Renal denervation improves vascular endothelial dysfunction by inducing autophagy via AMPK/mTOR signaling activation in a rat model of type 2 diabetes mellitus with insulin resistance.

Abstract

Recent clinical and animal studies have shown that renal denervation (RDN) improves insulin sensitivity and endothelial dysfunction. However, the specific mechanism remains incompletely understood. The purpose of this study is to investigate the effects of RDN on endothelial dysfunction of type 2 diabetes mellitus (T2DM) rat models with insulin resistance and to explore the underlying molecular mechanisms. Male Sprague-Dawley rats were fed with or without high-fat diet allocated in different groups, combined with low-dose streptozotocin which induces a rat model to develop T2DM with insulin resistance. RDN was conducted 1week after the rat models fully developed T2DM. The animals were sub-divided into four groups randomly: control group (CON, n = 6), diabetic group (T2DM, n = 6), diabetic with sham surgery group (Sham, n = 6) and diabetic with RDN group (RDN, n = 6). Rats in all groups were studied at baseline, both preoperatively and 4weeks after RDN, respectively. Western blot was used to detect the expression of angiotensin-converting enzyme 2 (ACE2) protein and the expression of autophagy-related proteins Beclin1, LC3 and p62 and autophagy signaling pathway AMPK/mTOR proteins and apoptosis-related protein caspase-3 in the aorta endothelial cells. In addition, the effects of ACE2 on autophagy of human umbilical vein insulin resistance endothelial cell culture in vitro were also studied. RDN decreased plasma and renal tissue norepinephrine levels. The Von Willebrand factor level was also decreased, while the plasma level of nitric oxide (NO) was significantly increased after RDN. Compared with the T2DM group and the Sham group, the endothelium-dependent and endothelium-independent diastolic function of the RDN group was improved significantly, the expression of Beclin1, LC3, ACE2 and eNOS proteins was higher, and the level of p62 protein was decreased. Furthermore, we found that RDN can activate the expression of p-AMPK and inhibit the expression of p-mTOR. In cell culture experiment, ACE2 activated p-AMPK and inhibited p-mTOR, thus promoting autophagy. RDN may not only increase the expression of ACE2 in the vascular endothelium, but also can via ACE2 activate p-AMPK and inhibit p-mTOR, thus promoting autophagy and improving endothelial dysfunction.