中国中西医结合杂志2018，Vol.37Issue(11)：1334-1338,5.DOI:10.7661/j.cjim.20171010.235
通心络干预心肌微血管内皮细胞条件培养液对心肌细胞的保护作用
Protective Effects on Cardiomyocytes by the Conditioned Medium of Cardiac Microvascular Endothelial Cells Treated with Tongxinluo
摘要
Abstract
Objective To investigate the influence of conditioned medium of cardiac microvascular endothelial cells on cardiomyocytes and the protective effect of Tongxinluo. Methods Three kinds of conditioned cardiac microvascular endothelial cell conditioned medium were prepared : the normal cardiac microvascular endothelial cell-conditioned medium ( normal-conditioned medium, N-CM), the homocysteine-induced microvascular endothelial cell-conditioned medium ( homocysteine-conditioned medium,H-CM) and the cardiac microvascular endothelial cell-conditioned medium with Tongxinluo (Tongxinluo-conditioned medium,T-CM). The cardiomyocytes were randomly divided into four groups : the normal group, the N-CM group, the T-CM group and the H-CM group, corresponding conditioned medium were used to culture cardiomyocytes. The morphology of cardiomyocytes were observed with an inverted microscope, the contents of cardiac troponin T (cTnT) in cell supernatant were detected by enzyme-linked immunosorbent assay, the mitochondrial membrane potential (MMP) of the cells were detected by JC-1 reagent, and the protein expression levels of Cytochrome C (Cyt C) and calreticulin (CRT) were detected by Western blot. Results Compared with the normal group, the cells morphology, the contents of cTnT in cell supernatant, the MMP,and the protein expression levels of Cyt C and CRT did not significantly changed in the N-CM group (P ＞0. 05). Compared with N-CM group, the cells were shrinkaged,the cell edge was fuzzy, and there were a large number of dead cells floated in conditioned medium, the contents of cTnT in cell supernatant was increased, and the protein expression levels of Cyt C as well as CRT were up-regulated, while the MMP was decreased in the H-CM group(P ＜0. 01). Compared with HCM group, there was no obvious change in morphology, the cell edge was clear, and the dead cells floated in culture medium were decreased, the contents of cTnT in cell supernatant was decreased, and the protein expression levels of Cyt C as well as CRT were down-regulated,while MMP was increased in the H-CM group(P ＜0. 05, P ＜0. 01 ). Conclusion The cardiac microvascular endothelial cells injury can damage cardiomyocytes,and the underlying mechanism may be related to the damage of mitochondria by CRT, which can be inhibited by Tongxinlou.关键词
通心络/心肌微血管内皮细胞/心肌细胞/共培养/线粒体/钙网蛋白Key words
Tongxinluo/cardiac microvascular endothelial cells/cardiomyocytes/co-culture/mitochondrial membrane potential/calreticulin引用本文复制引用
位庚,常丽萍,尹玉洁,张月霞,梁小华,梁俊清..通心络干预心肌微血管内皮细胞条件培养液对心肌细胞的保护作用[J].中国中西医结合杂志,2018,37(11):1334-1338,5.基金项目
国家重点基础研究发展计划(No. 2012CB518600) （No. 2012CB518600）
石家庄市科学技术研究与发展计划项目(No. 161462343) （No. 161462343）
