来源：风湿病与关节炎,2023,12（4）:1-4,9.

电针调控lncRNA NEAT1减轻类风湿关节炎滑膜成纤维细胞炎症反应的研究

（基础研究）

黄志强¹，涂海水^2,3，李莹莹³，郑志煌³，李佳璇³，程镇达³，苏稼夫¹

　　【摘　要】目的：观察电针对调控lncRNA NEAT1减轻类风湿关节炎大鼠滑膜成纤维细胞炎症反应的影响。方法：将16只2月龄大鼠应用随机数字表法分为空白对照组和电针组，每组8只。空白对照组大鼠未接受干预，电针组大鼠予以电针干预，分别制备空白对照组、电针组血清。予以连续酶消化法获取3周龄SD大鼠滑膜成纤维细胞，波形蛋白免疫组化染色进行鉴定；再将6孔板中已培养的滑膜成纤维细胞随机分为空白对照组、模型对照组、电针组成纤维细胞。空白对照组成纤维细胞予以空白对照组血清干预，模型对照组成纤维细胞予以含10 ng·mL^-1 白细胞介素-1β（IL-1β）的空白对照组血清诱导滑膜成纤维细胞致炎，电针组成纤维细胞予以含10 ng·mL^-1 IL-1β的电针组血清干预。Real-time PCR检测各组滑膜成纤维细胞中lncRNA NEAT1水平变化；Western blot检测各组滑膜成纤维细胞基质金属蛋白酶-3（MMP-3）、V-Rel网状内皮增生病毒癌基因同源物A（RELA）、肿瘤坏死因子-α（TNF-α）蛋白表达含量。结果：经波形蛋白免疫组化染色后，大鼠滑膜成纤维细胞波形蛋白染色为阳性，证实成纤维细胞提取成功。与空白对照组比较，模型对照组中lncRNA NEAT1的相对表达水平显著升高（P < 0.05）；与模型对照组比较，电针组中lncRNA NEAT1相对表达水平显著降低（P < 0.05）。Western blot结果显示，与模型对照组比较，电针组MMP-3、RELA、TNF-α蛋白表达含量显著降低（P < 0.05）。结论：电针可通过调控lncRNA NEAT1减轻类风湿关节炎大鼠滑膜成纤维细胞炎症反应。

　　【关键词】　类风湿关节炎；滑膜成纤维细胞；炎症反应；电针；大鼠

Study on Electroacupuncture Regulating lnc NEAT1 to Alleviate the Inflammatory Response of Rheumatoid Arthritis Synovial Fibroblasts

HUANG Zhi-qiang,TU Hai-shui,LI Ying-ying,ZHENG Zhi-huang,LI Jia-xuan,CHENG Zhen-da,SU Jia-fu

　　【ABSTRACT】Objective:To observe the effect of electroacupuncture on reducing the inflammatory response of synovial fibroblasts in rheumatoid arthritis rats by regulating lncRNA NEAT1.Methods:Sixteen 2-month-old rats were randomly divided into a blank control group and an electroacupuncture group using a random number table method,with 8 rats in each group.Rats in the blank control group were not given intervention,while those in the electroacupuncture group were given electroacupuncture intervention,and serum from the two groups was prepared separately.The synovial fibroblasts of 3-week-old SD rats were obtained by continuous enzymatic digestion,and identified by vimentin immunohistochemical staining.Then,the cultured synovial fibroblasts in the 6-well plate were randomly divided into a blank control group,a model control group,and an electroacupuncture group.The blank control group was intervened with serum from the blank control group,while the model control group was induced inflammation by serum with 10 ng·mL^-1 interleukin-1β（IL-1β）from the blank control group,and the electroacupuncture was intervened by serum containing 10 ng·mL^-1 IL-1β from the electroacupuncture group.Real-time PCR was used to detect the level changes of lncRNA NEAT1 in synovial fibroblasts of each group.Western blot was used to detect protein expression contents of MMP-3,RELA and TNF-α.Results:After vimentin immunohistochemical staining,the rat synovial fibroblasts were positive for vimentin staining,which confirmed the successful extraction of fibroblasts.Compared with the blank control group,the relative expression level of lncRNA NEAT1 in the model control group was significantly increased（P < 0.05）.Compared with the model control group,the relative expression level of lncRNA NEAT1 in the electroacupuncture group was significantly reduced（P < 0.05）.Western blot results showed that compared with the model control group,the protein expression contents of MMP-3,RELA,and TNF-α in the electroacupuncture group significantly decreased（P < 0.05）.Conclusion:Electroacupuncture can alleviate the inflammatory response of synovial fibroblasts in rheumatoid arthritis rats by regulating lncRNA NEAT1.

　　【Keywords】 rheumatoid arthritis;synovial fibroblasts;inflammatory response;electroacupuncture;rats