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目的:研究Par-4反义寡核苷酸拮抗谷氨酸对PC12细胞中信号转导子和激活子3(STAT3)活性的下调作用及其抗凋亡意义。方法:脂质体将Par-4反义寡核苷酸转染PC12细胞。谷氨酸诱导PC12细胞凋亡。吖啶橙/溴化乙锭荧光染色观察PC12细胞形态。流式细胞分析评价凋亡百分率。Western blot测定Par-4的蛋白表达量,凝胶迁移改变实验测定STAT3的DNA结合力。结果:谷氨酸诱导PC12细胞中蛋白表达上调,Par-4反义寡核苷酸呈剂量依赖性地拮抗其上调(P<0.01)。谷氨酸诱导PC12细胞中STAT3的DNA结合力下调,Par-4反义寡核苷酸拮抗其下调(P<0.01)。Par-4反义寡核苷酸拮抗谷氨酸诱导的PC12细胞凋亡。如予Jak/STAT3信号通路阻断剂AG-490预处理,则其拮抗作用被下调。结论:Par-4反义寡核苷酸拮抗谷氨酸诱导的PC12细胞凋亡,其机制可能与STAT3活化有关。
AIM: To investigate the down-regulation of signal transducers and activator of transcription 3 (STAT3) activity and their anti-apoptotic effects by Par-4 antisense oligonucleotide against glutamate in PC12 cells. Methods: Liposomes were transfected into PC12 cells with Par-4 antisense oligonucleotide. Glutamate Induces PC12 Cell Apoptosis. Acridine orange / ethidium bromide staining was used to observe the morphology of PC12 cells. Flow cytometry was used to assess the percentage of apoptosis. The protein expression of Par-4 was determined by Western blot, and the DNA binding ability of STAT3 was determined by gel shift assay. Results: Glutamate up-regulated the protein expression in PC12 cells. Antisense oligonucleotides of Par-4 were up-regulated in a dose-dependent manner (P <0.01). Glutamate induced a down-regulation of STAT3 DNA binding in PC12 cells, and downregulation of Par-4 antisense oligonucleotides (P <0.01). Par-4 antisense oligonucleotide antagonizes glutamate-induced PC12 cell apoptosis. When pretreatment with Jak / STAT3 signaling pathway inhibitor AG-490, its antagonism was down-regulated. CONCLUSION: Par-4 antisense oligonucleotide antagonizes glutamate-induced PC12 cell apoptosis and its mechanism may be related to STAT3 activation.