Page 61 - 《中国药房》2023年15期

β-谷甾醇对类风湿性关节炎滑膜成纤维细胞功能的影响及机制
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谷慧敏 ，孟庆良 ，左瑞庭 ，展俊平 ，马俊福 ，刘亚伟（1.河南省中医院风湿病科，郑州 450002；2.河南中医
药大学骨伤学院，郑州 450002）
中图分类号 R965 文献标志码 A 文章编号 1001-0408（2023）15-1847-06
DOI 10.6039/j.issn.1001-0408.2023.15.11
摘 要 目的 研究β-谷甾醇对类风湿性关节炎（RA）滑膜成纤维细胞MH7A功能的影响及其机制。方法 使用网络药理学筛选
β-谷甾醇的作用靶点及治疗 RA 的靶点，两者取交集后进行拓扑分析寻找治疗 RA 的最关键靶点。用不同浓度（0、5、10、20、40
μmol/L）的 β-谷甾醇处理 MH7A 细胞并用 CCK-8 法检测细胞活性，筛选 β-谷甾醇的最适给药浓度。采用 10 ng/mL 肿瘤坏死
因子（TNF-α）体外诱导MH7A细胞后，加入β-谷甾醇（最适给药浓度）处理。CCK-8法和EdU法检测细胞增殖能力；划痕实验和
Transwell侵袭法分别检测细胞迁移及侵袭能力；酶联免疫吸附测定（ELISA）法检测细胞上清液中白细胞介素1β（IL-1β）和IL-6的
水平；qRT-PCR法和Western blot法分别检测过氧化物酶体增殖物激活受体α（PPARα）mRNA和蛋白表达水平。MH7A细胞转染
PPARα小干扰RNA，并通过上述实验方法检测PPARα敲低后β-谷甾醇对MH7A细胞增殖、迁移、侵袭、炎症因子分泌及PPARα蛋
白表达的影响。结果 PPARα是β-谷甾醇治疗RA的最关键靶点，β-谷甾醇的最适给药浓度为20 μmol/L。与模型组相比，β-谷甾
醇组MH7A细胞活力显著降低、细胞增殖数显著减少（P＜0.05），细胞迁移率显著降低、细胞侵袭数显著减少（P＜0.05），IL-1β水
平、IL-6水平均显著降低（P＜0.05），PPARα mRNA和蛋白表达水平均显著增加（P＜0.05）。与阴性对照小干扰RNA组相比，敲低
PPARα后细胞活力上升35.6%（P＜0.05），细胞增殖数、细胞迁移率和细胞侵袭数均显著增加（P＜0.05），IL-1β水平及IL-6水平均
显著升高（P＜0.05）。结论 β-谷甾醇可以抑制TNF-α诱导的MH7A细胞的增殖、迁移、侵袭和炎症因子分泌，其作用机制与激活
PPARα通路有关。
关键词 β-谷甾醇；类风湿性关节炎；过氧化物酶体增殖物激活受体α；增殖；迁移；侵袭；炎症因子

Effects of β-sitosterol on the function of synovial fibroblasts in rheumatoid arthritis and its mechanism
GU Huimin ，MENG Qingliang ，ZUO Ruiting ，ZHAN Junping ，MA Junfu ，LIU Yawei （1. Dept. of
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Rheumatology， Henan Province Hospital of TCM， Zhengzhou 450002， China；2. College of Orthopedics and
Traumatology， Henan University of Chinese Medicine， Zhengzhou 450002， China）
ABSTRACT OBJECTIVE To investigate the effects of β -sitosterol on the function of rheumatoid arthritis （RA） fibroblastic
synoviocytes MH7A cells and its mechanism. METHODS Network pharmacology was adopted to screen the targets of β-sitosterol
and the targets for the treatment of RA. After the intersection of them， topological analysis was performed to find the most critical
target in the treatment of RA. MH7A cells were treated with different concentrations （0， 5， 10， 20， 40 μmol/L） of β-sitosterol， and
CCK-8 was used to assay cell viability for screening the optimal concentration of β-sitosterol. MH7A cells were induced by 10 ng/mL
TNF-α in vitro and treated with β-sitosterol （the optimum concentration）. CCK-8 and EdU were used to detect the ability of cell
proliferation. Scratch experiment and Transwell invasion assay were used to analyze cell migration and invasion. The levels of
interleukin-1β （IL-1β） and IL-6 in cell supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA
and protein expressions of peroxisome proliferator-activated receptor α （PPARα） were measured by qRT-PCR and Western blot，
respectively. The siRNA targeting PPARα was transfected into MH7A cells， and the effects of β-sitosterol on cell proliferation，
migration， invasion， the secretion of inflammatory factors and the expression of PPARα after PPARα knockdown were detected by
the above experimental methods. RESULTS PPARα was the most critical target of β-sitosterol in the treatment of RA. The optimal
concentration of β-sitosterol was 20 μmol/L. Compared with model group， β-sitosterol decreased the viability of MH7A cells， and
the number of proliferating cells also decreased significantly （P＜0.05）； the cell migration rate and the number of cell invasion
decreased significantly （P＜0.05）. The levels of IL-1β and IL-
Δ 基金项目 河南省卫生健康委2021年中医药传承与创新人才工 6 were also significantly decreased （P＜0.05）， and the mRNA
程（仲景工程）中医药拔尖人才资助项目（No. 豫卫中医函〔2021〕15 and protein expression levels of PPARα were significantly
号）；河南省中医药科学研究专项课题（No.2019ZY2060） increased （P＜0.05）. Compared with negative control small
＊第一作者 副主任医师，硕士。研究方向：中医药防治风湿类疾
interfering RNA group， after PPARα knockdown， the cell
病。电话：0371-53312126。E-mail：gensheng200@163.com
# 通信作者 主任医师。研究方向：中医药防治风湿类疾病。电 viability increased by about 35.6% （P＜0.05）， the number of
话：0371-53312126。E-mail：mql678@163.com cell proliferation， the cell migration rate and the number of

中国药房 2023年第34卷第15期 China Pharmacy 2023 Vol. 34 No. 15 · 1847 ·