| 引用本文: | 杨华蕊,赵艳雯,杨永寿,肖培云.美洲大蠊抗肝纤维化活性提取物的酶解制备工艺研究[J].中国现代应用药学,2019,36(16):1985-1991. |
| YANG Huarui,ZHAO Yanwen,YANG Yongshou,XIAO Peiyun.Study on the Enzymatic Hydrolysis Preparation of Active Extractives of Periplaneta Americana Against Hepatic Fibrosis[J].Chin J Mod Appl Pharm(中国现代应用药学),2019,36(16):1985-1991. |
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| 摘要: |
| 目的 优选中性蛋白酶酶解美洲大蠊脱脂膏制备抗肝纤维化活性提取物的工艺。方法 以美洲大蠊脱脂膏的水解度、活性提取物的得率和体外活性为评价指标，在单因素考察（酶用量、酶解时间、pH值、酶解温度、底物浓度）的基础上，采用L9（34）正交试验优化中性蛋白酶酶解制备抗肝纤维化活性提取物的工艺。结果 最佳酶解工艺为酶用量0.3%，酶解时间2 h，pH值8.5，酶解温度45℃，底物密度1.07 g·mL-1。工艺验证试验结果表明，美洲大蠊脱脂膏的水解度为15.8%，活性提取物的得率从0.54%提高到0.72%，其24，48，72 h的IC50值分别为（118.37±3.12），（115.16±2.48），（105.00±6.35）μg·mL-1。结论 研究得到美洲大蠊抗肝纤维化活性提取物的酶解制备工艺，该工艺得率较高，简单可行，为美洲大蠊抗肝纤维化药物的研发奠定基础。 |
| 关键词: 正交试验 酶解 美洲大蠊 抗肝纤维化 中性蛋白酶 |
| DOI：10.13748/j.cnki.issn1007-7693.2019.16.001 |
| 分类号:R285.5 |
| 基金项目:国家自然科学基金项目（81360634，81560634）；云南省科技计划项目（2015FA025） |
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| Study on the Enzymatic Hydrolysis Preparation of Active Extractives of Periplaneta Americana Against Hepatic Fibrosis |
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YANG Huarui1, ZHAO Yanwen1, YANG Yongshou2, XIAO Peiyun1
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1.College of Pharmacy and Chemistry, Dali University, Dali 671000, China;2.Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R & D, Dali 671000, China
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| Abstract: |
| OBJECTIVE To optimize the enzymatic hydrolysis technology of anti-hepatic fibrosis active extractive from the degreasing ointment of Periplaneta americana by neutral protease. METHODS The hydrolysis degree, yield of active extractive and in vitro activity were used as evaluation indexes. On the basis of single factor investigation(enzyme dosage, enzymatic time, pH, enzymatic temperature, substrate concentration), the L9(34) orthogonal experiment was used to optimize the neutral protease hydrolysis of degreasing ointment of Periplaneta americana to prepare the active extractive of anti-hepatic fibrosis. RESULTS The optimum condition were as follows:enzyme dosage was 0.3%, enzymatic time was 2 h, pH was 8.5, enzymatic temperature was 45℃ and substrate density was 1.07. The results of process validation showed that the hydrolysis degree of Periplaneta americana degreasing ointment was 15.8%, and the yield of active extractive increased from 0.54% to 0.72%. The IC50 values for 24, 48 and 72 h were (118.37±3.12), (115.16±2.48), (105.00±6.35)μg·mL-1, respectively. CONCLUSION The enzymatic hydrolysis process of the active extractive of Periplaneta americana against hepatic fibrosis is optimized. The optimized process is simple and feasible, which laid a foundation for the follow-up study of the active fraction of Periplaneta americana. |
| Key words: orthogonal experiment enzymatic hydrolysis Periplaneta americana anti hepatic fibrosis neutral protease |
