氯沙坦介导AngⅡ/AT1R信号通路减轻百草枯诱导肾小管上皮细胞损伤

Losartan Mediates AngⅡ/AT1R Signaling Pathway Reducing Human Kidney-2 Cell Damage in Acute Paraquat Poisoning Model

目的:探讨氯沙坦在减轻百草枯诱导人肾小管上皮细胞损伤的机制和效果。方法:体外培养人肾小管细胞并将其分为对照组、实验组和干预组,以CCK8方法测得百草枯和氯沙坦的半数有效剂量。干预组使用百草枯+氯沙坦干预24小时,制成细胞爬片使用HE、TUNEL染色镜下查看细胞情况;采用RT?PCR技术测定AngⅡ、AT1R、IL?1C、IL?6和TNF?α的变化;采用WestenBlot技术检测AT1R。结果:实验组细胞可见空泡变性与凋亡增加,AngⅡ,AT1R,IL?1胞,IL?6和TNF?胞表达量增加,差异有统计学意义(P<0.05);干预组细胞可见空泡变性与凋亡减少,AngⅡ,AT1R,IL?1β,IL?6和TNF?变表达量下降,差异有统计学意义(P<0.05)。结论:氯沙坦可能通过抑制AngⅡ/AT1R的表达,进而减轻百草枯诱导人肾小管上皮细胞的炎症和凋亡。

Objective:To investigate the mechanism and effect of losartan in reducing HK2 injury in the acute paraquat poisoning model.Methods:HK2 were cultured in vitro and divided into the control group,experimental group and intervention group.Half effective doses of paraquat and losartan were measured by CCK8.The control group was in normal medium.The experimental group was added paraquat for 24 hours.The intervention group was added paraquat+losartan for 24 hours.HK2 pathology was made with HE and TUNEL.The mRNA and total protein of the 3 groups of HK2 were extracted and the AngⅡ,AT1R,IL?1β,IL?6 and TNF?αwere measured by RT?PCR,and AT1R was detected by western blot.Results:Compared with the control group,the experimental group showed increased vacuolar degeneration and apoptosis under cystoscopy,while the expression of AngⅡ,AT1R,IL?1β,IL?6 and TNF?αincreased and the differences were statistically significant(P<0.05).Compared with the experimental group,the intervention group showed that vacuole degeneration and apoptosis were reduced under cystoscopy,and the expressions of AngⅡ,AT1R,IL?1β,IL?6 and TNF?αdecreased and the differences were statistically significant(P<0.05).Conclusion:Losartan can reduce the inflammation and apoptosis of HK2 in the acute paraquat poisoning model,which may be by the mechanism of inhibiting the expression of AngⅡ/AT1R.