基于Keap1/Nrf2信号通路探讨苗药痛风方对痛风性关节炎大鼠的作用机制  

Effect of Miaoyao Tongfeng prescription on Keap1/Nrf2 signaling pathway in gouty arthritis rats

作　　者：刘道忠 冯佳 刘义 熊国飞 张宗星 江露 李玮怡 包卓玛 袁林 LIU Daozhong;FENG Jia;LIU Yi;XIONG Guofei;ZHANG Zongxing;JIANGLu;LI Weiyi;BAO Zhuoma;YUAN Lin(Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi 445000,China;Xianfeng County Hospital of Traditional Chinese Medicine,Xianfeng 445600,China;Hubei Provincial Clinical Medical Research Center for Nephropathy,Enshi 445000,China)

机构地区：[1]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000 [2]咸丰县中医医院,湖北咸丰445600 [3]湖北省肾脏病临床医学研究中心,湖北恩施445000

出　　处：《中国病理生理杂志》2024年第8期1505-1510,共6页Chinese Journal of Pathophysiology

基　　金：湖北省教育厅项目(No.B2019089);湖北民族大学2023年研究生教育创新项目(No.MYK2023063)。

摘　　要：目的:探讨苗药痛风方(MTP)是否通过调控Kelch样环氧氯丙烷相关蛋白1(Keap1)/核因子E2相关因子2(Nrf2)信号通路,减轻痛风性关节炎模型大鼠氧化应激损伤。方法:将50只雄性SD大鼠随机分为对照组、模型组、MTP组、Nrf2抑制剂全反式维甲酸(ATRA)组和MTP组+ATRA组。使用3%氧嗪酸钾溶液(10 mL/kg)行腹腔内注射,每天2次,连续注射1周,然后在大鼠右侧膝关节注射0.2 mL尿酸钠混悬液(25 g/L),构建痛风性关节炎模型。各组大鼠于造模后48 h处死取材,测量大鼠膝关节肿胀程度;HE染色观察大鼠膝关节病理改变;试剂盒检测大鼠血清中超氧化物歧化酶(SOD)、丙二醛(MDA)和白细胞介素1β(IL-1β)水平;qRT-PCR法检测大鼠膝关节滑膜中Nrf2、Keap1、血红素加氧酶1(HO-1)、IL-1β和NAD(P)H:醌氧化还原酶1(NQO1)的mRNA表达;Western blot检测大鼠膝关节滑膜中Nrf2、Keap1和NQO1的蛋白表达。结果:与模型组相比,MTP能显著降低大鼠膝关节肿胀程度,减轻膝关节滑膜的病理损伤,抑制MDA和IL-1β的表达,增加SOD的表达,上调滑膜中HO-1 mRNA及Nrf2和NQO1 mRNA和蛋白的表达,下调IL-1βmRNA及Keap1 mRNA和蛋白的表达。结论:MTP通过调控Nrf2及其下游抗氧化基因发挥防治大鼠痛风性关节炎的作用。AIM:To investigate whether Miaoyao Tongfeng prescription(MTP)attenuates oxidative stress injury in gouty arthritis rats by regulating Kelch-like epicklorohydrin-related protein 1(Keap1)/nuclear factor E2-related factor 2(Nrf2)signaling pathway.METHODS:Fifty male Sprague-Dawley rats were randomly divided into control group,model group,MTP group,Nrf2 inhibitor all-trans retinoic acid(ATRA)group and MTP+ATRA group.The 3% potassium oxonate solution(10 mL/kg)was injected intraperitoneally twice a day for 1 week,and then 0.2 mL sodium urate suspension(25 g/L)was injected into the right knee joints to establish the gouty arthritis model.All rats were sacrificed 48 h after modeling.Grades of swelling in the knee joints were evaluated,and HE staining was used to observe the pathological changes of knee joints.Serum levels of superoxide dismutase(SOD),malondialdehyde(MDA)and interleukin-1β(IL-1β)were detected using kits.The mRNA levels of Nrf2,Keap1,heme oxygenase-1(HO-1),IL-1β,NAD(P)H:quinone oxidoreductase 1(NQO1)in the synovial membrane of the knee joints were detected by qRT-PCR.Western blot was used to detect the expression of Nrf2,Keap1 and NQO1 in the synovial membrane of the knee joints.RESULTS:Compared with model group,MTP significantly reduced the degree of knee joint swelling,and attenuated the pathological injury of knee synovial membrane.It also inhibited the expression of MDA and IL-1β,but increased the expression of SOD.The mRNA expression of HO-1,and the mRNA and protein expression of Nrf2 and NQO1 in synovium were up-regulated,while the levels of IL-1βmRNA,Keap1 mRNA and Keap1 protein were down-regulated.CONCLUSION:Miaoyao Tongfeng prescription shows promise in the prevention and treatment of gouty arthritis through regulating Nrf2 and downstream anti-oxidation genes.

关 键 词：苗药痛风方 痛风性关节炎 Keap1/Nrf2信号通路 

分 类 号：R285.5[医药卫生—中药学] R684.3[医药卫生—中医学] R363