Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>

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-in...

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Main Authors: Xilin Ge, Caoxin Huang, Wenting Chen, Chen Yang, Wenfang Huang, Jia Li, Shuyu Yang
Format: Article
Language:English
Published: PAGEPress Publications 2024-11-01
Series:European Journal of Histochemistry
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Online Access:https://www.ejh.it/index.php/ejh/article/view/4140
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author Xilin Ge
Caoxin Huang
Wenting Chen
Chen Yang
Wenfang Huang
Jia Li
Shuyu Yang
author_facet Xilin Ge
Caoxin Huang
Wenting Chen
Chen Yang
Wenfang Huang
Jia Li
Shuyu Yang
author_sort Xilin Ge
collection DOAJ
description 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.
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publishDate 2024-11-01
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spelling doaj-art-28f75feea73a44da93708d60f688db1a2025-08-20T02:27:50ZengPAGEPress PublicationsEuropean Journal of Histochemistry1121-760X2038-83062024-11-0168410.4081/ejh.2024.4140Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>Xilin Ge0Caoxin Huang1Wenting Chen2Chen Yang3Wenfang Huang4Jia Li5Shuyu Yang6Diabetes Research Institute, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen; Geriatric Department, Zhongshan Hospital (Xiamen), Fudan University, XiamenDiabetes Research Institute, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, XiamenDiabetes Research Institute, The First Affiliated Hospital of Xiamen University, XiamenDiabetes Research Institute, The First Affiliated Hospital of Xiamen University, XiamenDiabetes Research Institute, The First Affiliated Hospital of Xiamen University, XiamenDiabetes Research Institute, The First Affiliated Hospital of Xiamen University, XiamenTraditional Chinese Medicine Studio, The First Affiliated Hospital of Xiamen University, Xiamen; Integrated Traditional Chinese and Western Medicine Center, School of Medicine, Xiamen University, Xiamen 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. https://www.ejh.it/index.php/ejh/article/view/4140Danggui Buxue decoctionhypoxiaretinal Müller cellsdiabetic retinopathyATF4/CHOP pathway
spellingShingle Xilin Ge
Caoxin Huang
Wenting Chen
Chen Yang
Wenfang Huang
Jia Li
Shuyu Yang
Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>
European Journal of Histochemistry
Danggui Buxue decoction
hypoxia
retinal Müller cells
diabetic retinopathy
ATF4/CHOP pathway
title Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>
title_full Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>
title_fullStr Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>
title_full_unstemmed Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>
title_short Effect of Danggui Buxue decoction on hypoxia-induced injury of retinal Müller cells <i>in vitro</i>
title_sort effect of danggui buxue decoction on hypoxia induced injury of retinal muller cells i in vitro i
topic Danggui Buxue decoction
hypoxia
retinal Müller cells
diabetic retinopathy
ATF4/CHOP pathway
url https://www.ejh.it/index.php/ejh/article/view/4140
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