耐受性树突状细胞可抑制胶原诱导性关节炎大鼠脾脏细胞的焦亡  

Tolerogenic dendritic cells can inhibit pyroptosis of spleen cells in collagen-induced arthritis rats

作　　者：龙调玉 包伦敏 万秀方 李红红 张云东[2] 蒋红梅 Long Tiaoyu;Bao Lunmin;Wan Xiufang;Li Honghong;Zhang Yundong;Jiang Hongmei(Department of Clinical Microbiology and Immunology,School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550025,Guizhou Province,China;Department of Laboratory Medicine,People’s Hospital of Anshun City,Anshun 561000,Guizhou Province,China)

机构地区：[1]贵州医科大学医学检验学院临床微生物学及免疫学教研室,贵州省贵阳市550025 [2]安顺市人民医院医学检验科,贵州省安顺市561000

出　　处：《中国组织工程研究》2023年第33期5363-5369,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区科学基金(81960393),项目负责人:蒋红梅;贵州省普通本科高校自然科学研究创新群体项目(黔教合KY字[2021]016),项目领衔人:蒋红梅。

摘　　要：背景:课题组前期利用核因子κB寡核苷酸诱骗剂成功构建了抗原特异性耐受性树突状细胞,并用其对胶原诱导性关节炎大鼠模型进行了有效干预,但治疗机制仍有待进一步研究。目的:探讨耐受性树突状细胞对胶原诱导性关节炎大鼠脾脏细胞焦亡的影响。方法:提取骨髓源树突状细胞,利用核因子κB寡核苷酸诱骗剂构建耐受性树突状细胞。取9只SD大鼠,采用随机数字表法随机分为正常组、模型组、治疗组,每组3只,模型组、治疗组建立胶原诱导性关节炎模型(初次免疫:左足跖部皮内注射牛Ⅱ型胶原乳剂;加强免疫:初次免疫后第14天,尾根部多点注射牛Ⅱ型胶原乳剂),初次免疫后第20天分别尾静脉注射生理盐水、耐受性树突状细胞,初次免疫后第35天,进行相关检测。结果与结论:(1)关节炎指数评分:治疗组大鼠关节炎指数评分低于模型组(P<0.05);(2)足踝关节大体与病理观察:模型组大鼠踝关节严重肿胀,治疗组大鼠踝关节轻度肿胀;苏木精-伊红染色显示,与正常组相比,模型组大鼠足踝关节滑膜明显增生,伴有炎性细胞浸润和骨侵蚀;与模型组相比,治疗组大鼠足踝关节滑膜增生减低、骨侵蚀和炎性细胞浸润减少;(3)血清白细胞介素1β水平:模型组大鼠血清白细胞介素1β水平高于正常组(P<0.05),治疗组大鼠血清白细胞介素1β水平低于模型组(P<0.05);(4)脾脏相关蛋白表达:Western blot检测显示,与正常组相比,模型组大鼠脾脏NLRP3、pro-caspase-1、caspase-1、GSDMD-NT、白细胞介素1β前体、白细胞介素1β的蛋白表达均增高(P<0.05);与模型组大鼠相比,治疗组大鼠脾脏NLRP3、pro-caspase-1、caspase-1、GSDMD-NT、白细胞介素1β前体、白细胞介素1β的蛋白表达均下降(P<0.05);(5)结果表明,耐受性树突状细胞可能通过抑制脾脏细胞焦亡和促炎细胞因子白细胞介素1β的释放,缓解胶原诱导性关节炎模型大鼠的炎症�BACKGROUND:In our previous study,we successfully constructed antigen-specifc tolerogenic dendritic cells using nuclear factor-kappa B oligodeoxynucleotide decoy,and used them to effectively intervene a collagen-induced arthritis rat model.However,the therapeutic mechanism remains to be further studied.OBJECTIVE:To investigate the effect of tolerogenic dendritic cells on pyroptosis of spleen cells in collagen-induced arthritis rats.METHODS:Bone marrow-derived dendritic cells were extracted and used to construct tolerogenic dendritic cells using nuclear factor-kappa B oligodeoxynucleotide decoy.Nine Sprague-Dawley rats were randomly divided into control group,model group,and treatment group,with three rats in each group.The latter two groups were used to establish rheumatoid arthritis models.Initial immunization was subcutaneous injection of bovine typeⅡcollagen emulsion on the le?t foot plantar;booster immunization was multiple injections of bovine typeⅡcollagen emulsion at the base of the tail on the 14thday a?ter the initial immunization.Rats in the model and treatment groups were given normal saline and tolerogenic dendritic cells into the tail vein,respectively,on the 20thday a?ter the initial immunization.Relevant measurements were performed on the 35thday a?ter the initial immunization.RESULTS AND CONCLUSION:(1)Compared with the model group,arthritis index score in the treatment group decreased signifcantly(P<0.05).(2)Gross and pathological observations showed the ankle joints of the rats were severely swollen in the model group and slightly swollen in the treatment group.Hematoxylin-eosin staining results showed that compared with the control group,the synovium of the ankle joint had obvious hyperplasia,accompanied by inflammatory cell infltration and bone erosion;compared with the model group,the treatment group had reduced synovial hyperplasia,bone erosion and inflammatory cell infltration of the ankle joint.(3)Serum interleukin 1βlevel was higher in the model group than the control group(P<0.01)and

关 键 词：耐受性树突状细胞 胶原诱导性关节炎 NLRP3 白细胞介素1Β 细胞焦亡 类风湿关节炎 

分 类 号：R452[医药卫生—治疗学] R363[医药卫生—临床医学] R593.2