黄芪甲苷通过GATA4途径调控内质网应激对激素性股骨头缺血坏死大鼠的作用机制  

Effect of astragaloside on endoplasmic reticulum stress in rats with steroid-induced avascular necrosis of femoral head (SANFH) through zinc finger transcription factor (GATA 4) pathway

作　　者：李鹏飞 刘宏鹏 李琳琳 曹童 王春龙[4] 王艳 LI Pengfei;LIU Hongpeng;LI Linlin;CAO Tong;WANG Chunlong;WANG Yan(Graduate School of Heilongjiang University of Chinese Medicine,Harbin 150040,China;Department of Bone Injury,the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China;Department of Cardiology,the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China;Department of Bone and Traumatology,the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China;Rehabilitation Center,the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China)

机构地区：[1]黑龙江中医药大学研究生院,哈尔滨150040 [2]黑龙江中医药大学附属第二医院骨伤一科,哈尔滨150001 [3]黑龙江中医药大学附属第二医院心血管二科,哈尔滨150001 [4]黑龙江中医药大学附属第二医院骨伤二科,哈尔滨150001 [5]黑龙江中医药大学附属第二医院康复中心,哈尔滨150001

出　　处：《新疆医科大学学报》2024年第10期1340-1347,共8页Journal of Xinjiang Medical University

基　　金：黑龙江省中医药科研项目(ZHY2022-103);黑龙江中医药大学附属第二医院孙申田青年人才基金项目(2021KY-09)。

摘　　要：目的探讨黄芪甲苷(As-Ⅳ)通过锌指转录因子(GATA4)途径调控内质网应激对激素性股骨头缺血坏死(SANFH)大鼠的影响及其作用机制。方法58只SD大鼠,随机选取10只作为对照组(皮下注射生理盐水,1周7次,连续4周),其余48只大鼠复制SANFH模型,8只未建模成功,40只大鼠建模成功后,随机分为模型组、GATA 4干扰组、As-Ⅳ组、As-Ⅳ+GATA 4干扰组,每组10只。造模4周开始As-Ⅳ组、As-Ⅳ+GATA 4干扰组分别灌胃40 mg/kg的As-Ⅳ,余灌胃等量生理盐水,1次/d,共20 d;GATA4干扰组、As-Ⅳ+GATA4干扰组于造模第1周开始尾静脉注射GATA4干扰载体,1次/周,共4次。比较各组股骨Micro-CT扫描参数、病理学变化、血清氧化应激水平及股骨头组织中GATA4、丙酮酸激酶M2型(PKM2)及骨代谢相关核因子κB受体激活因子(RANK)、RANK配体(RANKL)及内质网应激相关蛋白CAAT增强子结合蛋白同源蛋白(CHOP)、葡萄糖调节蛋白78(GRP78)、半胱氨酸蛋白水解酶12(Caspase-12)蛋白表达情况。结果与GATA 4干扰组比较,As-Ⅳ+GATA 4干扰组、As-Ⅳ组的骨小梁数量、骨小梁厚度、骨体积分数依次升高,骨小梁分离度依次降低(P<0.05)。HE染色显示,模型组大鼠股骨头出现严重坏死,GATA 4干扰组股骨头病变情况加重,As-Ⅳ+GATA 4干扰组、As-Ⅳ组大鼠股骨头坏死情况减轻。与GATA 4干扰组比较,As-Ⅳ+GATA 4干扰组、As-Ⅳ组血清超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)水平、GATA 4、PKM2蛋白相对表达量依次升高,血清丙二醛(MDA)水平、RANKL、RANK、CHOP、GRP78、Caspase-12蛋白相对表达量依次降低(P<0.05)。结论As-Ⅳ有助于缓解SANFH大鼠股骨头坏死进展,其机制可能与激活GATA 4/PKM2通路抑制内质网应激有关。Objective To investigate the effect of AstragalosideⅣ(As-Ⅳ)on endoplasmic reticulum stress in rats induced by hormone-induced ischemic necrosis of femoral head(SANFH)through zinc finger transcription factor(GATA4)pathway and its mechanism.Methods 10 of 58 SD rats were randomly selected as control group(subcutaneous injection of normal saline,7 times a week,for 4 weeks).The remaining 48 rats replicated SANFH model,8 rats failed to establish the model,and 40 rats were randomly divided into model group,GATA4 interference group,As-Ⅳgroup and As-Ⅳ+GATA4 interference group,with 10 rats in each group.4 weeks after modeling,As-Ⅳgroup and As-Ⅳ+GATA4 interference group were given 40 mg/kg of As-Ⅳ,respectively,and the other groups were given the same amount of normal saline once a day for a total of 20 days.GATA4 interference group and As-Ⅳ+GATA4 interference group were injected with GATA4 interference carrier in the tail vein from the first week of modeling,once a week,a total of 4 times.The Micro-CT scanning parameters,pathological changes,serum oxidative stress levels and GATA4,pyruvate kinase M2 type(PKM2),bone metabolisation-related nuclear factorκB receptor activator(RANK),RANK ligand(RANKL),endoplasmic reticulum stress related protein CAAT enhancer binding protein homologous protein(CHOP),78-kDa glucose-regulated protein(GRP78),cysteine proteolytic enzyme 12(Caspase-12)in femoral head tissue were compared.Results Compared with GATA4 interference group,bone trabecular number,bone trabecular thickness and bone volume fraction in As-Ⅳ+GATA4 interference group and As-Ⅳinterference group were increased successively,while bone trabecular separation degree was decreased successively(P<0.05).HE staining showed that there was severe necrosis of the femoral head in the model group,the lesions of the femoral head in GATA4 interference group were aggravated,and the necrosis of the femoral head in As-Ⅳ+GATA4 interference group and As-Ⅳgroup was alleviated.Compared with GATA4 interference group,the levels

关 键 词：激素性股骨头缺血坏死 黄芪甲苷 锌指转录因子 内质网应激 机制 

分 类 号：R681.8[医药卫生—骨科学]