【作者】 赵峰；

【导师】 吴帅；

【作者基本信息】 青岛大学 ， 外科学（泌尿外科）（专业学位）， 2020， 硕士

【摘要】 目的:探讨肾气丸在小鼠I/R诱导的急性肾损伤中的作用及机制,为该药物在临床的应用提供参考依据。方法:健康雄性C57BL6小鼠40只,利用随机数字表法分为4组(每组10只):(1)假手术组(Sham);(2)肾缺血再灌注组(肾I/R组);(3)肾气丸低剂量组(10.5g/kg/d);(4)肾气丸高剂量组(21.0g/kg/d)。分组后肾气丸低剂量组给予10.5g/kg/d的肾气丸灌胃,肾气丸高剂量组给予21.0g/kg/d的肾气丸灌胃,持续2周;假手术组和肾I/R组每天给予等量生理盐水灌胃。2周后构建肾I/R损伤模型,24h处死小鼠并取材。检测血清肌酐(Scr)、尿素氮(BUN)及肾脏组织中丙二醛(MDA)含量和超氧化物歧化酶(SOD)的活性;HE染色及TUNEL染色检测肾组织损伤及凋亡水平;PCR及免疫组化检测Bcl-2、Caspase-3表达水平;Western blot法检测PI3K和AKT蛋白表达水平。结果:HE及TUNEL染色表明,肾气丸可抑制细胞损伤及凋亡。与假手术组比较,肾I/R组Scr、BUN、MDA含量、Caspase-3水平均升高(P<0.05),SOD活性、Bcl-2水平、PI3K和AKT蛋白表达明显下降(P<0.05);与肾I/R组比较,肾气丸组(低剂量组和高剂量组)Scr、BUN和MDA含量、Caspase-3水平均降低(P<0.05),而SOD活性、Bcl-2水平、PI3K和AKT蛋白表达升高(P<0.05);与肾气丸低剂量组比较,肾气丸组高剂量组Scr、BUN和MDA含量、Caspase-3水平均降低(P<0.05),SOD活性、Bcl-2水平、PI3K和AKT蛋白表达均升高(P<0.05)。结论:肾气丸可通过激活PI3K/AKT信号通路对小鼠I/R诱导的急性肾损伤起保护作用。更多还原

【Abstract】 Objective:To investigate the protective effect and mechanism of Shen-Qi pills on acute renal injury induced by I/R in mice.Methods:The total of 40 healthy male C57BL6 mice were randomly divided into 4 groups:(1)sham group;(2)renal ischemia reperfusiongroup(renal I/R group);(3)low dose group of Shen-Qi pills;(4)high dose group of Shen-Qi pills.The low dose group of Shen-Qi pills were administered 10.5 mg/kg/d of Shen-Qi pills,while the high dose group were gavaged by 21.0 mg/kg/d,for two weeks.The sham and I/R groups were administered the same amount of normal saline every day.After 2 weeks,the model of renal I/R injury was duplicated.The 4 groups were sacrificed after 24 h.Then Scr,BUN,the content of MDA and activity of SOD weredetected.HE and TUNEL staining were used to detect renal injury and apoptosis level.Andthe expression levels of bcl-2 and caspase-3 were detected by PCR and Immunohistochemistry.The expression levels of PI3 K and AKT were detected by Western blot.Results:HE and TUNEL staining showed that the Shen-Qi pills could inhibit cell damageand apoptosis.Compared with the sham group,Scr,BUN,MDA and caspase-3 levelsinthe renal I/R group were all increased(P<0.05),and SOD activity,bcl-2 level,PI3 Kand AKT protein expression level were significantly decreased(P<0.05).Compared with therenal I/R group,Scr,BUN,MDA and caspase-3 level in the Shen-Qi pills group(low dose group and high dose group)decreased(P<0.05),while SOD activity,bcl-2 level,PI3 K and AKT protein expression level were increased(P<0.05).Compared with thelow-dose group,the content of Scr,BUN and MDA and the level of caspase-3 in the high-dose group were all decreased(P<0.05),while the SOD activity,bcl-2 level,PI3 K and AKT protein expression level were all increased(P<0.05).Conclusion:The Shen-Qi pills can protect mice from I/R induced by acute renal injury by activating the PI3K/AKT signaling pathway.更多还原