高龄退行性心脏瓣膜病患者高迁移率族蛋白 B2 与瓣膜钙化的临床及实验研究

Clinical and Experimental Study of High Mobility Group Box-2 and Valvular Calcification in Elderly Patients with Degenerative Heart Valve Disease.

机构信息

Nantong University Medical School, Nantong, China,

Nantong University Medical School, Nantong, China.

出版信息

Cardiology. 2023;148(3):271-277. doi: 10.1159/000529973. Epub 2023 Mar 23.

DOI:10.1159/000529973

PMID:36958298

Abstract

INTRODUCTION

The aim of this study was to investigate the relationship between the high mobility group box-2 (HMGB2) and valve calcification in senile degenerative heart valve disease (SDHVD).

METHODS

According to the echocardiographic results, patients with calcified heart valves were used as the experimental group and patients without calcified heart valves were used as the control group; blood was drawn for testing, and serum levels of HMGB2 were measured by an enzyme-linked immunosorbent assay. Human heart valve interstitial cells (hVICs) cultured in vitro were randomly divided into two groups. The calcification group was cultured with a medium containing calcification induction solution and cells were induced on days 1, 3, and 5, and the control group was cultured with a standard medium. Expression of bone morphogenetic protein 4 (BMP-4) and HMGB2 in both groups was detected by Western blot. RT-PCR was performed to detect the expression of the HMGB2 gene during calcification. The hVICs were cultured in vitro for 4 days with different concentrations of exogenous HMGB2 (0.01 μg/mL, 0.1 μg/mL, 1 μg/mL, 2 μg/mL), while the control group was cultured with a standard medium and the expression of BMP-4 and NF-κB P65 was detected by Western blot.

RESULTS

The serum level of HMGB2 was 7.90 (5.92, 12.39) μg/L, higher than that of 7.06 (5.06, 9.73) μg/L in the valve calcification group in elderly patients with degenerative valve disease (p = 0.005); the differences were statistically significant. In in vitro experiments, the cellular calcification protein BMP-4 and the HMGB2 protein were higher in the calcification group compared to the control group (p < 0.05). Exogenous stimulation of hVICs with HMGB2 was able to upregulate the expression of BMP-4 and NF-κB P65 (p < 0.05).

CONCLUSIONS

HMGB2 is correlated with valvular calcification in senile degenerative heart valve disease. The HMGB2 protein may promote the process of SDHVD valve calcification by activating the NF-κB pathway and upregulating the expression of BMP-4.

摘要

简介

本研究旨在探讨高迁移率族蛋白 B2（HMGB2）与老年退行性心脏瓣膜病（SDHVD）中瓣膜钙化的关系。

方法

根据超声心动图结果，将钙化性心脏瓣膜患者作为实验组，无钙化性心脏瓣膜患者作为对照组；采集血液进行检测，采用酶联免疫吸附试验测定血清 HMGB2 水平。体外培养人心脏瓣膜间质细胞（hVICs），随机分为两组。钙化组用含钙化诱导液的培养基培养，分别在第 1、3、5 天诱导细胞，对照组用标准培养基培养。通过 Western blot 检测两组细胞骨形态发生蛋白 4（BMP-4）和 HMGB2 的表达。采用 RT-PCR 检测钙化过程中 HMGB2 基因的表达。体外培养 hVICs 4 天，分别用不同浓度的外源性 HMGB2（0.01μg/ml、0.1μg/ml、1μg/ml、2μg/ml）处理，对照组用标准培养基培养，通过 Western blot 检测 BMP-4 和 NF-κB P65 的表达。

结果

老年退行性瓣膜病患者中，HMGB2 血清水平为 7.90（5.92，12.39）μg/L，高于瓣膜钙化组的 7.06（5.06，9.73）μg/L（p=0.005），差异有统计学意义。体外实验中，钙化组细胞钙化蛋白 BMP-4 和 HMGB2 蛋白高于对照组（p＜0.05）。外源性刺激 hVICs 用 HMGB2 能上调 BMP-4 和 NF-κB P65 的表达（p＜0.05）。