百草枯致人肾小管上皮细胞凋亡及抗凋亡的PCR基因芯片研究  

The Investigation of the PCR Gene Chip of Apoptosis and Anti-apoptosis Caused by Paraquat to Human Kidney Tube Epithelial Apoptosis

作　　者：李倩 罗毅沣[1] 张剑锋[1] Li Qian;LUO Yi-feng;ZHANG Jian-feng(Department of Emergency,The Second Affiliated Hospital of Guangxi Medical University,Nanning 530007,P.R.China)

机构地区：[1]广西医科大学第二附属医院急诊科

出　　处：《岭南急诊医学杂志》2019年第5期411-414,430,共5页Lingnan Journal of Emergency Medicine

基　　金：广西自然科学基金项目（2017GXNSFAA198249）

摘　　要：目的:从转录组学角度对人肾小管上皮细胞的凋亡及抗凋亡基因表达变化机制作初步探讨。方法:人肾小管上皮细胞体外培养,用CCK8试剂盒检测百草枯中毒对人肾小管上皮细胞的毒性作用,测定半数有效剂量的浓度。实验组使用一定浓度百草枯处理人肾小管上皮细胞,对照组使用无血清培养基,均培养24小时。使用光镜观察,并用流式细胞仪分析确定凋亡存在,收集两组细胞提取总RNA。采用人凋亡信号通路PCR芯片检测两组细胞的相关基因表达情况,并使用qPCR验证差异基因CASP5、CD154、Bcl-2A1的表达情况。结果:百草枯可致人肾小管上皮细胞的CASP5凋亡基因及CD154和Bcl-2A1抗凋亡基因表达升高(P<0.05);用qPCR验证的结果与上述结果趋势一致。结论:CASP5可能在百草枯致人肾小管上皮细胞凋亡的过程中发挥其促凋亡的作用,而CD154及Bcl-2A1可能在百草枯致人肾小管上皮细胞抗凋亡相关的机制发挥作用。Objective:To make a preliminary discussion on the apoptosis of human renal tubular epithelial cells and the expression of anti-apoptotic gene expression from the perspective of transcriptomics.Methods:CCK8 kit was used to detect the toxicity of paraquat poisoning on human renal tubular epithelial cells,and the concentration of half effective dose were determined.The experimental group was treated by human renal tubular epithelial cells with a certain concentration of paraquat,and the control group was used serum-free medium for 24 hours.Light microscopy was used and flow cytometry analysis was used to determine the presence of apoptosis,and two groups of cells were collected for total RNA extraction.The human apoptotic signaling pathway PCR chip was used to detect the expression of related genes in the two groups of cells,and qPCR was used to verify the differential genes.Results:Paraquat could induce the expression of CASP5 apoptosis gene and CD154 and Bcl-2 A1 anti-apoptotic gene in human renal tubular epithelial cells(P<0.05).The results verified by qPCR are consistent with the above results.Conclusion:CASP5 may play a pro-apoptotic role in the process of apoptosis of human renal tubular epithelial cells induced by paraquat,and CD154 and Bcl-2 A1 may play a role in anti-apoptosis-related mechanisms of human renal tubular epithelial cells induced by paraquat.

关 键 词：百草枯 急性肾损伤 人肾小管上皮细胞 凋亡 抗凋亡 

分 类 号：R73[医药卫生—肿瘤]