Arterial Calcification Is Regulated Via an miR-204/DNMT3a Regulatory Circuit Both In Vitro and in Female Mice

Abstract

Arterial calcification is a common cardiovascular disease that initiates from a process of osteoblastic differentiation of vascular smooth muscle cells (VSMCs). Accumulating evidence has demonstrated that microRNAs play an important role in regulating arterial calcification. miR-204 was significantly downregulated in calcified human renal arteries from patients with uremia; calcified arteries of mice, due to 5/6 nephrectomy with a high-phosphate diet (5/6 NTP); and in VSMCs induced by high phosphate concentration. The overexpression of miR-204 alleviated the osteoblastic differentiation of VSMCs. Bisulphite sequencing PCR revealed that CpG sites upstream of miR-204 DNA were hypermethylated in calcified VSMCs; in calcified arteries of mice, due to 5/6 NTP; and in calcified renal artery tissues from patients with uremia. Moreover, increased DNMT3a resulted in the hypermethylation of miR-204 in high phosphate concentration-induced VSMCs, whereas 5-aza-2'-deoxycytidine could restore the expression of miR-204 in high phosphate concentration-induced VSMCs. Moreover, we found that DNMT3a was the target of miR-204, and the methylation ratio of miR-204 was decreased significantly, meaning that the expression of miR-204 was restored when DNMT3a was knocked down by using DNMT3a small interfering RNA, resulting in abrogation of the effect of high phosphate concentration on VSMC calcification. The progress of arterial calcification is regulated by the miR-204/DNMT3a regulatory circuit.

Plasma exosomes derived from patients with end-stage renal disease and renal transplant recipients have different effects on vascular calcification

“…The expression of proteins was determined by Western Blot as previously described [3]. Brie y, the concentration of protein was detected using a BCA kit (Beyotime, Shanghai, China).…”

“…Vascular calci cation is mainly characterized as the medial of aortic calci cation (termed as Mönckeberg's calci cation), and it often happens in patients with chronic kidney disease, especially end stage renal disease (ESRD) [1,2]. Lots of studies have demonstrated that vascular calci cation is not a simple passive process of calcium and phosphorus deposition but an active regulatory process, and the transdifferentiation of vascular smooth muscle cells (VSMCs) into osteoblast-like cells is regarded to be the key pathophysiological factor of vascular calci cation [3][4][5]. However, the speci c mechanism of vascular calci cation in patients with ESRD is still not entirely elucidated.…”

Melatonin alleviates vascular calcification and ageing through exosomal miR‐204/miR‐211 cluster in a paracrine manner

“…Mice were fed with 12‐hour daylight/darkness transition in Animal House of the Second Xiang‐Ya Hospital. To generate vascular calcification and ageing in mice, the 5/6 NTP mouse model was established following the procedure previously described . Briefly, 6‐week‐old C57BL/6 mice were treated with an intraperitoneal injection of pentobarbital sodium (50 mg/kg) to induce general anaesthesia before operation.…”

“…vascular calcification and ageing in mice, the 5/6 NTP mouse model was established following the procedure previously described. 28 Briefly, 6-week-old C57BL/6 mice were treated with an intraperitoneal injection of pentobarbital sodium (50 mg/kg) to induce general anaesthesia before operation. The anaesthesia state of the mice was validated frequently by loss of the pedal withdrawal reflex.…”

The Epigenetic Landscape of Vascular Calcification: An Integrative Perspective

“…Among them, 15 (40.5%) have been confirmed or are suspected to promote or aggravate VC in different scenarios, while 22 (59.5%) protect against VC formation or progression. Direct gene targets have been identified for 14 miRNAs with negative VC regulatory capacity, including miR-29a/29b [14], miR-29b-3p [34], miR-30b [16,47], miR-30c [16], miR-30e [25], miR-34b/34c [29,64], miR-125b [13,22] miR-133a [19], miR-135a [30], miR-182 [42], miR-204 [15,41,50], and miR-205 [21], while few direct targets have been identified for miRNAs with VC enhancement ability (miR-34a [39], miR-128-3p [48], and miR-135a-3p [17]). In the following sections, we describe the clinical features of each VC-regulating miRNA based on their propensity for positive or negative vascular influences.…”