三七总皂苷预处理对缺血再灌注损伤大鼠肾脏wnt4蛋白的影响

摘要

目的 探讨三七总皂苷(PNS)预处理对缺血再灌注损伤肾脏wnt4蛋白的影响.方法 取72只SD雄性大鼠随机分为对照(CON)组、缺血再灌注损伤(IR)组、PNS组.实验干预:(1)CON组切除右肾后,左肾暴露30 min后关腹.(2)IR组采用右肾切除,左侧肾动脉夹闭30 min恢复血流方法 造模.(3)PNS组实验前尾静脉注射PNS(150 mg/kg),共3 d,其余处理同IR组.在缺血再灌注后24 h、48 h、72 h,每个时间点每组处死8只大鼠,取血清检测肌酐(Scr)、尿素氮(BUN),留取左肾行HE染色,光镜下观察病理变化并进行肾小管损伤评分,免疫组化检测wnt4在肾组织内的表达情况,蛋白印记检测各组wnt4表达量.结果 IR组可见明显的肾脏病理损伤,肾小管损伤评分、Scr、BUN在24 h达到损伤顶峰,此后逐渐下降,72 h仍高于CON组(P<0.05),而PNS组各时间点肾小管损伤评分、Scr、BUN均较IR组降低(P<0.05).wnt4的免疫组化及蛋白印迹结果 一致,即IR组、PNS组wnt4表达量在24 h达到高峰,但PNS组各时间点表达量均较IR组升高(P<0.05).结论 PNS对缺血再灌注损伤的肾脏具有保护作用,其作用机制可能与其增强wnt4蛋白表达有关.%Objective To observe the effect of panax notoginseng saponins (PNS) pretreatment on the expression of wnt4 protein in renal ischemia reperfusion injury in rats. Methods Seventy-two male SD rats were randomly averaged into control group (CON group), renal ischemia-reperfusion group (IR group) and PNS group. Rats in CON group were removed the right kidneys, and the left kidneys were exposed for 30 minutes, and then the incisions were closed. Rats of IR group were removed the right kidneys, and the left renal artery were clamped for 30 minutes, and then the incisions were closed. Rats in PNS group were injected PNS (150 mg/kg) via tail vein for 3 days before experiment, and the rest steps were the same with IR group. After 24 h, 48 h and 72 h reperfusion, 8 rats in each group were killed, and blood and kidney samples were collected to examine the serum creatinine (Scr) and blood urea nitrogen (BUN). The left renal tissues were stained with HE. Pathological changes were observed under light microscopy. The score of renal tubular injury was evaluated. Immunohistochemistry was performed to detecte the expression of wnt4 in kidney tissue. The expression of wnt4 was detected by Western blot assay. Results There was significant renal injury in IR group. The renal tubular injury score, Scr and BUN reached the peak levels of injury at 24 h, which were then decreased gradually, but at 72 h still significantly higher than those of CON group (P<0.05). The renal tubular injury score, Scr and BUN values at different time points were significantly lower in PNS group than those of IR group (P<0.05). The results of wnt4 immunohistochemistry and Western blot assay were consistent, which showed that the expression of wnt4 peaked at 24 h in IR group and PNS group, but the expression levels were significantly higher in PNS group than those of IR group at each time point (P<0.05). Conclusion PNS could protect kidney from ischemia-reperfusion injury, which may be related to the enhanced expression of wnt4 protein.