Ye Xiaofeng, Ren Hui, Jiang Tingting, Zhang Ting, Li Gang
Eye Institute, EENT Hospital, Fudan University, Shanghai, China.
Laboratory of Center, EENT Hospital, Fudan University, Shanghai, China.
Histol Histopathol. 2022 Aug;37(8):757-767. doi: 10.14670/HH-18-427. Epub 2022 Feb 3.
This research was aimed to investigate whether diabetic blood-stasis syndrome had a relationship with ERK1/2-VEGF signaling pathway in rat retina Müller cells and Traditional Chinese Drugs designed for promoting blood circulation to remove blood stasis had effectiveness for diabetic retinopathy (DR) treatment.
Immunofluorescence was applied to determine purity of Müller cells. Müller cells were stimulated by blood serum obtained from rats with blood-stasis syndrome and then treated by Xuefu Zhuyu decoction. Western blot analysis, RT-PCR and ELISA were used to measure the expression of VEGF. Western blot analysis was used to determine the phosphorylation of ERK1/2. The status of AP-1 DNA binding activity was monitored by electrophoretic mobility shift assay (EMSA).
Stimulation of Müller cells by blood serum of rat with diabetic blood stasis increased the secretion of VEGF, activated ERK1/2 and AP-1 DNA-binding activity. And treatment of Xuefu Zhuyu decoction could weaken this phenomenon. What's more, ERK1/2 signaling pathway inhibitor U0126 also could inhibit the expression of VEGF.
Diabetic blood-stasis syndrome in theory of traditional Chinese Medicine has positive role in regulating ERK1/2-VEGF signaling pathway. Traditional Chinese drugs for promoting blood circulation to remove blood stasis would be an effective therapy to treat DR.
本研究旨在探讨糖尿病血瘀证与大鼠视网膜Müller细胞中ERK1/2-VEGF信号通路是否存在关联,以及活血化瘀中药对糖尿病视网膜病变(DR)治疗是否有效。
应用免疫荧光法测定Müller细胞纯度。用血瘀证大鼠血清刺激Müller细胞,然后用血府逐瘀汤进行处理。采用蛋白质免疫印迹法(Western blot分析)、逆转录聚合酶链反应(RT-PCR)和酶联免疫吸附测定(ELISA)检测血管内皮生长因子(VEGF)的表达。用蛋白质免疫印迹法分析检测细胞外信号调节激酶1/2(ERK1/2)的磷酸化水平。通过电泳迁移率变动分析(EMSA)监测激活蛋白-1(AP-1)DNA结合活性的状态。
糖尿病血瘀大鼠血清刺激Müller细胞可增加VEGF的分泌,激活ERK1/2和AP-1 DNA结合活性。血府逐瘀汤处理可减弱这种现象。此外,ERK1/2信号通路抑制剂U0126也可抑制VEGF的表达。
中医理论中的糖尿病血瘀证对调节ERK1/2-VEGF信号通路具有积极作用。活血化瘀中药可能是治疗DR的有效疗法。