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目的研究类胰蛋白酶(tryptase)对MH7A类风湿性关节炎(RA)滑膜成纤维细胞表面蛋白酶激活受体2(PAR-2)、Rho信号通路及细胞凋亡的影响。方法采用流式细胞术检测MH7A细胞表面受体PAR-2的表达;异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测细胞的凋亡情况;采用Pull-down及Western blot法检测MH7A细胞Rho激酶的表达变化。结果类胰蛋白酶能够上调MH7A细胞表面PAR-2的表达,并剂量依赖性的抑制Fas介导的MH7A细胞的凋亡;同时,PAR-2抑制剂FSLLRY-NH2能够显著降低类胰蛋白酶对MH7A细胞的抗凋亡作用,其作用与活化Rho激酶的表达增加有关。结论类胰蛋白酶通过上调PAR-2和活化Rho激酶发挥很强的抗MH7A细胞凋亡的作用。
Objective To study the effects of tryptase on the expression of PAR-2, Rho signaling pathway and apoptosis in synovial fibroblasts of MH7A-type RA rheumatoid arthritis (RA). Methods Flow cytometry was used to detect the expression of PAR-2 on the surface of MH7A cells. Fluorescein-labeled annexinⅤ-FITC / PI was detected by flow cytometry The apoptosis of MH7A cells was detected by pull-down and Western blot. Results Tryptase could up-regulate the expression of PAR-2 on the surface of MH7A cells and dose-dependently inhibit Fas-mediated apoptosis of MH7A cells. Meanwhile, PAR-2 inhibitor FSLLRY-NH2 could significantly reduce the effect of tryptase on MH7A cells Anti-apoptotic effect, the role of activated Rho kinase expression increased. Conclusion Tryptase exerts a strong anti-apoptotic effect on MH7A cells by upregulating PAR-2 and activating Rho kinase.