新风胶囊含药血清通过调控环状RNA Cbl原癌基因B(circ-CBLB)影响类风湿关节炎滑膜成纤维细胞增殖和凋亡  

Serum from Xinfeng Capsule-treated rats affect the proliferation and apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by regulating circular RNA Cbl proto-oncogene B(circ-CBLB)

作　　者：李舒 万磊[1] 刘健[1] 黄传兵[1] 李方泽 程静 胡赛赛 陈莹莹 朱子衡 LI Shu;WAN Lei;LIU Jian;HUANG Chuanbing;LI Fangze;CHENG Jing;HU Saisai;CHEN Yingying;ZHU Ziheng(Department of Rheumatology,First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学第一附属医院风湿科,安徽合肥230031

出　　处：《细胞与分子免疫学杂志》2024年第9期792-799,共8页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家中医药管理局第七批全国老中医药专家学术经验继承项目(国中医药人教函[2022]76号)。国家自然科学基金面上项目(81973655,82274501);安徽省高校优秀青年人才支持计划项目(皖教秘人[2022]11号);新安医学教育部重点实验室(2020xayx04);安徽省高校自然科学研究项目(KJ2020A0397);安徽省新时代育人质量工程项目(2022xscx103)。

摘　　要：目的探讨新风胶囊(XFC)含药血清通过调控环状RNA Cbl原癌基因B(circ-CBLB)对类风湿关节炎成纤维细胞样滑膜细胞(RA-FLS)增殖、凋亡的影响。方法XFC灌胃大鼠,制备含药血清。人RA-FLS来源于RA患者滑膜组织,用100μL的10 ng/mL肿瘤坏死因子α(TNF-α)刺激RA-FLS建立模型。构建pcDNA3.1-circ-CBLB及阴性对照,分别转染至RA-FLS中。实验分为对照组、TNF-α处理的RA-FLS组、XFC处理的RA-FLS组、pcDNA3.1-circ-CBLB-NC转染的RA-FLS组(过表达空转组)、pcDNA3.1-circ-CBLB转染的RA-FLS组(过表达组)、pcDNA3.1-circ-CBLB联合XFC处理的RA-FLS组(过表达给药组)。采用CCK-8法检测细胞活力,采用流式细胞术检测细胞周期与凋亡,采用实时定量PCR检测circ-CBLB表达水平,采用ELISA检测抗炎因子白细胞介素4(IL-4)、IL-10,促炎因子IL-6、TNF-α水平。结果XFC最佳含药血清量为200 mL/L,处理时间为72 h;与同一时间模型组相比,XFC组、过表达组、过表达给药组的细胞活力均下降。与模型组比较,XFC组、过表达组、过表达给药组circ-CBLB表达水平升高、凋亡率升高,S期和G2期细胞比例增加,IL-4、IL-10含量升高,IL-6、TNF-α含量降低。结论XFC处理上调RA-FLS中circ-CBLB表达,提高抗炎因子的水平,降低促炎因子的水平,抑制RA-FLS的细胞活力,提高其凋亡率,延长细胞周期,抑制RA-FLS增殖并促进其凋亡。Objective To explore the effect of serum from Xinfeng Capsule(XFC)-treated rats on the proliferation and apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis(RA-FLS)by regulating the circular RNA Cbl oncogene B(circ-CBLB).Methods XFC was administered orally to rats to prepare drug-containing serum.Human RA-FLS were stimulated with 100μL of 10 ng/mL tumor necrosis factor-alpha(TNF-α)to establish the model.pcDNA3.1-circ-CBLB and negative control were constructed and transfected into RA-FLS.The experiment was divided into six groups:control group,TNF-αtreated RA-FLS group,XFC treated RA-FLS group,pcDNA3.1-circ-CBLB-NC,pcDNA3.1-circ-CBLB(overexpression group),pcDNA3.1-cicr-CBLB combined with XFC treated group(overexpression+XFC group).Cell viability was assessed by CCK-8 assay;cell cycle and apoptosis by flow cytometry,and the expression levels of circ-CBLB in each group by real-time quantitative PCR.The levels of inflammatory cytokines interleukin 4(IL-4)、IL-10、IL-6 and TNF-αwere measured by ELISA.Results The optimal serum was 200 mL/L,and the treatment time was 72 hours;Compared with the model group at the same time point,the cell viability of XFC group,overexpression group,and overexpression+XFC group were lower,while the expression level and apoptosis rate of circ-CBLB were higher.The proportion of cells in S phase and G2 phase was higher.Additionally,the levels of IL-4 and IL-10 were higher,while the levels of IL-6 and TNF-αwere lower.Conclusion XFC treatment upregulates the expression of circ-CBLB in RA-FLS,increases anti-inflammatory cytokines,decreases pro-inflammatory cytokines,inhibits the viability of RA-FLS,increases apoptosis rate,extends the cell cycle,suppresses the proliferation of RA-FLS,and promotes its apoptosis.

关 键 词：新风胶囊(XFC) 环状RNA Cbl原癌基因B(circ-CBLB) 成纤维细胞样滑膜细胞(FLS) 细胞增殖 细胞凋亡 细胞因子 

分 类 号：R593.22[医药卫生—内科学] R285.5[医药卫生—临床医学] Q255[生物学—细胞生物学] R684.3R364.5