中国药理学通报Issue(9)：1287-1292,6.DOI:10.3969/j.issn.1001-1978.2014.09.022
NO水平下降通过氧化应激引起人胎盘滋养细胞凋亡
The levels of NO decrease induced apoptosis in human placental trophoblast cells through oxidative stress
摘要
Abstract
Aim To investigate the possible mecha-nisms of the levels of NO decrease induced apoptosis in human placental trophoblast cells. Methods Human placental trophoblast cells ( HTR-8 ) were cultured in 5 ml DMEM-F12 culture medium with 37℃ 5% CO2 . Then, the old culture medium was discarded and re-placed with 10,100,500,1 000 μmol·L-1 L-NAME, and the group without L-NAME was set as the control group, cultured for 48h. The effects of L-NAME on the survival of cells were detected by methylthiazolyldiphe-nyl tetrazolium bromide ( MTT); the content of NO in cells was tested by nitrate reductive enzymatic;trans-mission electron microscopy, flow cytometry analysis and Annexin-V FITC dyeing were used to test the effects of L-NAME on apoptosis in HTR-8 cells;restore Fe3+ colorimetric assay was applied for detection of to-tal antioxidant capacity ( T-AOC ) , xanthine oxidase for detection of superoxide dismutase ( SOD) activity, and thiobarbituric acid colorimetry for determination of content of MDA. Results Compared with the control group, the survival rate of HTR-8 cells and the levels of NO in 100,500,1 000 μmol·L-1 L-NAME group were significantly reduced(P<0.05,P<0.01). Flow analysis and Annexin-V FITC staining showed that L-NAME could induce cell apoptosis in a dose-dependent manner. The number of cell apoptosis was negatively correlated with the content of NO ( r = -0.5210 ) in HTR-8 cells. Transmission electron microscopy results showed that compared with the control group, the ex-perimental group's cell nucleus shape was irregular, nuclear pyknosis in irregular shape, the chromatin ag-glutination or side the collection, mitochondrial swell-ing or enrichment, crest fracture or dissolved, even vanished, forming the vacuole, especially in 100 μmol ·L-1 L-NAME group, the apoptotic bodies obviously appeared. At the same time, T-AOC, SOD levels in HTR-8 cells decreased ( P <0.05 ) , and the MDA content increased ( P<0.05 ) . The number of cell ap-optosis was negatively correlated with the level of T-AOC ( r= -0.3212 ) , SOD ( r= -0.2779 ) in HTR-8 cells , while positively correlated with the content of MDA(r=0.2807). Conclusion Oxidative stress may play an important role in the levels of NO decrease in-duced apoptosis in human placental trophoblast cells.关键词
一氧化氮/N-硝基-L-精氨酸甲酯/人胎盘滋养细胞/凋亡/氧化应激/妊娠期高血压疾病Key words
nitric oxide/N- nitro-L-arginine methyl ester ( L-NAME)/human cytotrophoblast cells/apopto-sis/oxidative stress/hypertension disorder complica-ting pregnancy分类
医药卫生引用本文复制引用
王艳华,韩学波,杨晓玲,姜怡邓,张慧萍,田珏,周龙霞,陈久凯,马文斌,孔繁琪,赵丽,刘现梅..NO水平下降通过氧化应激引起人胎盘滋养细胞凋亡[J].中国药理学通报,2014,(9):1287-1292,6.基金项目
国家自然科学基金资助项目( No 81260105) （ No 81260105）
宁夏高等学校科学研究项目(No NGY2013054) （No NGY2013054）
