体外细胞中TWEAK基因介导的TWEAK/Fn14及NF-κB通路关键因子与肌少症的相关性  

Correlation between the expression difference of P53-TWEAK/Fn14-NF-κB pathway mediated by TWEAK gene and sarcopenia

作　　者：王安彦 刘金玲 卓娅·买买提乌斯满[2] 王红梅[2] WANG Anyan;LIU Jinling;Zhuoya Maimaitiwusiman;WANG Hongmei(Graduate School,Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi830001,China;the Second Ward of General Health Internal Medicine,People’s Hospital of Xinjiang Uygur Autonomous Region,Xinjiang Uygur Autonomous Region,Urumqi830001,China)

机构地区：[1]新疆医科大学研究生学院,新疆乌鲁木齐830001 [2]新疆维吾尔自治区人民医院综合保健内科二病区,新疆乌鲁木齐830001

出　　处：《中国医药导报》2025年第5期1-7,46,共8页China Medical Herald

基　　金：国家自然科学基金资助项目(82160276);新疆维吾尔自治区自然科学基金项目(2022D01C127)。

摘　　要：目的探讨肿瘤坏死因子样细胞凋亡弱诱导剂基因介导的TWEAK/Fn14及核因子κB(NF-κB)通路关键因子表达差异与肌少症的相关性。方法实验分为空白对照组、肌少症细胞组、肌少症细胞+阴性对照组、肌少症细胞+TWEAK-shRNA组。地塞米松诱导小鼠C2C12细胞为肌少症细胞模型,免疫荧光观察肌管形成相关蛋白MyHC蛋白的表达;肌少症模型构建成功后转染携带EGFP基因、TWEAK-shRNA基因的慢病毒建立肌少症细胞+阴性对照组、肌少症细胞+TWEAK-shRNA组。qRT-PCR、Western blot法检测TWEAK、Fn14的表达,Western blot法检测p53、IKKα、IKKβ、NF-κB p65、p-p65表达量。预测p53与TWEAK启动子区域结合。结果肌少症细胞组肌球蛋白重链MyHC蛋白表达水平低于空白对照组(P<0.05)。肌少症细胞组细胞肌管数量少于空白对照组(P<0.05);肌少症细胞+TWEAK-shRNA组肌管数量多于肌少症细胞组(P<0.05)。与空白对照组比较,肌少症细胞组TWEAK、Fn14 mRNA及蛋白表达水平升高(P<0.05);肌少症细胞+TWEAK-shRNA组TWEAK、Fn14 mRNA及蛋白表达水平低于肌少症细胞组与肌少症细胞+阴性对照组(P<0.05)。与空白对照组比较,肌少症细胞组p53、IKKα、IKKβ、p-p65蛋白表达水平升高(P<0.05);与肌少症细胞组、肌少症细胞+阴性对照组比较,肌少症细胞TWEAK-shRNA组p53、IKKα、IKKβ、p-p65蛋白表达水平降低(P<0.05)。p53与TWEAK启动子区域结合。结论TWEAK基因通过调节p53、TWEAK/Fn14及下游NF-κB通路关键因子影响肌少症的发生。Objective To investigate the relationship between the expression difference of TWEAK/Fn14 and key factor of nuclear factorκB(NF-κB)pathway mediated by weak inducer of tumor necrosis factor-like apoptosis gene and sarcopenia.Methods The experiment was divided into blank control group,sarcopenia cell group,sarcopenia cell+negative control group,and sarcopenia cell+TWEAK-shRNA group.Dexamethasone-induced mouse C2C12 cells were used as a cell model of sarcopenia,and the expression of myotube formation-related protein MyHC protein was observed by immunofluorescence.After successful construction of sarcopenia model,lentivirus carrying EGFP gene and TWEAK-shRNA gene was transfected to establish sarcopenia cells+negative control group and sarcopenia cells+TWEAK-shRNA group.The expression of TWEAK and Fn14 was detected by qRT-PCR and Western blot.The expression of p53,IKKα,IKKβ,NF-κB p65 and p-p65 was detected by Western blot.The binding of p53 to TWEAK promoter region was predicted.Results The expression level of myosin heavy chain MyHC protein in sarcopenia cell group was lower than that in blank control group(P<0.05).The number of myotubes in the sarcopenia cell group was less than that in the blank control group(P<0.05).The number of myotubes in the sarcopenia cell+TWEAK-shRNA group was more than that in the sarcopenia cell group(P<0.05).Compared with the blank control group,the mRNA and protein expression levels of TWEAK and Fn14 in the sarcopenia cell group were increased(P<0.05).The mRNA and protein expression levels of TWEAK and Fn14 in the sarcopenia cells+TWEAK-shRNA group were lower than those in the sarcopenia cells group and the sarcopenia cell+negative control group(P<0.05).Compared with the blank control group,the expression levels of p53,IKKα,IKKβ,and p-p65 protein in the sarcopenia cell group were increased(P<0.05).Compared with the sarcopenia cell group and the sarcopenia cell+negative control group,the expression levels of p53,IKKα,IKKβand p-p65 protein in the TWEAK-shRNA group were decreased(P

关 键 词：肌少症 TWEAK Fn14 核因子ΚB 小鼠 C2C12 

分 类 号：R-3[医药卫生]