Huayu Tongmai Granules protects against vascular endothelial dysfunction via up-regulating miR-185 and down-regulating RAGE.

Xiaoming Liu,Xiaoni Yang,Dongli Wang,Lei Lei

doi:10.1042/bsr20180674

Abstract

Objective: Receptor of advanced glycation end products (RAGE) is a membrane protein that contributes to the initiation and progression of diabetic vascular complications, which is reported as a target of miR-185. Huayu Tongmai Granules is a Chinese herbal compound that is capable of treating diabetic angiopathy. The present study was designed to explore the molecular biological mechanism by which Huayu Tongmai Granules protects against diabetic angiopathy.Methods: The rat model of diabetes and hyperglucose cell model were established. The blood glucose was detected to verify whether the model was successfully established. Besides, serum nitric oxide (NO) and reactive oxygen species (ROS) concentrations of the rats in each group were determined. The quantitative real-time PCR analysis was performed to examine the mRNA expression levels of miR-185 and other miRNAs in femoral artery of rats or human umbilical vein endothelial cell line. Additionally, the protein levels of RAGE or Bax were determined using Western blotting. Cell apoptosis was determined by terminal dUTP nick-end labeling assay or flow cytometry.Results: In the present study, we found that Huayu Tongmai Granules significantly decreased blood glucose and serum ROS and up-regulated serum NO concentration. MiR-185 was low-expressed in diabetic rats; however, Huayu Tongmai Granules intervention up-regulated miR-185. Stable overexpression of miR-185 directly suppressed the expression of RAGE and further suppressed endothelial cell apoptosis.Conclusion: Huayu Tongmai Granules appears to have a therapeutic effect on diabetic angiopathy that is most probably mediated by miR-185/RAGE axis.

Highlights

Diabetes mellitus (DM) is a metabolic disorder referring to a disorder in the metabolism of carbohydrates, fats and proteins, which is defined by hyperglycemia, affecting over 400 million patients worldwide [1]
To identify the potential miRNAs that may be involved in the progression of diabetic angiopathy, we chose five previously reported lncRNAs and subjected them to quantitative real-time PCR (qRT-PCR) analysis to determine their expression during DM: miR-185, miR-92a, miR-712, miR-342-5p, and miR-320a
The results of terminal dUTP nick-end labeling (TUNEL) assay supported this fact that DM promoted the apoptosis of endothelial cells, while Tongmai Granules inhibited the apoptosis under high-glucose condition (Figure 2D, P

Summary

Introduction

Diabetes mellitus (DM) is a metabolic disorder referring to a disorder in the metabolism of carbohydrates, fats and proteins, which is defined by hyperglycemia, affecting over 400 million patients worldwide [1]. Accumulating evidence has strongly implied that diabetes has devastating effects on the vasculature leading to numerous vascular complications including cardiovascular diseases, retinopathy, neuropathy and nephropathy, which are the major causes of morbidity and mortality [2]. There is a growing evidence supporting that oxidative stress, collagen deposition, angiogenesis, and vascular remodeling are associated with the pathogenesis of diabetic vascular complications [3,4,5]. Emerging evidence has confirmed that diabetic macroangiopathy is characterized by vascular endothelial dysfunction and vascular fibrosis and cirrhosis [6].

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Conclusion