不同方法水蛭提取液对体外培养LEC影响的实验研究

Effect of different extracting solutions of Hirudo on bovine lens epithelial cells cultivated in vitro

目的:观察不同方法水蛭提取液对体外培养的晶状体上皮细胞(LEC)生长的影响,比较不同提取方法的作用强度。方法:以五种常用中药提取法(水醇双提法、乙醇回流法、水提法、水提醇沉法、渗滤法)提取水蛭;植块培养法体外培养牛LEC,各方法水蛭提取液组以等比组距设置药物浓度,分别作用于体外培养LEC各24、72h;以MTT法测定OD值,并计算细胞抑制率,以克贝尔法计算各提取液组24、72hID50值。结果:水蛭渗滤液24hID50与水蛭水醇双提液、水蛭乙醇回流液24hID50比较有显著性差异(P<0.05);水蛭水提醇沉液24hID50与水蛭乙醇回流液24hID50比较有显著性差异(P<0.05)。结论:五种水蛭不同方法提取液均能不同程度抑制体外培养LEC的生长,其中水提醇沉液、渗滤液作用更为明显。

Objective： To observe the effect of different extracting solutions of Hirudo on the proliferation of bovine lens epithelial cclls （LEC） cultivated in vitro, for comparing the strength deflect of different extracting solutions.Methods： Five kinds of conventional extracting solution of Hirudo （alcohol and water extracting, alcohol re.fluxing, water extracting, water extracting-alcohol precipitating, percolation filtration） were used in this study.Bovine lens anterior capsule was cultured in vitro with expiant culture method.The passage LEC was incubated for 48h, then exposed the ceils to different extracting solution in different concentrations for 24h and 72h. MTT method was used to detect the OD values and ID50 was calculated. Results： There was significant difference of 24hID50 as the percolation filtration group compared with the alcohol and water extracting group and the alcohol refluxing group （ P 〈 0.05）, and there was significant difference of 24hID50 as the alcohol and water extracting solution group compared with the alcohol re.fluxlng group also （ P 〈 0.05 ） . Conclusions： Five kinds of conventional extracting solution of Hirudo all can inhibit the proliferation of LEC cultivated in vitro, and inhibition of water extracting-alcohol precipitating solution and percolation filtration solution was more strong among them.