二甲双胍基于Nrf2/HO-1/NF-κB信号通路对腰椎间盘退变的影响  

Effect of metformin on lumbar disc degeneration based on Nrf2/HO-1/NF-κB signaling pathway

作　　者：董军立[1] 贾诚谦 姚宪宝 苗二芽 蔡毅[1] DONG Junli;JIA Chengqian;YAO Xianbao;MIAO Erya;CAI Yi(Department of Pain Management,the Central Hospital of Wuhan,Tongji Medical College of Huazhong University of Science and Technology,Wuhan 430014,Hubei,China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院疼痛科,湖北省武汉市430014

出　　处：《广西医学》2023年第11期1316-1325,共10页Guangxi Medical Journal

基　　金：湖北省武汉市卫生和计划生育委员会科研项目(WX18Q23)。

摘　　要：目的探讨二甲双胍基于核转录因子红系2相关因子2(Nrf2)/血红素氧合酶1(HO-1)/核转录因子κB(NF-κB)信号通路对腰椎间盘退变的影响。方法选取120只雄性新西兰大白兔,选取其中100只大白兔建立腰椎间盘退变模型,并随机分为模型组(灌胃生理盐水10 mL/kg)、二甲双胍低剂量组(灌胃二甲双胍40 mg/kg)、二甲双胍高剂量组(灌胃二甲双胍160 mg/kg)、Nrf2抑制剂组(腹腔注射全反式维甲酸7 mg/kg)、二甲双胍+Nrf2抑制剂组(灌胃二甲双胍160 mg/kg+腹腔注射全反式维甲酸7 mg/kg),每组20只。将其余20只大白兔作为假手术组(灌胃生理盐水10 mL/kg)。各组大白兔给予相应干预4周后,进行X线及MRI检查,以观察腰椎骨质改变情况及退变程度;采用HE染色法观察各组大白兔椎间盘组织形态变化;采用免疫组化染色法检测各组大白兔椎间盘组织Ⅱ型胶原蛋白、Nrf2阳性表达水平;采用ELISA检测各组大白兔椎间盘组织白细胞介素(IL)-6、IL-1β、活性氧簇、丙二醛水平;采用Western blot检测各组大白兔椎间盘组织Nrf2/HO-1/NF-κB信号通路相关蛋白、超氧化物歧化酶2(SOD2)蛋白、过氧化氢酶(CAT)蛋白、Caspase-3蛋白、Ⅰ型胶原蛋白表达量。结果(1)与假手术组相比,模型组大白兔的椎间盘纤维环及髓核细胞坏死严重,L 4~5椎间盘相对高度降低,椎间盘Prirrmann分级评分升高,椎间盘组织中的Ⅱ型胶原蛋白及Nrf2平均光密度值均降低,IL-6、IL-1β、活性氧簇、丙二醛水平均升高,Nrf2蛋白、HO-1蛋白、SOD2蛋白、CAT蛋白、Ⅰ型胶原蛋白表达量均降低,磷酸化NF-κB(p-NF-κB)/NF-κB值、Caspase-3蛋白表达量均升高(均P<0.05)。(2)与模型组相比,二甲双胍低剂量组、二甲双胍高剂量组大白兔的椎间盘纤维环及髓核细胞坏死情况有所改善,L 4~5椎间盘相对高度均升高,椎间盘Prirrmann分级评分均降低,椎间盘组织中的Ⅱ型胶原蛋白及Nrf2平均光密度值均升高Objective To explore the effect of metformin on lumbar disc degeneration based on nuclear factor-erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/nuclear factor kappa B(NF-κB)signaling pathways.Methods A total of 120 male New Zealand big white rabbits were selected,therein 100 big white rabbits of them were selected to establish the lumbar disc degeneration model,and they were randomly assigned to model group(intragastric administration with 10 mL/kg normal saline),low-dose metformin group(intragastric administration with 40 mg/kg metformin),high-dose metformin group(intragastric administration with 160 mg/kg metformin),Nrf2 inhibitor group(intraperitoneal injection with 7 mg/kg all-trans retinoic acid),or metformin+Nrf2 inhibitor group(intragastric administration with 160 mg/kg metformin+intraperitoneal injection with 7 mg/kg all-trans retinoic acid),with 20 big white rabbits in each group.The remaining 20 big white rabbits were taken as sham operation group(intragastric administration with 10 mL/kg normal saline).After giving a 4-week intervention to big white rabbits of various groups,the X ray and MRI examination were conducted to observe the osseous changes and degeneration degree of lumbar vertebra.The morphological changes of lumbar disc tissues of big white rabbits in various groups were observed by employing the HE staining.The positive expressions of typeⅡcollagen and Nrf2 in intervertebral disc tissues of big white rabbits in various groups were detected by using the immunohistochemical staining.The levels of interleukin(IL)-6,IL-1β,reactive oxygen species,and malondialdehyde in intervertebral disc tissues of big white rabbits in various groups were detected by using the ELISA.The expressions of proteins related to Nrf2/HO-1/NF-κB signaling pathways,superoxide dismutase 2(SOD2)protein,catalase(CAT)protein,Caspase-3 protein,and typeⅠcollagen in intervertebral disc tissues of big white rabbits in various groups were detected by employing the Western blot.Results(1)Compared with the sham o

关 键 词：二甲双胍 腰椎间盘退变 椎间盘组织 核转录因子红系2相关因子2 血红素氧合酶1 核转录因子κB 大白兔 

分 类 号：R681.533[医药卫生—骨科学]