導讀
Rheumatology & Autoimmunity
山西醫科大學第二醫院李小峰教授團隊在Rheumatology & Autoimmunity 線上發表最新研究，深入探讨了雷帕黴素對類風濕關節炎（RA）患者滑膜細胞（FLSs）的增殖抑制和凋亡誘導作用，為RA的臨床治療提供了新的分子機制和治療政策。
研 究 背 景
類風濕關節炎（RA）是一種慢性發炎性自身免疫性疾病，其特征是關節滑膜的過度增生和發炎，導緻關節損傷和功能障礙。RA的病理過程中，滑膜細胞（FLSs）的異常增殖和抗凋亡特性起着關鍵作用。李小峰教授團隊采用細胞計數試劑盒-8檢測雷帕黴素對RA-FLSs增殖的影響，并通過實時聚合酶鍊反應和蛋白質印迹分析，探讨了雷帕黴素通過AKT/mTORC1信号通路調控RA-FLSs的分子機制。
研 究 結 果
研究發現，雷帕黴素能夠顯著抑制RA-FLSs的增殖，并以劑量依賴性方式誘導細胞凋亡。在分子水準上，雷帕黴素顯著上調了促凋亡蛋白Bax和caspase家族成員的表達，同時降低了抗凋亡蛋白Bcl-2的表達。此外，雷帕黴素還抑制了AKT/mTORC1信号通路的關鍵分子，包括p-AKT、p-mTOR、p-S6K和p-4EBP1的磷酸化水準，進而抑制了RA-FLSs的增殖并促進了細胞凋亡。
Figure 1Identification of RA-FLSs. Flow cytometry to check the expression of cell surface markers CD68, CD90, FAP-α on RA-FLSs. (A) Blank. (B) RA-FLSs. APC, allophycocyanin; FAP, fibroblast activation protein; PE, phycoerythrin; RA-FLS, rheumatoid arthritis fibroblast-like synoviocytes.
Figure 2 Cell inhibition rate of rheumatoid arthritis fibroblast-like synoviocytes after intervention with different concentrations of rapamycin. (A) Cell inhibition rate after 24 h intervention. (B) Cell inhibition rate after 48 h intervention. **p < 0.01, rapamycin group (1, 5, 10, 15, 20, 25 nmol/L) versus the control group.
Figure 3 Cell apoptosis determined by flow cytometry. (A) Flow rate of apoptosis in control group. (B) Flow rate of apoptosis in rapamycin group (10 nmol/L). (C) Histogram of apoptosis rate of RA-FLSs. ***p < 0.001. ADC, apparent diffusion coefficient; FITC, fluorescein isothiocyanate; FL1, plotting green fluorescence; FL4, red fluorescence intensity; PI, propidium iodide; RA-FLS, rheumatoid arthritis fibroblast-like synoviocytes.
Figure 4 RA-FLSs were treated with rapamycin (10 nmol/L) for 48 h. (A) Relative expression of Bax and Bcl-2 mRNA. (B) Western blotting analysis was performed to detect the protein levels of Bax and Bcl-2. (C) Rapamycin increases Bax, caspase 3, and caspase 9 expression levels and decreases Bcl-2 expression level in RA-FLSs. *p < 0.05; **p < 0.01. Bax, Bcl-2-associated X protein; Bcl-2, B cell lymphoma 2; RA--FLS, rheumatoid arthritis-fibroblast-like synoviocytes.
Figure 5 RA-FLSs were treated with rapamycin (10 nmol/L) for 48 h. (A) Expression level of CD1, CDK2, and β-actin in RA-FLSs by Western blotting analysis. (B) Quantitative analysis of CDK2, CD1 expression in RA-FLSs by Western blotting analysis. *p < 0.05, **p < 0.01. CDK2, cyclin-dependent kinase 2; RA-FLS, rheumatoid arthritis fibroblast-like synoviocytes.
Figure 6 RA-FLSs were treated with rapamycin (10nmol/L) for 48 h. (A) Expression level of AKT and mTOR mRNA. (B) Relative expression of p-AKT/AKT, p-mTOR/mTOR, p-S6K/S6K, p-4EBP1/4EBP1; (C) Quantitative analysis of AKT/mTORC1 pathway expression in RA-FLSs by Western blotting analysis, **p < 0.01, *p < 0.05. 4EBP1, 4E-binding protein 1; AKT, protein kinase B; Bax, Bcl-2-associated X protein; Bcl-2, B cell lymphoma 2; mTOR, mammalian target of rapamycin; RA-FLSs, rheumatoid arthritis-fibroblast-like synoviocytes; S6K, S6 kinase.
結 論
該研究系統性地揭示了雷帕黴素通過靶向AKT/mTORC1信号通路，對RA-FLSs的增殖和凋亡具有顯著的調控作用。這一發現為RA的臨床治療提供了新的分子靶點和治療政策。李小峰教授指出，雷帕黴素作為一種新型的免疫調節劑，不僅能通過調控Treg細胞的增殖和穩态誘導免疫平衡，亦可以直接作用于RA的滑膜細胞發揮治療作用，其在RA等諸多自身免疫性疾病的治療中的潛力值得關注。
歡 迎 引 用
How to cite: Zhang S, Hu X, Su Q, et al. Rapamycin suppresses rheumatoid arthritis fibroblast synovial cell proliferation and induces apoptosis via the AKT/mTORC1 pathway. Rheumatol & Autoimmun. 2024; published ahead of print. doi:10.1002/rai2.12120.
作者簡介
李小峰
山西省風濕免疫科首席學科帶頭人二級教授，主任醫師，博士生導師享受國務院政府特殊津貼山西省五一勞動獎章獲得者擁有“國之名醫•卓越建樹”及“山西名醫”、“山醫名師”稱号原“136”興醫工程領軍臨床專科負責人《免疫微生态學》專著主編曾任中華醫學會風濕病學分會 常委現任中國醫師協會風濕免疫科醫師分會 副會長中華醫學會山西分會 常務理事山西省醫師協會 常委風濕免疫科醫師分會 會長
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