Hu-Zhang-Qing-Mai-Yin Inhibits Proliferation of Human Retinal Capillary Endothelial Cells Exposed to High Glucose

Yuan-Yuan Yu,Hong Zhang,Pei An,Qiu-Ping Liu,Xin Luan,Meng-Ting Li,Chao Lv,Yu-Ying Chen

doi:10.3389/fphar.2021.732655

Abstract

Background: Diabetic retinopathy (DR) is one of the serious complications of diabetes and an important cause of blindness. Despite much research on the pathogenesis of DR, there is still a lack of safe and effective treatment methods. Hu-zhang-qing-mai-yin (HZQMY), a Chinese medicine formula, has been clinically used in the safe and effective treatment of DR for many years. However, the systematic pharmacological research is lacking. The aim of this study was to evaluate the anti-DR effects of HZQMY and explore the possible mechanism involved. Methods: The constituents of HZQMY were analyzed by LC-MS/MS. DR model was established by high glucose simulation on human retinal capillary endothelial cells (HRCECs) in vitro. The cell viability, cell proliferation, cell apoptosis, and tube formation were assessed. Subsequently the related mechanisms were analyzed by assays for JC-1 mitochondrial membrane potential (MMP), intracellular ROS, ATP, western blot and proteomics. Results: 27 main chemical components contained in HZQMY were identified. HZQMY significantly inhibited the viability and proliferation of HRCECs exposed to high glucose, and promoted the apoptosis. In addition, HZQMY also boosted the release of ROS and suppressed tube formation of HRCECs under high glucose exposure. Meanwhile, HRCECs treated with high glucose released more ROS than normal cells, which could be markedly inhibited by HZQMY in a dose-dependent manner. Additionally, western blot assay indicated that HZQMY increased the expression of proteins related to the P38 signaling pathway and inhibited nuclear factor kappa-B (NF-κB) pathway. Proteomic analysis predicted that HSPA4, MAPK3, ENO1, EEF2 and ERPS may be the candidate targets of HZQMY in HRCECs. Conclusions: HZQMY inhibited the proliferation and promoted the Mitochondria related apoptosis of HRCECs exposed to high glucose possibly through regulating P38 and NF-κB signaling pathway.

Highlights

Diabetes is one of the most common chronic metabolic diseases, with an estimated 592 million diabetics worldwide by 2035 (Duh et al, 2017)
50–150 μg/ml HZQMY significantly reduced the cell viability increased by high glucose after 24 h incubation (Figures 2A,B), and the inhibitory effect was more pronounced at 48 h (Figures 2C,D)
These results suggested that high glucose can improve cell viability, while HZQMY can reverse the promotion effect in a time and dose-dependent manner

Summary

Introduction

Diabetes is one of the most common chronic metabolic diseases, with an estimated 592 million diabetics worldwide by 2035 (Duh et al, 2017). The main treatment methods for DR include laser therapy, intravitreal injection of steroid hormones or taking anti-VEGF drugs, oral administration of calcium hydroxybenzene sulfonate, etc., but there are problems of ineffective or accompanied by a large number of side effects (Ihnat et al, 2007). It is an urgent need for finding the therapy with good safety and effectivity. HZQMY significantly inhibited the viability and proliferation of HRCECs exposed to high glucose, and promoted the apoptosis. Conclusions: HZQMY inhibited the proliferation and promoted the Mitochondria related apoptosis of HRCECs exposed to high glucose possibly through regulating P38 and NF-κB signaling pathway

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Results

Conclusion