Liu Yang, Liu Ping-Ping, Liu Lei, Zheng Xiao-Shuo, Zheng Hui, Yang Cheng-Cheng, Luobu Ci-Ren, Liu Ye
Department of Ophthalmology, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, Guangdong Province, China.
Department of Ophthalmology, the First Hospital of Jilin University, Changchun 130021, Jilin Province, China.
Int J Ophthalmol. 2018 Jul 18;11(7):1108-1113. doi: 10.18240/ijo.2018.07.06. eCollection 2018.
To determine if triptolide influences the contractility and fibronectin production in human Tenon fibroblasts (HTFs).
HTFs were cultured in type I collagen gels with or without transforming growth factor beta (TGF-β) and/or triptolide. The diameter of the collagen gel was used to measure contraction. Immunoblot analysis was used to quantify myosin light chain (MLC) phosphorylation and integrin expression. Laser confocal fluorescence microscopy was used to monitor the formation of actin stress fibers. Fibronectin production was measured with an enzyme immunoassay.
Triptolide inhibition of contraction in TGF-β-induced collagen gel mediated by HTFs was dose-dependent and statistically significant at 3 nmol/L (<0.05) and maximal at 30 nmol/L and significantly time dependent at 2d (<0.05). Triptolide reduced TGF-β-induced expression of integrins α5 and β1, phosphorylation of MLC, and formation of stress fibers in HTFs. Furthermore, the inhibition of triptolide on the attenuated TGF-β-induced production of fibronectin by HTFs was concentration-dependent and significant at 1 nmol/L (<0.05) and maximal at 30 nmol/L.
Triptolide suppress the contractility of HTFs induced by TGF-β and the production of fibronectin by these cells. It is promising that triptolide treatment may possibly inhibit scar formation after glaucoma filtration surgery.
确定雷公藤甲素是否影响人Tenon囊成纤维细胞(HTFs)的收缩性和纤连蛋白生成。
将HTFs培养于含有或不含有转化生长因子β(TGF-β)和/或雷公藤甲素的I型胶原凝胶中。用胶原凝胶的直径来测量收缩。免疫印迹分析用于定量肌球蛋白轻链(MLC)磷酸化和整合素表达。激光共聚焦荧光显微镜用于监测肌动蛋白应力纤维的形成。用酶免疫测定法测量纤连蛋白生成。
雷公藤甲素对HTFs介导的TGF-β诱导的胶原凝胶收缩的抑制呈剂量依赖性,在3 nmol/L时具有统计学意义(<0.05),在30 nmol/L时达到最大,且在2天时具有显著的时间依赖性(<0.05)。雷公藤甲素降低了TGF-β诱导的HTFs中整合素α5和β1的表达、MLC的磷酸化以及应力纤维的形成。此外,雷公藤甲素对HTFs减弱的TGF-β诱导的纤连蛋白生成的抑制呈浓度依赖性,在1 nmol/L时具有显著意义(<0.05),在30 nmol/L时达到最大。
雷公藤甲素抑制TGF-β诱导的HTFs的收缩性以及这些细胞的纤连蛋白生成。雷公藤甲素治疗可能抑制青光眼滤过术后瘢痕形成,这是很有前景的。