反式油酸对H9c2心肌细胞增殖抑制及诱导凋亡作用的研究

目的研究反式油酸(9t-C18:1)对H9c2心肌细胞增殖抑制及诱导凋亡作用并探寻其可能的机制。方法体外培养H9c2大鼠心肌细胞,9t-C18:1高、中、低剂量组及阴性对照组(NC)分别作用于随机分组的H9c2细胞。采用CCK-8法检测细胞增殖能力;AO-EB染色后观察细胞形态学变化;流式细胞仪检测细胞内活性氧(ROS)及细胞凋亡情况;实时定量PCR(QRTPCR)法检测Bcl-2、Bax基因表达,免疫荧光染色后流式细胞仪检测Bcl-2、Bax蛋白表达。结果荧光显微镜下可见,9t-C18:1作用后的H9c2细胞出现典型的凋亡形态学特征;与NC组比较,随着9t-C18:1剂量的增加细胞增殖抑制作用明显,ROS、细胞凋亡率显著升高,Bcl-2基因及蛋白表达下调,Bax基因及蛋白表达上调(P <0. 05,P <0. 01)。结论 9t-C18:1能抑制H9c2细胞增殖并诱导其凋亡,其机制可能与促进细胞线粒体凋亡通路有关。

Effect of Trans-oleic Acid on Proliferation Inhibition and Apoptosis Induction in H9c2 Cardiomyocyte

OBJECTIVE To investigate the effects and possible mechanism of trans-oleic acid( 9 t-C18: 1) on proliferation inhibition and induction apoptosis in H9c2 cardiomyocyte. METHODS H9c2 rat cardiomyocytes were cultured in vitro. High,medium and low( 600,300,150 μmol·L-1) dose of 9 t-C18: 1 groups and the negative control( NC) group were administered to H9c2 cardiomyocytes. The effect of 9 t-C18: 1 on cell proliferation was tested using cell counting kit-8( CCK-8) assay. Morphological changes of cells were observed by AO-EB staining. Intracellular reactive oxygen species( ROS) and apoptosis were detected by flow cytometry.The expression of Bcl-2 and Bax genes was detected by quantitative real time-polymerase chain reaction( QRT-PCR). The expression of Bcl-2 and Bax protein was determined by flow cytometry after immunofluorescence staining. RESULTS The typical morphological characteristics of apoptosis were observed by fluorescence microscope. The result of CCK-8 assay indicated that 9 t-C18: 1 have an certainly inhibitory effect on the proliferation of H9c2 cells. ROS level and apoptosis rate were significantly increased. Bcl-2 gene and protein expression were down-regulated,and Bax gene and protein expression were up-regulated,compared with NC group( P < 0. 05,P < 0. 01). CONCLUSION 9 t-C18: 1 can inhibit the proliferation and induce the apoptosis of H9c2 cardiomyocyte,and its mechanism may be related to promoting the mitochondrial apoptotic pathway.