment evidences for arctiin and arctigenin preparation. It also provided references for the development and utilization of arctiin and arctigenin.
3.Study on Composite Enzyme Assisted Extraction of Hyperoside from Acanthopanax sessiflorus
Enbo CAI ; Jing XU ; Xiaoman ZHENG ; Yanan WANG ; Dalong WANG ; Manling ZHENG ; Xiangxiang LIU ; Yan ZHAO
Herald of Medicine 2016;35(6):636-639
Objective To extract hyperoside from the leaves of Acanthopanax sessiliflorus by complex enzyme method, and optimize the extraction process by orthogonal experiment. Methods Hyperoside was determined by HPLC. Effects of temperature,α-amylase, neutral protease and cellulase on extraction rate were detected by the orthogonal tests, and the optimum extraction condition of hyperoside from the leaves of Acanthopanax sessiliflorus was determined by complex enzyme method. Results The main influence factor was temperature,follows byα-amylase, neutral protease and cellulase according to orthogonal analysis.The best condition was as follows: dose of cellulase, neutral protease and α-amylase was 2%, 0. 5% and 3%, respectively, extract at temperature of 30 ℃for 10 min. Under this condition, the extraction rate of hyperoside in the leaves of Acanthopanax senticosus was 0.52%. Conclusion As compared with the traditional technics, compound enzyme increases the productivity of hyperoside.
4.Analysis to chemical constituents of Caulis of Schisandra and chemical pattern recognition
Chongxi ZHANG ; Jinggang LI ; Haibo XU ; Yanfang WANG ; Enbo CAI ; Dengli CONG
Chinese Traditional Patent Medicine 1992;0(08):-
AIM:To anylsis the chemical constituents of the Caulis of Schisandra from different harvesting time in Jiaohe prefecture for comprehensive utilization of shizandra berry's resource.METHODS:Dexyschisandrin,monosaccharide and oligosaccharide,polysaccharide were taken as statistical indicator and in combination with cluster analysis to compare among them.RESULTS:With colligational comparison,contents of climbing stem collected in November and July approached that of schisandra fruit.Deoxyschisandrin content was the highest in stem in July,polysaccharide content in November,oligosaccharide content in September.CONCLUSION:Chemical pattern recognition can play a role in comparing content of multicomponent of the Caulis of Schisandra and the best harvesting time.
5.A Research on the Anti-tumor Effects of Acid-Soluble Polysaccharides from Gloeostereum incamatum in H22 Tumor Bearing Mice
Hong WANG ; Yan ZHAO ; Enbo CAI ; Tolgor BAU ; Wei LI ; Yu LI ; Haiying BAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(1):142-148
This study aimed at exploring the inhibitory effect behind its mechanism on acid-soluble polysaccharides from G.incamatum in transplanted H22 tumor mice.Different indices,including tumor inhibitory rate,organ index of liver,thymus and spleen,IL-2,IFN-γ and TNF-α were detected for the evaluation of anti-tumor effects and the mechanism.Furthermore,HE staining and TUNEL assay were adopted to investigate the pathological changes of tumor tissue and cell apoptosis,respectively.As a result,the three dose groups of acidsoluble polysaccharides of G.incamatum successfully inhibited the proliferation of tumor cells,while organ indexes of spleen and thymus were improved and serum IL-2,IFN-γ and TNF-α increased.H&E staining and TUNEL assay showed the polysaccharides induced cell apoptosis,playing a significant role in the inhibition of tumor growth.In conclusion,acid-soluble polysaccharides of G.incamatum possessed significant anti-tumor effects,behind which the mechanism could be related to the regulation of immune regulation,cell apoptosis,and the protection of liver function.
6.Ginsenoside Rh4 induces apoptosis of human hepatocellular carcinoma HepG2 cells
Zi WANG ; Xiaoyan Lü ; Junnan HU ; Yan ZHAO ; Enbo CAI ; Shuangli LIU ; Wei LI ; Lianxue ZHANG
Chinese Journal of Pathophysiology 2017;33(8):1399-1404
AIM: To investigate the apoptosis and molecular mechanism of human hepatocellular carcinoma HepG2 cells induced by ginsenoside Rh4.METHODS: Human hepatocellular carcinoma HepG2 cells were treated with ginsenoside Rh4 at doses of 10, 20 and 40 μmol/L, and the inhibitory effect of ginsenoside Rh4 on HepG2 cell viability was measured by MTT assay.The apoptotic rate of HepG2 cells was analyzed by flow cytometry.The morphological changes of the HepG2 cells were observed by Hoechst 33258 and TUNEL staining.The expression of apoptosis-related proteins Bax, Bcl-2, caspase-3 and caspase-9 was determined by Western blot.RESULTS: Ginsenoside Rh4 promoted apoptosis of HepG2 cells in a dose-dependent manner.TUNEL and Hoechst 33258 staining showed that the cells appeared obvious shrinking, swelling and rupture after treated with ginsenoside Rh4 for 24 h.The results of Western blot showed that with the increasing concentrations of ginsenoside Rh4, the expression of pro-apoptotic proteins Bax, cleaved caspase-3 and caspase-9 increased, while anti-apoptotic protein Bcl-2 decreased gradually.CONCLUSION: Ginsenoside Rh4 induces apoptosis of human hepatocellular carcinoma HepG2 cells, and the main mechanism may be related to down-regulation of Bcl-2 and up-regulation of Bax, cleaved caspase-3, and caspase-9.