汉黄芩苷对肾缺血/再灌注损伤大鼠肾功能及NF-κB/iNOS/NO通路的影响

Influences of Wogonoside on Renal Function and NF-κB/iNOS/NO Pathway in Rats with Renal Ischemia/Reperfusion Injury

目的观察汉黄芩苷对肾缺血/再灌注(I/R)损伤大鼠肾功能及对NF-κB/iNOS/NO通路的影响并探讨其机制。方法制备肾I/R模型大鼠,按随机数字表法分为模型组、乌司他丁组及汉黄芩苷低(5 mg/kg)、中(25 mg/kg)、高(75 mg/kg)剂量组,模型组及假手术组给予等量0.9%氯化钠注射液灌胃。再灌注后24 h,检测各组大鼠肾功能,苏木素-伊红(HE)染色观察肾组织病理学变化,试剂盒检测肾组织中过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、一氧化氮合酶(iNOS)活性、还原性谷胱甘肽(GSH)、丙二醛(MDA)含量及一氧化氮(NO)水平,蛋白印迹(WB)法检测肾组织中NF-κB表达情况。结果与假手术组比较,模型组大鼠肾功能显著下降,血液中血清肌酐(Scr)、血尿素氮(BUN)含量显著升高,肾组织出现肾小管上皮细胞水肿、脱落、小管腔坏死等病理变化,组织病理评分明显升高,CAT、SOD、GSH活性显著下降,MDA含量显著升高,NF-κb蛋白核转移水平明显增加,iNOS活性、NO水平显著增高;与模型组比较,汉黄芩苷低、中、高剂量组大鼠肾功能均明显恢复,肾组织病理变化均显著改善,组织病理评分显著降低,CAT、SOD、GSH活性显著升高,MDA含量显著下降,NF-κb蛋白核转移水平明显减少,iNOS活性、NO水平显著降低,均呈剂量依赖效应(均P<0.05)。结论汉黄芩苷具有提高肾功能,保护缺血/再灌注所致肾损伤的功能,其机制可能是通过抑制氧化应激反应和NF-κB/iNOS/NO通路的激活而实现。

Objective： To investigate the influences of Wogonoside on renal function and NF-κB/iNOS/NO pathway in rats with renal ischemia/reperfusion（I/R） injury. Methods： The rats of renal I/R model were prepared and randomly divided into the model group,Ulinastatin group,Wogonoside low（5 mg/kg）,middle（25 mg/kg）,and high（75 mg/kg） dose groups. The model group and the sham operation group were given the same amount of normal saline by intragastric administration. 24 h after reperfusion,the renal function of rats in each group was detected;the pathological changes of renal tissue were observed by hematoxylin eosin（HE） staining;kit was used to detect the activities of catalase（CAT）,superoxide dismutase（SOD）,inducible nitric oxide synthase（iNOS）,the contents of reduced glutathione（GSH）,malondialdehyde（MDA） content and nitric oxide（NO） level in renal tissue,the expression of NF-κB in renal tissue was detected by Western blot（WB）. Results： Compared with the sham operation group,the renal function of the rats in the model group decreased significantly;the contents of serum creatinine（Scr） and blood urea nitrogen（BUN） in blood increased significantly;the pathological changes such as edema,abscission,necrosis of renal tubular epithelial cells in the renal tissues were observed;histopathological score was significantly increased;the activities of CAT,SOD and GSH decreased significantly,and the content of MDA increased significantly;the nuclear transfer level of NF-κB protein increased significantly;the activity of iNOS and the level of NO increased significantly. Compared with the model group,the renal function of the rats in the low,middle and high dose groups of Wogonoside was obviously restored;the pathological changes of renal tissue were significantly improved;histopathological score was significantly reduced;the activities of CAT,SOD and GSH increased significantly;the content of MDA decreased significantly;the nuclear transfer level of NF-κB protein decreased significantly;the activity of iNOS and the level of NO decreased significantly,and all showed a dose dependent effect. Conclusion： Wogonoside can enhance renal function and protect renal injury induced by ischemia/reperfusion,its mechanism may be achieved by inhibiting oxidative stress reaction and activating NF-κB/iNOS/NO pathway.