强骨通痹胶囊对大鼠骨关节炎氧化应激的影响及作用机制

The effect and mechanism of Qianggu Tongbi capsules on oxidative stress in rats with osteoarthritis

目的:探讨强骨通痹胶囊对大鼠骨关节炎氧化应激的影响及作用机制。方法:将24只SD大鼠随机分为2组,其中6只作为对照组,18只进行建模;建模完把18只大鼠随机分为模型组、强骨通痹胶囊组和葡萄糖胺组,并对应用药处理;苏木精-伊红染色法(hematoxylin-eosinstaining,HE)观察软骨组织变化;超氧化物歧化酶(superoxide dismutase,SOD)测定试剂盒,微量丙二醛(malondialdehyde,MDA)测定试剂盒和一氧化氮(nitric oxide,NO)测定试剂盒检测血清中SOD,MDA和NO水平;实时聚合酶链式反应(real time-polymerase chain reaction,RT-PCR)检测大鼠关节炎软骨组织中核因子NF-E2相关因子(nuclear factorerythroid 2-relaxed factor 2,Nrf2)、Kelch样ECH相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)、巨噬细胞激活因子(macrophage activation factor,Maf)、血红素氧合酶-1(heme oxygenase-1,HO-1)和醌氧化还原酶1(NADPH quinone oxidoreductase 1,NQO1)mRNA表达水平;Western blot法检测Nrf2,Keap1,Maf,HO-1和NQO1的蛋白表达水平。结果:HE染色结果表明,与空白对照组相比,模型组的软骨组织中有明显的致密炎性细胞浸润,而强骨通痹胶囊组和葡萄糖胺组的炎症细胞明显减少;和模型组相比,强骨通痹胶囊组和葡萄糖胺组的抗氧化指标浓度显著上升,自由基浓度显著下降(P<0.05);和模型组相比,强骨通痹胶囊组和葡萄糖胺组的Nrf2,Maf,HO-1和NQO1的mRNA和蛋白表达水平都有明显上调(P<0.05),而Keap1的mRNA和蛋白表达水平下调。结论:强骨通痹胶囊能通过激活被负调控的Nrf2-Keap1/抗氧化反应元件(antioxidant responsive element,ARE)通路的相关蛋白及其mRNA的表达来改变血清中的抗氧化指标,从而恢复大鼠骨关节炎抗氧化应激的功能。

Objective:To investigate the effect and mechanism of Qianggu Tongbi capsules on oxidative stress in rats with osteoarthritis.Methods:Twenty-four SD rats were randomly divided into two groups,6 of which were used for control group and 18 were used for modeling.After modeling,the 18 rats were randomly divided into model group,Qianggutongbi capsules group,and glucosamine group.Hematoxylin eosin staining(HE) was used to observe the changes of cartilage.Superoxide dismutase(SOD) kit,malondialdehyde(MDA) kit,and nitric oxide(NO) kit were applied to detect the levels of SOD,MDA and NO in serum,respectively.Real time-polymerase chain reaction(RT-PCR) was applied to detect the expressions of nuclear factorerythroid 2-relaxed factor(Nrf2),Kelch-like ECH-associated protein 1(Keap1),macrophage activation factor(Maf),heme oxygenase-1(HO-1),and NADPH quinone oxidoreductase 1(NQO1) in each group.Western blot was used to detect the expressions of Nrf2,Keap1,Maf,HO-1,and NQO1 in each group.Results:HE staining results showed that there was obvious infiltration of dense inflammatory cells in the cartilage tissue of the model group,while the inflammatory cells in the Qianggu Tongbi capsules group and glucosamine group were significantly reduced compared with the blank control group.Compared with the model group,the levels of antioxidant indexes of Qianggu Tongbi capsules group and glucosamine group increased significantly,while the free radical concentrations decreased significantly(P<0.05).Compared with the model group,the mRNA and protein expression levels of Nrf2,Maf,HO-1,and NQO1 in Qianggu Tongbi capsules group and glucosamine group were significantly up-regulated(P<0.05),while the mRNA and protein expression levels of Keap1 were down-regulated(P<0.05).Conclusion:Qianggu Tongbi capsules can change the antioxidant indexes in serum by activating the expression of Nrf2-Keap1/ARE pathway-related proteins and mRNA,so as to restore the function of antioxidant stress in rats with osteoarthritis.