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目的探讨糖代谢关键酶在类风湿关节炎(RA)中的表达,进一步探讨糖代谢与RA的相关性。方法取牛Ⅱ型胶原诱导大鼠(CIA组)和正常大鼠(NCR组)膝关节滑膜组织,提取总RNA,采用Rat Glucose Metabolism RT2ProfilerTMPCR Array筛选表达不一致基因;采用RT-PCR法对CIA组和NCR组进行mRNA水平的检测,采用RT-PCR法和Western blotting法检测RA组和骨关节炎组(OA组)滑膜组织中基因mRNA水平和蛋白水平的表达。收集RA血清(RAB组)和正常献血者血清(NCB组),采用ELISA法检测血清中的表达水平。结果 PCR Array检测显示,CIA组烯醇化酶(ENO1)、己糖激酶2(HK2)、磷酸甘油酸激酶-1(PGK1)较NCR组表达上调(P<0.05),磷酸烯醇式丙酮酸羧激酶-1(PCK1)和丙酮酸脱氢酶激酶-4(PDK4)表达下调(P<0.05);RT-PCR法和Western blotting法检测显示,RA组与OA组相比,ENO1和PGK1表达上调(P<0.05),ELISA法检测结果显示,RAB组与NCB组相比,PGK1表达升高(P<0.05)。结论牛Ⅱ型胶原诱导大鼠和RA滑膜组织中糖代谢关键酶ENO1和PGK1表达增高,RA血液中PGK1表达增加,均提示RA中糖代谢活动增强,导致相关基因的表达异常。
Objective To investigate the expression of key enzymes of carbohydrate metabolism in rheumatoid arthritis (RA) and to explore the relationship between glucose metabolism and RA. Methods The total RNA was extracted from the synovial tissue of rats induced by collagen type Ⅱ collagen (CIA) and normal rats (NCR group), and the non-identical genes were screened by using Rat Glucose Metabolism RT2 Profiler TMPCR Array. And NCR group. The mRNA and protein levels of synovial tissue in RA group and OA group were detected by RT-PCR and Western blotting. RA serum (RAB group) and normal blood donor serum (NCB group) were collected, and the expression level in serum was detected by ELISA. Results PCR Array showed that ENO1, HK2 and PGK1 were up-regulated in CIA group (P <0.05), and phosphoenolpyruvate carboxylase The expression of PCK1 and PDK4 were down-regulated (P <0.05). Compared with OA group, the expression of ENO1 and PGK1 were up-regulated in RA group compared with OA group (P <0.05). The results of ELISA showed that the expression of PGK1 in RAB group was higher than that in NCB group (P <0.05). CONCLUSION: The expression of ENO1 and PGK1, the key enzymes of glucose metabolism in rat and RA synovium, is increased and the expression of PGK1 in RA blood is increased, which indicates that the activity of glucose metabolism in RA is increased, resulting in the abnormal expression of related genes.