LncRNA H19对类风湿性关节炎的影响及作用机制  

Effects and mechanism of LncRNA H19 on arthritis model

作　　者：林晓 丁晨 朱伟[1] 朱晓冬[1] LIN Xiao(Department of Immunology,Mudanjiang Medical University,Mudanjiang 157011,China)

机构地区：[1]牡丹江医学院免疫教研室,黑龙江牡丹江157011

出　　处：《牡丹江医学院学报》2022年第1期7-10,共4页Journal of Mudanjiang Medical University

基　　金：黑龙江省自然科学基金重点项目(ZD2020H005)。

摘　　要：目的探讨长链非编码RNA H19(Long non-coding RNA H19,LncRNA H19)对类风湿性关节炎(rheumatoid arthritis,RA)的影响及作用机制。方法将24只6~8周龄雄性C57BL/6小鼠分为四组:Normal组、AIA组、AIA+AAV8-CMA组、AIA+AAV8-CMV-H19组,每组6只。Normal组在左侧踝关节周围皮下注射30μL生理盐水;其余组在相同位置注射30μL含结核分枝杆菌的弗氏完全佐剂(CFA);AIA+AAV8-CMA组于建模前3周,向AIA小鼠左侧踝关节注射5μL腺相关病毒(AAV8)空载;AIA+AAV8-CMV-H19组于建模前3周,向AIA小鼠左侧踝关节注射5μL携带H19的AAV8过表达LncRNA H19;肿胀测量仪记录各组关节肿胀程度;建模8 d后,处死小鼠收集踝关节标本;HE染色观察各组踝关节炎症病变,qRT-PCR检测各组TNF-α、CCL-2、CCR-7的表达情况,Western blot检测IRF5、p-IRF5表达。结果AIA+AAV8-CMV-H19组小鼠踝关节肿胀程度明显超过其他组(P<0.01);HE染色显示,AIA组、AIA+AAV8-CMA组小鼠关节滑膜明显增生,炎性细胞浸润,水肿明显,而AIA+AAV8-CMV-H19组可见大量炎性细胞浸润,水肿十分显著;与AIA+AAV8-CMA组相比,AIA+AAV8-CMV-H19组上调特征性炎症因子TNF-α、CCL-2、CCR-7的表达(P<0.01);Western Blot结果显示AIA+AAV8-CMV-H19组p-IRF5表达量较其他组明显升高(P<0.01)。结论过表达LncRNA H19加重RA病情,这一作用可能与转录因子IRF5有关。Objective To investigate the effects and mechanism of long-chain non-coding RNA H19(Long Non-Coding RNA H19(LncRNA H19)on arthritis model.Methods 24 C57BL/6 mice(6~8 week old and male)were divided into four groups:normal group,AIA group,AIA+AAV8-CMA group,and AIA+AAV8-CMV-H19 group,6 mice in each group.The normal group received 30μL normal saline surrounding left ankle joint,and other groups received 30μL Frego complete adjuvant(CFA)containing mycobacterium tuberculosis to establish adjuvant arthritis.During the first 3 weeks,the AIA+AAV8-CMA group was given ankle injection with 5μL gland-associated virus(AAV8)empty vector,and the AIA+AAV8-CMV-H19 group was given with 5μL gland-associated virus vector carrying H19 to overexpress LncRNA H19.8 days after modeling,the ankle synonymous specimens were collected,and the swelling measuring instrument was used recorded joint swelling of each group,the HE staining was used to observe ankle inflammatory lesions,qRT-PCR was used to detect TNF-α,CCL-2,CCR-7 expression,Western blot was used to detect the protein expression of IRF5 and P-IRF5.Results The results of the foot swelling in LncRNA H19 overexpression group was significantly obvious than that in the control group;HE stain results showed that the expression of LncRNA H19 promoted inflammatory cell infiltration and aggravated inflammatory disease compared to normal groups;Fluorescence quantitative PCR results showed that LncRNA H19 overexpress upregulated characteristic inflammatory factors TNF-α,CCL-2,CCR-7 compared to the AIA+AAV8-CMA group(P<0.01);Western blot results showed that phosphorylation level of transcription factor IRF5 in AIA+AAV8-CMV-H19 group was overexpressed(P<0.01).Conclusion The study found that the expression of LncRNA H19 aggravates RA condition,which may be related to the transcription factor IRF5.

关 键 词：RA LncRNA H19 IRF5 炎症因子 

分 类 号：R593.22[医药卫生—内科学]