中国动脉硬化杂志2012，Vol.20Issue(4)：321-326,6.
脂联素通过减轻内质网应激介导的凋亡抑制心肌细胞钙化
Adiponectin Inhibited Calcification in Cultured Rat Neonatal Cardiomyocytes via Reducing Endoplasmic Reticulum Stress-mediated Apoptosis
赵旭静 1边云飞 1白瑞 1王敏 1肖传实2
作者信息
- 1. 山西医科大学第二临床医学院心内科,山西省太原市 030001
- 2. 山西医科大学第一临床医学院心内科,山西省太原市 030001
- 折叠
摘要
Abstract
Aim To investigate the effects of adiponectin on calcification in cultured cardiomyocytes via reducing endoplasmic reticulum stress-mediated apoptosis in vitro. Methods Primary cultured cardiomyocytes were obtained from neonatal rats by collagenase II digestion method. The ot-actin expression as molecular marker of the cardiomyocytes was observed by immunocytochemistry. The cells cultured for 72 h were used in the experiment and divided into groups randomly; control group, calcified myocardial cells (CMC) group, adiponectin + CMC (3, 10, 20, 30 mg/L) groups. The morphological changes of the eardiomyocytes were observed under phase contracted microscope. CMC was prepared by incubation with p-glycerophosphate ( BGP). Calcification was confirmed by Von Kossa staining and Alizarin Red S. Calcium content and alkaline phosphatases (ALP). Alkaline phosphatase was measured by spectrophotometric measurement of p-nitorphenol release. Osteocalcin was detected by radioinununoassay. The content of lactate dehydrogenase (LDH) was measured. The cardiomyocyte apoptosis was detected by flow cytometry. The expression levels of glucose regulated protein 78 (GRP78)/cysteine-containing aspartate-specific proteases-12 (Caspase-12)/osteoprotegerin (OPG) mRNA were detected by quantitative real-time polymerase chain reaction ( qRT-PCR). Results The immunocyto-
chemical stain for smooth muscle a-actin confirmed cardiomyocytes phenotype and the raulticellular nodules spontaneouslyappeared in cardiomyocytes culture. ALP activity and the content of calcium and osteocalcin in calcified myocardial cells was increased by 138% and 136.9% (all P <0.05), respectively, compared with control. The apoptotic rate and the levels of LDH was also significantly increased. Compared to untreated cells, the mRNA levels of GRF78 and Caspase-12 increased in CMC, but the mRNA levels of OPG decreased. Adiponectin elevated the level of OPG mRNA, but suppressed ALP activity, the content of calcium, osteocalcin secretion and decreased mRNA levels of CRP78 and Caspase-12 in a dose-dependent manner. Conclusion Adiponectin inhibits calcification in cultured rat neonatal cardiomyocytes via reducing endoplasmic reticulum stress-induced apoptosis and plays a protective role by extenuation of cardiomyocytes calcification.关键词
脂联素/钙化/内质网应激/心肌细胞/凋亡Key words
Adiponectin/Calcification/Cardiomyocytes/Endoplasmic Reticulum Stress/Apoptosis分类
医药卫生引用本文复制引用
赵旭静,边云飞,白瑞,王敏,肖传实..脂联素通过减轻内质网应激介导的凋亡抑制心肌细胞钙化[J].中国动脉硬化杂志,2012,20(4):321-326,6.
