鳖甲水提物W-A-80在模拟胃肠液中消化情况分析

Digestion Analysis of Water Extract W-A-80 from Trionycis Carapax in Simulated Gastrointestinal Fluid

目的:研究鳖甲水提物W-A-80在模拟胃、肠环境中消化情况,探讨其体内转化形式。方法:采用水提醇沉法制备鳖甲水提物W-A-80,模拟胃肠液环境,于37℃消化。以寡肽产率为指标,通过双缩脲反应-酶联免疫检测仪法考察胃蛋白酶和胰蛋白酶于不同酶底比、酶解时间的寡肽(截留相对分子质量<3 500)产率变化;平衡透析法测定模拟胃肠液消化后所得寡肽与体外小鼠肝组织的蛋白结合率。结果:确定酶解条件为人工胃液酶底比1∶10,于37℃酶解3 h,不同质量浓度寡肽与肝组织的蛋白结合率分别为31.63%,19.93%,9.65%;人工肠液酶底比1∶100,于37℃酶解11 h,不同质量浓度寡肽与肝组织的蛋白结合率分别为48.75%,47.43%,22.72%;均明显高于空白对照的7.00%,5.31%,4.60%。结论:鳖甲水提物W-A-80经模拟胃肠液消化有大量寡肽产生,且活性显著强于原液,推测寡肽可能是鳖甲抗肝纤维化的物质基础。

Objective： To explore in vivo transformation form of aqueous extract W-A-80 from Trionycis Carapax through investigating its digestion in simulated gastric fluid（SGF） and simulated intestinal fluid（SIF）.Method： Aqueous extract W-A-80 from Trionycis Carapax was prepared by water extraction and alcohol precipitation method,it was digested at 37 ℃ in SGF and SIF.With yield of oligopeotides as index,effect of different enzyme-substrate ratios and reaction time of pepsin and trypsin on yield change of oligopeptides（interception relative molecular mass 3 500） was investigated by enzyme-linked immunosorbent detector-biuret reaction;Equilibrium dialysis was adopted to determined protein binding rate of prepared oligopeptides with liver tissue of mice in vitro.Result： Optimum enzymatic hydrolysis conditions were determined that digested 3 h with pepsin enzyme substrate ratio of 1∶ 10 at 37 ℃ in SGF and 11 h with trypsin enzyme substrate ratio of 1∶ 100 at 37 ℃ in SIF;Protein binding rates of different concentrations of oligopeptides with liver tissue in mice were 31.63%,19.93%,9.65% and 48.75%,47.43%,22.72%,respectively,both significantly more than that of blank control group which were 7.00%,5.31%,4.60%.Conclusion： A large amount of oligopeptides generated when aqueous extract W-A-80 was digested in simulated gastrointestinal fluid,and their activity were significantly stronger than undisposed,so we speculated oligopeptides may be materials basis of Trionycis Carapax for anti-liver fibrosis.