山姜素对人类风湿关节炎成纤维细胞样滑膜细胞增殖、凋亡及炎症反应的影响  

The effect of alpinetin on the proliferation,apoptosis and inflammation of human rheumatoid arthritis fibroblast-like synoviocytes

作　　者：苗喜云[1] 王慧莲[1] 孟庆良[1] 左瑞庭[1] 马俊福[1] 范围 展俊平[1] MIAO Xi-Yun;WANG Hui-Lian;MENG Qing-Liang(Department of Rheumatism,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450000,Henan,China)

机构地区：[1]河南省中医院风湿病科,河南郑州450000

出　　处：《中国老年学杂志》2024年第18期4475-4480,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金(81804050);河南省中医药科学研究专项课题(2019ZY2060);河南省中医药传承与创新人才工程(仲景工程)中医药学科拔尖人才项目(2021年)。

摘　　要：目的探讨山姜素对人类风湿关节炎成纤维细胞样滑膜细胞增殖、凋亡及炎症应答的影响及潜在分子机制。方法采用肿瘤坏死因子(TNF)-α(10 ng/ml)刺激成纤维细胞样滑膜细胞系MH7A细胞24 h,建立体外炎症模型。将细胞分为对照组(无处理)、TNF-α组(加入10 ng/ml的TNF-α)和TNF-α+山姜素低、中、高剂量组(加入10 ng/ml的TNF-α培养12 h,再加入5、10或20μmol/L的山姜素培养24 h)。采用酶联免疫吸附试验(ELISA)和RT-q聚合酶链反应(PCR)分别检测细胞培养液上清液和细胞中白介素(IL)-6和IL-1β水平,噻唑蓝法(MTT)检测细胞增殖能力,TUNEL染色法和流式细胞术检测细胞凋亡,Western印迹检测B细胞淋巴瘤(Bcl)-2相关X蛋白(Bax)、Bcl-2、p-p65、磷酸化核因子(NF)-κB抑制蛋白(p-IκBα)、p65、IκBα和Toll样受体(TLR)4蛋白表达。结果与对照组比较,TNF-α组MH7A培养液上清液中IL-6和IL-1β含量及细胞中IL-6和IL-1βmRNA水平、细胞增殖率、p-p65/p65、p-IκBα/IκBα和TLR4蛋白表达明显升高(P<0.05);采用山姜素干预后,IL-6和IL-1βmRNA及含量、Bcl-2、p-p65/p65、p-IκBα/IκBα及TLR4蛋白表达、细胞增殖率明显降低,细胞凋亡率、Bax蛋白表达明显升高(P<0.05)。结论山姜素可降低TNF-α诱导的MH7A细胞增殖能力,并促进其凋亡,同时抑制炎症应答反应,其作用机制可能与抑制TLR4/NF-κB通路活化有关。Objective To explore the influence and potential molecular mechanisms of alpinetin on cell proliferation,apoptosis and inflammatory response in human rheumatoid arthritis fibroblast-like synoviocytes.Methods Fibroblast-like synovial cell line MH7A cells were stimulated with TNF-α(10 ng/ml)for 24 h to establish inflammatory cell model in vitro.Cells were divided into control group(no treatment),TNF-αgroup(treatment with 10 ng/ml TNF-α),TNF-α+alpinetin low,medium and high dose groups(treatment with 10 ng/ml TNF-αfor 12 h,and then treatment with 5,10 or 20μmol/L alpinetin for 24 h).Enzyme-linked immunosorbent assay(ELISA)and RT-q polymerase chain reaction(PCR)were used to detect the levels of interleukin(IL)-6 and IL-1βin the culture medium supernatant and cells,respectively.Cell proliferation was evaluated by thiazolyl blue(MTT)assay.TUNEL staining assay and flow cytometry were used to detect cell apoptosis.Western blot was used to estimate the protein expressions of B-cell lymphoma(Bcl)-2-associated X protein(Bax),Bcl-2,p-p65,p-nuclear factor(NF)-κB inhibits protein(IκBα),p65,IκBαand Toll-like receptor(TLR)4.Results Compared with the control group,IL-6,IL-1βcontents and IL-6,IL-1βmRNA levels in the culture medium superantant,cell proliferation rate,and protein expressions of p-p65/p65,p-IκBα/IκBαand TLR4 were significantly increased(P<0.05).After alpinetin treatment,IL-6,IL-1βmRNA and contents,the protein expressions of Bcl-2,p-p65/p65,p-IκBα/IκBαand TLR4 and cell proliferation rate were significantly decreased,cell apoptosis rate and Bax protein expression were significantly increased(P<0.05).Conclusions Alpinetin could decrease TNF-α-induced MH7A cell proliferation,facilitate cell apoptosis,and refrain inflammatory response,the mechanisms might be related to the inhibition on the activation of TLR4/NF-κB pathway.

关 键 词：山姜素 类风湿关节炎 成纤维样滑膜细胞 MH7A细胞 Toll样受体(TLR)4/NF-κB通路 

分 类 号：R285.5[医药卫生—中药学]