摘要
观察氧化型低密度脂蛋白(ox-LDL)对原代培养的乳鼠心肌细胞钙敏感受体(CaSR)的表达和细胞凋亡的影响。将原代培养的乳鼠心肌细胞随机分为2组:正常对照组(control group)和ox-LDL组(ox-LDL group)。用MTT比色法、电镜、TUNEL染色和流式细胞计数仪等方法观察心肌细胞凋亡;用Western blot观察CaSR、Cytc、Bax、caspase-3和Bcl-2的蛋白表达;用激光共聚焦显微镜观察细胞内游离钙浓度的变化。与正常组比较,ox-LDL组:CaSR表达呈时间与剂量依赖性上调,细胞内钙增加,细胞凋亡增加,Cytc、Bax和caspase-3表达升高,细胞存活率和Bcl-2表达降低。ox-LDL可使原代培养心肌细胞的CaSR表达增加,通过激活线粒体通路,促进细胞凋亡。
To observe effect of oxidized low density lipoprotein （ox-LDL） on calcium-sensing receptors expression and apoptosis in culture myocardial cells from neonatal rats. The primary cultured cardiac cells of neonate rat were divided into two groups randomly：the control group and ox-LDL group. The survival rate of cell and apoptosis was measured by MTT chromatometry,electromicroscope,TUNEL staining and flow cytometry assay; the expressions of CaSR,cytochrome c （cyt-c）,Bax,Bcl-2 and caspase-3 protein were analyzed using Western blot; the intracellular calcium concentration was measured using a laser scanning confocal microscope. Compared with the control group,the expression of CaSR,cyt-c,Bax and caspase-3 were up-regulated,the expression of Bcl-2 and cell viability were down-regulated,the apoptotic ratio and intracellular calcium concentration were increased in ox-LDL group. Especially,the increase of CaSR expression in ox-LDL group was in time-and dose-dependent manner. ox-LDL can increase the expression of CaSR in the primary cultured cardiac cells of neonate rat,which promote apoptosis by activating mitochondria pathway.
基金
高等学校博士学科点专项科研基金资助项目(20070226012)
国家自然科学基金资助项目(30871012
30470688
81000059和30700288)
黑龙江省卫生厅科研课题资助项目(2009-214)
哈尔滨医科大学研究生创新基金资助项目(HCXB2008006)
黑龙江省研究生创新科研资金资助项目(YJSCX2008-106HLJ)
关键词
钙敏感受体
心肌细胞
氧化型低密度脂蛋白
细胞凋亡
calcium-sensing receptors
myocardial cells
oxidized low density lipoprotein
apoptosis
