【摘要】 目的探讨TDP-43基因表达干预炎症因子和缺血缺氧应激依赖的JNK和p38MAPK信号通路的机制。方法选取60只SD大鼠,分为正常对照组(正常SD大鼠,不进行任何处理)、骨关节炎(OA)模型组和TDP-43-m MSCs+OA组,每组20只。OA模型组和TDP-43-m MSCs+OA组,采用胶原酶注射建立大鼠OA模型。对TDP-43-m MSCs+OA组大鼠进行TDP-43-m MSCs移植。采用酶联免疫吸附(ELISA)法测定大鼠血清中炎症因子肿瘤坏死因子α(TNF-α)和白介素1β(IL-1β)的水平;采用Western blot法检测各组大鼠软骨组织中TDP-43、RACK1、MTK1、MAPKKK磷酸化的表达、应激颗粒(SGs)的形成及JNK和p38 MAPK信号通路的蛋白变化。结果 OA模型组大鼠软骨组织中TDP-43的蛋白表达显著低于正常对照组,TDP-43-m MSCs+OA组显著高于正常对照组和OA模型组,TDP-43-m MSCs移植成功。与正常对照组相比,OA模型组大鼠软骨组织中胞质SGs的形成量显著降低,TDP-43-m MSCs+OA组显著高于模型组;与正常对照组相比,OA模型组大鼠软骨组织中RACK1、JNK和p38 MAPK的蛋白表达、MTK1、MAPKKK磷酸化的表达量,大鼠血清中TNF-α和IL-1β的分泌量均显著高于正常对照组,而TDP-43-m MSCs+OA组以上指标均显著低于模型组。结论 TDP-43基因的过表达,可以干预炎症因子的分泌,抑制缺血缺氧应激依赖的JNK和p38 MAPK信号通路的激活,从而逆转软骨细胞病变的分子机制。上述结论为OA的治疗提供新的理论依据。更多还原

【Abstract】 Objective To investigate the mechanism of TDP-43 gene expression on inflammatory factors and JNK and p38 MAPK signaling pathways in ischemic hypoxic stress dependence. Methods 60 SD rats were selected and divided into the normal control group( normal SD rats without any treatment),OA model group and TDP-43-m MSCs + OA group,20 rats in each group. The rat OA model was established by collagen enzyme injection. TDP-43-m MSCs + OA group rats were subjected to TDP-43-m MSCs transplantation intervention. Western blot was used to detect the protein expression of TDP-43,RACK1,MTK1,MAPKKK,formation of cytoplasmic SGs,and the protein changes of JNK and p38 MAPK signaling pathways. The secretion of TNF-α and IL-1β in serum of rats was determined by ELISA. Results Compared with the normal control group,the expression of TDP-43 protein in the cartilage tissue of rats in the OA model group was significantly decreased. The protein expression of TDP-43 in the cartilage tissue of rats in the TDP-43-m MSCs + OA group was significantly higher than that of the normal control group and the OA model group,which indicates that TDP-43-m MSCs transplantation was successful. The amount of cytoplasmic SGs in the cartilage tissue of rats in the OA model group was significantly decreased compared with the normal control group. The TDP-43-m MSCs + OA group was significantly higher than model group. The expression of RACK1,JNK and p38 MAPK,MTK1 and MAPKKK phosphorylation,the secretion of TNF-α and IL-1β in serum in the cartilage tissue of rats in the OA model group were significantly higher than normal control group.the TDP-43-m MSCs + OA group were significantly lower than model group. Conclusion Overexpression of TDP-43 gene can interfere with the secretion of inflammatory factors and inhibit the activation of JNK and p38 MAPK signaling pathways dependent on ischemic hypoxia stress.Thus,the molecular mechanism of chondrocytopathic lesions is reversed,which provided a new theoretical basis for the treatment of OA.更多还原