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.
Department of Rheumatology, First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2024 Sep;40(9):792-799.
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 100 μL 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-α treated 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-α were 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-α were 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.
目的 通过调控环状RNA Cbl原癌基因B（circ-CBLB），探讨新风胶囊（XFC）含药血清对类风湿关节炎成纤维样滑膜细胞（RA-FLS）增殖和凋亡的影响。方法 对大鼠灌胃给予XFC制备含药血清。用100 μL 10 ng/mL肿瘤坏死因子-α（TNF-α）刺激人RA-FLS建立模型。构建pcDNA3.1-circ-CBLB和阴性对照并转染至RA-FLS。实验分为六组：对照组、TNF-α处理的RA-FLS组、XFC处理的RA-FLS组、pcDNA3.1-circ-CBLB-NC、pcDNA3.1-circ-CBLB（过表达组）、pcDNA3.1-cicr-CBLB联合XFC处理组（过表达+XFC组）。采用CCK-8法检测细胞活力；通过流式细胞术检测细胞周期和凋亡情况，采用实时定量PCR检测各组circ-CBLB的表达水平。采用酶联免疫吸附测定法检测炎性细胞因子白细胞介素4（IL-4）、IL-10、IL-6和TNF-α的水平。结果 最佳血清浓度为200 mL/L，处理时间为72小时；与同一时间点的模型组相比，XFC组、过表达组和过表达+XFC组的细胞活力较低，而circ-CBLB的表达水平和凋亡率较高。S期和G2期细胞比例较高。此外，IL-4和IL-10水平较高，而IL-6和TNF-α水平较低。结论 XFC处理可上调RA-FLS中circ-CBLB的表达，增加抗炎细胞因子，降低促炎细胞因子，抑制RA-FLS的活力，增加凋亡率，延长细胞周期，抑制RA-FLS的增殖，并促进其凋亡。
