金丝桃苷调控Nrf2/HO-1/NQO1通路减轻大鼠肾缺血再灌注损伤

Hyperoside regulates Nrf2/HO-1/NQO1 signaling pathway to alleviate renal ischemiareperfusion injury in rats

目的探究金丝桃苷对大鼠肾缺血再灌注(IR)损伤及核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)/醌氧化还原酶1(NQO1)信号通路的调节。方法以结扎双肾动脉60 min再恢复灌注120 min构建肾IR大鼠模型,将40只SD大鼠随机均分为对照组,模型组,金丝桃苷12.5、25、50 mg·kg^(-1)组,留取大鼠血液和肾脏组织标本,检测24 h蛋白尿、血肌酐及尿素氮、超氧化物歧化酶(SOD)和丙二醛(MDA)水平,HE染色观察肾脏组织病变,免疫组化法检测caspase-3表达,qRT-PCR法检测诱导型一氧化氮合酶(iNOS)、白细胞介素(IL)-6、IL-10 mRNA表达,Western blot法检测肾组织中Bcl-2、Bax、Nrf2、HO-1、NQO1蛋白表达。另取32只大鼠随机均分为对照组、模型组、金丝桃苷50 mg·kg^(-1)组和金丝桃苷+ML385(30 mg·kg^(-1))组,进行上述检测。结果与模型组比较,金丝桃苷25、50 mg·kg^(-1)组24 h蛋白尿、血肌酐、尿素氮、MDA水平均显著下降,SOD水平显著升高(P<0.05)。模型组肾组织出现明显病理损伤,炎症细胞浸润严重,金丝桃苷组肾组织病理损伤均减轻,50 mg·kg^(-1)组改善更为明显。与模型组比较,金丝桃苷25、50 mg·kg^(-1)组Bax/Bcl-2、caspase-3蛋白表达和iNOS、IL-6 mRNA表达显著下降,IL-10 mRNA和p-Nrf2/Nrf2、HO-1及NQO1蛋白表达显著上调(P<0.05),金丝桃苷12.5 mg·kg^(-1)组Bax/Bcl-2表达显著下降(P<0.05)。ML385可部分阻断金丝桃苷对肾IR大鼠的改善作用。结论金丝桃苷可能通过上调Nrf2/HO-1/NQO1信号通路,抑制肾脏细胞的凋亡、过度的氧化应激和炎症反应,减轻肾脏IR损伤。

AIM To explore the regulation effect of hyperoside on renal ischemia-reperfusion(IR)injury and nuclear factor E2 related factor 2(Nrf2)/heme oxygenase 1(HO-1)/quinone oxidoreductase 1(NQO1)signaling pathway in rats.METHODS The models of renal IR rat were constructed by ligating bilateral renal arteries(60 minutes)and recovering perfusion(120 minutes).Forty SD rats were randomly divided into control group,model group and hyperoside 12.5,25,50 mg·kg^(-1)groups.The samples of blood and renal tissues were collected to detect levels of 24 h proteinuria,serum creatinine,blood urea nitrogen and superoxide dismutase(SOD),malondialdehyde(MDA).The renal tissue lesions were observed by HE staining.The expression of caspase-3 was detected by immunohistochemistry.The expression of inducible nitric oxide synthase(iNOS),interleukin(IL)-6 and IL-1O mRNA was detected by qRT-PCR.The expression of Bcl-2,Bax,Nrf2,HO-1 and NQO1 proteins in renal tissues was detected by Western blot.A total of 32 rats were randomly and averagely divided into control group,model group,hyperoside(50 mg·kg^(-1))group and hyperoside+ML385(30 mg·kg^(-1))group for relevant detection.RESULTSSCompared with the model group,the levels of 24 h proteinuria,serum creatinine and blood urea nitrogen,and MDA were significantly decreased,while the levels of SOD were significantly increased in the hyperoside 25 and 50 mg·kg^(-1)groups(P<0.05).HE staining showed that there was obvious pathological injury of renal tissues and inflammatory cells infiltration was severe in the model group.The pathological injury were relieved in the hyperoside groups,and the improvement were more significant in the hyperoside 50 mg·kg^(-1)group.Compared with the model group,the expression of Bax/Bcl-2,caspase-3,iNOS and IL-6 mRNA were decreased significantly,while the expression of IL-10 mRNA,p-Nrf2/Nrf2,HO-1 and NQO1 and SOD were significantly upregulated in the hyperoside 25 and 50 mg·kg^(-1)groups(P<0.05).The expression of Bax/Bcl-2 decreased significantly in the hyperoside 12.5 mg·kg^(-1)group(P<0.05).ML385 can partially block the improvement effect of hyperoside on renal IR rats.CONCLUSION Hyperoside may inhibit apoptosis of renal cells,excessive oxidative stress and inflammatory response by up-regulating Nrf2/HO-1/NQO1 signaling pathways,and relieve renal IR injury.