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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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| 篇名： | 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 |
| TITLE： | Effects of β-sitosterol on the function of synovial fibroblasts in rheumatoid arthritis and its mechanism |
| 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、40μmol/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10ng/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Westernblot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20μmol/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），PPARαmRNA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。 |
| 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-6 were also significantly decreased （P＜0.05）， and the mRNA 15 and protein expression levels of PPARα were significantly increased （P＜0.05）. Compared with negative control small interfering RNA group， after PPARα knockdown， the cell viability increased by about 35.6% （P＜0.05）， the number of cell proliferation， the cell migration rate and the number of cell invasion increased significantly （P＜0.05）， and the levels of IL-1β and IL-6 also increased significantly （P＜0.05）. CONCLUSIONS β-sitosterol could effectively inhibit the proliferation， migration， invasion and secretion of inflammatory factors in MH7A cells， the mechanism of which may be associated with activating PPARα pathway. |
| 期刊： | 2023年第34卷第15期 |
| 作者： | 谷慧敏；孟庆良；左瑞庭；展俊平；马俊福；刘亚伟 |
| AUTHORS： | GU Huimin，MENG Qingliang，ZUO Ruiting，ZHAN Junping，MA Junfu，LIU Yawei |
| 关键字： | 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 |
| KEYWORDS： | β-sitosterol； rheumatoid arthritis； peroxisome proliferator-activated receptor α； proliferation； migration； invasion； |
| 阅读数： | 67 次 |
| 本月下载数： | 2 次 |
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