同型半胱氨酸影响Caco-2细胞线粒体功能的实验研究

Homocysteine induces mitochondrial dysfunction in Caco-2 cells

目的观察同型半胱氨酸(Hcy)对Caco-2细胞线粒体功能的影响。方法在Caco-2细胞中加入不同浓度(0、10、25、50、100、500、1 000μmol/L)Hcy作用不同时间(3、6、12、24 h),采用MTT法及检测乳酸脱氢酶活性,观察Hcy对Caco-2细胞生长活力的影响,检测丙二醛(MDA)、超氧化物歧化酶(SOD)、琥珀酸脱氢酶(SDH)、谷胱甘肽过氧化物酶(GSH-Px)、三磷酸腺苷酶(ATPase)水平及线粒体肿胀度观察Hcy对Caco-2细胞线粒体功能的影响。结果随着Hcy作用时间延长及作用浓度的增加,Caco-2细胞的生长活力受到明显抑制,MDA、SOD、GSH-Px水平较对照组明显增加,SDH、ATPase水平下降,线粒体肿胀度下降幅度较正常对照组降低。结论 Hcy具有损伤Caco-2细胞线粒体功能的作用,可能与Hcy氧化损伤有关。

Objective To investigate the effect of homocysteine （Hcy） on the growth of Caco-2 cells and the chan- ges of mitochondrial function. Methods Caco-2 cells were cultured and treated with Hcy in different concentra- tions （0,10,25,50,100,500,1 000 μmol/L） and different times （3,6,12,24 h）. MTT method and LDH activity were examined to understand the change of cell growth viability. MDA, SOD, SDH, GSH-Px, ATPase and mito- chondrial swelling degree were detected to understand the changes of mitochondrial function. Results After treat- ment with Hcy, the viability of Caco-2 cells were significantly inhibited with a dose and time dependent manner. Compared with the control group, the levels of LDH, MDA, SOD, GSH-Px increased significantly, and the levels of SDH and ATPase decreased. Compared to the normal control group, experimental groups mitochondria swelling de- gree was not decreased significantly. Conclusion Hcy has a significant inhibitory effect on the mitochondrial func- tion of Caco-2 cells in a dose-and time-dependent manner.