Diabetes Research Institute, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen; Geriatric Department, Zhongshan Hospital (Xiamen), Fudan University, Xiamen.
Diabetes Research Institute, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen.
Eur J Histochem. 2024 Nov 25;68(4):4140. doi: 10.4081/ejh.2024.4140.
Retinopathy is a common complication of diabetes mellitus and the leading cause of visual impairment. Danggui Buxue decoction (RRP) has been used as a traditional drug for the treatment of diabetic nephropathy for many years. The aim of this study was to investigate the effects of RRP on hypoxia-induced retinal Müller cell injury. A model of retinal Müller cell damage was created using high glucose levels (25 mmol/L) and/or exposure to low oxygen conditions (1% O2). RRP was given to rats by continuous gavage for 7 days to obtain drug-containing serum. After sterilization, the serum was added to the culture medium at a ratio of 10%. Cell viability, apoptosis, and cell proliferation were assessed using the CCK-8 kit, Annexin V-FITC/propidium iodide apoptosis kit, and EdU kit. The mRNA levels of angiogenesis factors (ANGPTL4, VEGF) and inflammatory factors (IL-1B, ICAM-1) were detected by RT-qPCR. Western blot analysis was employed to assess the levels of proteins related to the ATF4/CHOP pathway. Following hypoxia for 48 h and 72 h, there was a significant decrease in cell viability and proliferation, as well as a notable increase in apoptosis compared to the control group (21% O2). However, high glucose stimulation had no significant effect, and high glucose combined with hypoxia had no further damage to cells. After 48 h of exposure to low oxygen levels, the mRNA expression levels of ANGPTL4, VEGF, IL-1B, and ICAM-1 in retinal Müller cells were significantly higher than in the control group (21% O2). RRP treatment significantly alleviated the increase of cell apoptosis and the upregulation of IL-1B and-1 in retinal Müller cells induced by hypoxia. RRP has the potential to reduce the suppression of the ATF4/CHOP pathway in hypoxia-induced retinal Müller cells, and it significantly alleviates cell apoptosis through regulating inflammatory factors and the ATF4/CHOP pathway.
视网膜病变是糖尿病的常见并发症,也是导致视力损害的主要原因。当归补血汤(RRP)多年来一直被用作治疗糖尿病肾病的传统药物。本研究旨在探讨 RRP 对高糖和低氧诱导的视网膜 Müller 细胞损伤的作用。采用高糖(25mmol/L)和/或低氧(1%O2)条件建立视网膜 Müller 细胞损伤模型。用连续灌胃的方法给大鼠服用 RRP7 天,获得含药血清。血清经灭菌后按 10%的比例加入培养基。用 CCK-8 试剂盒、Annexin V-FITC/PI 凋亡试剂盒和 EdU 试剂盒检测细胞活力、凋亡和细胞增殖。用 RT-qPCR 检测血管生成因子(ANGPTL4、VEGF)和炎症因子(IL-1B、ICAM-1)的 mRNA 水平。用 Western blot 分析评估与 ATF4/CHOP 通路相关的蛋白水平。与对照组(21%O2)相比,缺氧 48h 和 72h 后,细胞活力和增殖明显下降,凋亡明显增加。然而,高糖刺激没有显著影响,高糖与低氧联合作用对细胞没有进一步的损伤。低氧 48h 后,视网膜 Müller 细胞中 ANGPTL4、VEGF、IL-1B 和 ICAM-1 的 mRNA 表达水平明显高于对照组(21%O2)。RRP 处理可显著减轻低氧诱导的视网膜 Müller 细胞凋亡增加和 IL-1B、-1 的上调。RRP 可减轻低氧诱导的视网膜 Müller 细胞中 ATF4/CHOP 通路的抑制,通过调节炎症因子和 ATF4/CHOP 通路显著减轻细胞凋亡。