刺芒柄花素对顺铂所致大鼠急性肾损伤的保护作用及其机制

Protective effect of formononetin on acute kidney injury induced by cisplatin in rats and its mechanism

目的:探究刺芒柄花素对顺铂所致的急性肾损伤(AKI)大鼠的保护作用及其机制。方法:将32只8周龄雄性SD大鼠随机分为阴性组、顺铂组、刺芒柄花素+顺铂组和刺芒柄花素组。顺铂组及刺芒柄花素+顺铂组大鼠腹腔注射顺铂(20 mg/kg)建立AKI模型。AKI造模成功24 h后,刺芒柄花素+顺铂组和刺芒柄花素组大鼠采用刺芒柄花素(15 mg/kg)灌胃3 d,阴性组与顺铂组给予等量0.9%生理盐水灌胃。最后一次灌胃给药后,取各组大鼠血液以检测肾功能;取肾脏组织行HE染色评估大鼠肾小管损伤程度;RT-qPCR、Western blot检测肾组织中OPA1、Drp1、p66 Shc、Caspase-9、mfn1、CytC的表达水平;并检测各组大鼠肾组织细胞的活性氧(ROS)代谢情况。结果:与阴性组相比,顺铂组大鼠的血尿素与血肌酐水平上升(P<0.05),肾组织出现较明显的病理损伤。与顺铂组相比,顺铂+刺芒柄花素组大鼠OPA1及CytC mRNA相对表达量上升(P<0.05),Drp1、p66 Shc、Caspase-9 mRNA的相对表达量均降低(P<0.05),而mfn1 mRNA的相对表达差异无统计学意义(P>0.05);但OPA1、mfn1、CytC蛋白相对表达量均上升(P<0.05),且Drp1、p66 Shc、Caspase-9蛋白的相对表达量均降低(P<0.05);肾组织细胞ROS的含量下降(P<0.05)。结论:刺芒柄花素对顺铂所致的急性肾损伤大鼠具有一定的保护作用,其机制可能与调节线粒体分裂、抑制ROS的产生和抑制细胞凋亡有关。

Objective:To investigate the protective effect and mechanism of formononetin on acute kidney injury(AKI)induced by cisplatin in rats.Methods:328-week-old male SD rats were randomly divided into negative group,cisplatin group,formononetin+cisplatin group and formononetin group.Rats in cisplatin group and formononetin+cisplatin group were intraperitoneally injected with cisplatin(20 mg/Kg)to establish AKI model.24 h later,formononetin+cisplatin group and formononetin group were given formononetin(15 mg/Kg)intragastrically for 3 days,and the negative group and cisplatin group were given 0.9%normal saline intragastrically.After the last intragastric administration,blood of rats in each group was collected to detect renal function.HE staining was performed to evaluate the degree of renal tubule injury in rats.The expression levels of OPA1,Drp1,P66 Shc,Caspase-9,MFN1 and CytC in renal tissues were detected by RT-QPCR and Western blot.The reactive oxygen species(ROS)metabolism of renal cells in each group was detected.Results:Compared with the negative group,the levels of blood urea and creatinine in cisplatin group were significantly increased(P<0.05),and the renal tissue showed obvious pathological damage.Compared with cisplatin group,the mRNA relative expression levels of OPA1 and CytC in cisplatin+formononetin group were significantly increased(P<0.05),while the mRNA relative expression level of MFN1 was not significantly changed(P<0.05),the relative expression levels of Drp1,P66 Shc and Caspase-9 mRNA were significantly decreased(P<0.05).There was no significant difference in the relative expression of mfn1 mRNA(P>0.05).The relative expression levels of OPA1,MFN1 and CytC were significantly increased(P<0.05),and the relative expression levels of Drp1,P66 Shc and Caspase-9 were significantly decreased(P<0.05).ROS content in renal tissue cells decreased significantly(P<0.05).Conclusion:Formononetin has certain protective effect on acute kidney injury induced by cisplatin in rats,and its mechanism may be related to regulating mitochondrial division,inhibiting ROS production and inhibiting apoptosis.