两种仙灵骨葆提取液对肝脏线粒体的作用

Comparison between Water and Ethanol Extracts of Xianlinggubao for Effects on Hepatic Mitochondria

目的通过考察仙灵骨葆对肝脏细胞线粒体功能来评价其是否有肝脏损伤作用。方法通过检测仙灵骨葆醇提液和水提液对大鼠原代肝脏细胞ATP含量,线粒体膜电位和线粒体特异荧光染料在线粒体内富集程度等线粒体功能和形态指标的影响,比较两种不同提取方法对肝脏线粒体的作用。结果仙灵骨葆醇提液在10~104倍稀释的情况下肝细胞线粒体状态均有显著变化,而水提液则仅在稀释10倍和100倍时线粒体状态才有明显改变。两者只有在稀释10倍（大约是临床剂量的50倍）时显著降低肝细胞ATP含量以及线粒体膜电位。结论仙灵骨葆两种提取方式中水提法与临床使用方式更为接近,这提示目前仙灵骨葆临床使用剂量有较宽的安全范围,而醇提法对肝细胞活力以及线粒体的形态和功能的影响明显强于水提法。

Objective To investigate whether XLGB can result in liver injury through the change of mitochondrial function index.Methods Primary hepatocytes isolated from SD rat were treated with serial dilution（1/10~1/10000） of XLGB ethanol and water extracts for 24 h. Cellular ATP content, mitochondria membrane potential（MMP） and accumulation of fl uorescent probe（TMRE） in mitochondria were used to compare and evaluate the potential liver injury induced by two extracts of XLGB.Results Only 1/10 dilution of water and ethanol extracts obviously decreased cellular ATP content and MMP in rat primary hepatocytes. Serial dilution of Ethanol extracts（1/10~1/10000） dose-dependently reduced the accumulation of TMRE in the mitochondria, while only 1/10 and 1/100 dilution of water extracts induced moderate reduction of TMRE accumulation. Conclusion In this study, ethanol extracts induced more obvious mitochondrial injury than water extracts, the latter is closer to the route of XLGB administration in human. In the serial tests of water extracts, the change of mitochondrial function was only induced by 1/10 dilution（about 50-fold clinical exposure in human）, indicating that XLGB may have a wide therapeutic window in clinical use.