Evaluation of BCL-2 and BAX Genes Expression in Hyperglycemia-Induced NIH Cells

Sher Zaman
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Abstract

The incidence of Diabetes Mellitus (DM) type I and type II is very high all over the world. Excessive glucose levels and failure of one’s body to produce or manage glucose, trigger diabetes. Glucose is known to be responsive to NIH3T3 cells as it alters the expression of a range of genes associated with inflammation and apoptosis. In this study, the toxic effect of glucose was evaluated on NIH3T3 fibroblasts. Cells (NIH3T3) were cultured in media (DMEM), supplemented with 10% FBS and 1% Penicillin-Streptomycin. MTT assay was performed to check the toxic effect of glucose. NIH3T3 cells were treated with high glucose (30mM) for 24 hours. Trizol was used to extract the RNA followed by PCR reactions for gene expression analysis. Glucose treatment for 24 hours, modulated the expression of BCL-2 and BAX genes. The expression of BCL-2 was reduced while a significant increase was noticed in the expression BAX gene. Our results illustrated that glucose has some toxic effects on NIH3T3 cells. Glucose induces apoptosis by upregulating BAX and down-regulating BCL-2 expressions.
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BCL-2和BAX基因在高血糖诱导的NIH细胞中的表达
1型和2型糖尿病的发病率在世界范围内都很高。葡萄糖水平过高,身体不能产生或控制葡萄糖,会引发糖尿病。已知葡萄糖对NIH3T3细胞有反应,因为它改变了一系列与炎症和凋亡相关的基因的表达。本研究评估了葡萄糖对NIH3T3成纤维细胞的毒性作用。细胞(NIH3T3)在培养基(DMEM)中培养,培养基中添加10%胎牛血清和1%青霉素-链霉素。采用MTT法检测葡萄糖的毒性作用。NIH3T3细胞用高糖(30mM)处理24小时。用Trizol提取RNA, PCR反应进行基因表达分析。葡萄糖处理24小时后,BCL-2和BAX基因的表达发生变化。BCL-2表达降低,BAX基因表达显著升高。我们的研究结果表明,葡萄糖对NIH3T3细胞有一定的毒性作用。葡萄糖通过上调BAX和下调BCL-2表达诱导细胞凋亡。
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