泽泻不同提取物对SGC-7901细胞线粒体能量代谢的影响

Effect of Rhizoma Alismatis Extracts on SGC-7901 Cell Mitochondrial Energy Metabolism

目的比较泽泻不同提取物对细胞线粒体能量代谢的影响。方法分别制备泽泻水提取物(WRE)、乙醇提取物(ERE)、超临界CO2萃取物(SFE),以SGC-7901胃癌细胞为研究对象,阳离子荧光羰花青染料JC-1染色,采用倒置荧光显微镜观察结合流式细胞仪检测3种提取物对细胞线粒体膜电位的影响;应用酶标仪检测3种提取物对琥珀酸脱氢酶(SDH)活性的影响。结果红色荧光减弱(或绿色荧光增强)的程度:ERE<SFE<WRE。流式细胞仪检测结果显示,正常对照组、WRE组、ERE组、SFE组的单阳细胞比例分别为3.5%、14.8%、9.8%和17.3%,与正常对照组比较,3组提取物单阳细胞比例均有增加,差异有统计学意义(P<0.01),与ERE组比较,WRE组单阳细胞比例更高(P<0.01)、SFE组单阳细胞比例最高(P<0.01);SDH检测结果显示,SFE组对细胞SDH活性有抑制作用,而ERE组和WRE组均有不同程度的促进作用,且ERE组对SDH的促进作用大于WRE组;与正常对照组比较,ERE组和WRE组的SDH活性差异有统计学意义(P<0.01,P<0.05);与ERE组比较,WRE组、SFE组的SDH活性差异有统计学意义(P<0.05)。结论泽泻醇提物对线粒体膜电位的损害较小,对SDH活性促进作用较显著,提示泽泻醇提取物具有较明显改善线粒体能量代谢的作用。

Objective To observe the effect of Rhizoma Alismatis （RA） extract on cell mitochondrial energy metabolism. Methods RA extracts were prepared by the water reflux extraction （WRE）, ethanol reflux extraction （ERE） and the supercritical CO2 fluid extraction （SFE）, respectively. Human gastric carcinoma SGC-7901 cells were treated with the above three extracts, and then dyed by cation fluorescence carbocyanine JC-1. The mitochondrial membrane potential of SGC-7901 cells was determined by the inverted fluorescence microscope and flow cytometer, and the succinate dehydrogenase （SDH） activity was detected by microplate reader. Results The red fluorescence weakening （or green fluorescence strengthen） in the ERE, SFE, WRE groups was in increasing sequence, ERE〈SFE 〈 WRE. Single positive cell propotions of normal control（NC）, WRE, ERE, SFE groups were 3.5 %, 14.8%, 9.8 %, 17.3 %, respectively. Compared with the NC group, single positive cell proportions of WRE, ERE, SFE groups were inereased（P 〈 0.01）, and SFE group had the highest proportion（P 〈 0.01）. The result of SDH detection showed that SFE inhibited the activity of SDH, but the ERE and WRE promoted the activity of SDH （P 〈 0.01, P 〈 0.05 compared with normal control group） , and the effect of ERE was stronger than that of WRE （P 〈 0.05）. Conchlsion Ethanol extract of Rhizoma Alismatis has less damage on mitoehondrial membrane potential of SGC- 7901 cells, but can promote SDH activity, indicating that ethanol extract could improve the mitochondrial energy metabolism, and this will lay the foundation for investigating mitoehondrial energy metabolism in vivo.