布托啡诺联合氟比洛芬酯预处理通过ANG(1-7)对小鼠肢体缺血再灌注诱发肾损伤的保护作用及机制

Protective effect and mechanism of butorphanol combined with flurbiprofen axetil preconditioning on renal injury induced by limb ischemia-reperfusion in mice through ANG(1-7)

目的探究布托啡诺联合氟比洛芬酯预处理通过血管紧张素(1-7)[Angiotensin(1-7),ANG(1-7)]对小鼠肢体缺血再灌注诱发肾损伤的保护作用及对炎症通路TNF超家族成员14配体(TNF superfamily member 14 ligand,LIGHT)/TNF超家族成员14(TNF superfamily member 14,HVEM)的影响。方法将45只SPF级BABL/c小鼠随机分为对照组(n=15)、模型组(n=15)、试验组(n=15),除对照组外,建立肢体缺血再灌注诱发肾损伤模型,试验组造模前采用布托啡诺联合氟比洛芬酯预处理12 h,对照组和模型组给予等量生理盐水。HE染色确认各组小鼠肾脏组织病理水平及病理评分,大生化分析确定肾功能指标血清肌酐(serum creatinine,Scr)和尿素氮(urea nitrogen,BUN)水平,检测外周血及肾组织ANG(1-7)表达水平,qPCR和Western blot检测肾脏组织LIGHT和HVEM mRNA和蛋白表达水平。结果与对照组比较,模型组小鼠肾组织损伤升高,病理评分升高,Scr和BUN表达升高,ANG(1-7)表达水平降低,LIGHT和HVEM表达水平升高;与模型组比较,试验组小鼠模型组小鼠肾组织损伤降低,病理评分减低,Scr和BUN表达降低,ANG(1-7)表达水平升高,LIGHT和HVEM表达水平降低(P<0.05)。结论布托啡诺联合氟比洛芬酯预处理对小鼠肢体缺血再灌注诱发肾损伤具有保护作用,其机制可能通过影响ANG(1-7)干预炎症通路LIGHT/HVEM信号蛋白表达相关。

Objective To investigate the protective effect of butorphanol combined with flurbiprofen axetil preconditioning on renal injury induced by limb ischemia-reperfusion in mice through angiotensin(1-7)[ANG(1-7)]and its effect on TNF superfamily member 14 ligand(LIGHT)/TNF superfamily member 14(HVEM)in the inflammatory pathway.Methods Forty-five SPF grade BABL/c mice were randomly divided into control group(n=15),model group(n=15),and experimental group(n=15).Except the control group,renal injury model induced by limb ischemia-reperfusion was established.The experimental group was pretreated with butorphanol and flurbiprofen axetil for 12 h before modeling,and the control group and model group were given the same amount of normal saline.HE staining was used to confirm the pathological level and pathological score of kidney tissues of mice in each group.Biochemical analysis was used to determine the levels of serum creatinine(Scr)and urea nitrogen(BUN),and the levels of ANG(1-7)expression in peripheral blood and kidney tissues.The mRNA and protein expression levels of LIGHT and HVEM in kidney tissues were detected by qPCR and Western blot.Results Compared with the control group,the renal tissue injury,pathological score,and the expression of Scr and BUN decreased,the expression level of ANG(1-7)decreased,and LIGHT and HVEM increased in the model group(P<0.01).Compared with the model group,the renal tissue injury,pathological score,and the expression of Scr and BUN decreased,the expression level of ANG(1-7)increased,and LIGHT and HVEM decreased(P<0.01).Conclusion Butorphanol combined with flurbiprofen axetil preconditioning has a protective effect on renal injury induced by limb ischemia-reperfusion in mice,and its mechanism may be related to the intervention of ANG(1-7)on the expression of LIGHT/HVEM signal pathway proteins.