心肌钙反常模型细胞因子的表达及地尔硫卓的干预作用

目的探讨心肌钙反常模型诱导心肌细胞因子的表达及地尔硫卓的干预作用。方法采用Langendorff装置对离体大鼠心脏进行钙反常灌流(无钙灌流液5min,立即换正常钙灌流液灌流30min),建立心肌细胞钙超载模型。设立钙超载组、正常对照组、无钙对照组、钙超载给药组(地尔硫卓)和无钙给药组。电镜和光镜观察心肌病理和超微结构的变化。原子分光光度计检测心肌钙离子浓度,RT-PCR法测心肌细胞因子TNF-α、IL-1β、IL-6、TGF-β1和IL-10的表达。结果钙超载组光镜下未见明显炎症细胞浸润,电镜结果显示心肌细胞膜、细胞核和线粒体明显破坏。与正常对照组比较,TNF-α、IL-1β和IL-6明显增高,而TGF-β1和IL-10无明显变化。钙超载给药组(地尔硫卓)显示心肌损伤明显改善,TNF-α、IL-1β和IL-6明显下降。正常对照组、无钙对照组和无钙给药组心肌光镜和电镜下无明显差异,心肌Ca2+浓度差异无统计学意义,细胞因子的表达差异也无统计学意义。结论心肌钙反常模型诱导心肌表达TNF-α、IL-1β和IL-6,对TGF-β1和IL-10无明显影响,地尔硫卓能抑制心肌细胞钙超载诱导的TNF-α、IL-1β和IL-6表达。

Diltiazem inhibits cytokine expression in rat myocardium of calcium paradox model

Aim To explore the cardiac cytokine expression in rat model of myocardial calcium overload, and the intervention from diltiazem.Methods The intracellular Ca~(2+) overload was induced by the isolated rat heart subjected to 5 min Ca~(2+) depletion and 30 min Ca~(2+) repletion (Ca~(2+) paradox) by the Langendorff technique.There were five groups in this study, including Ca~(2+) overload group, normal control group, Ca~(2+) depletion control group, Ca~(2+) overload-diltiazem group, and Ca~(2+) depletion-diltiazem group.The views of myocardial pathology and ultrastruction were observed by electron microscope and light microscope respectively. The cardiac intracellular Ca~(2+)]_i was detected by atom spectrophotometer. The expression of TNF-α, IL-1β, L-6, TGF-β1, and IL-10 was detected by RT-PCR method.Results In Ca~(2+) overload group, few inflammatory cells were found in myocardium under the light microscope. And the views of electron microscope presented that cardiocyte membranes, nucleolus, and mitochondria were disorganized obviously.Compared with normal control group, the inflammatory cytokines as TNF-α, IL-1β, and IL-6 were increased significantly whereas there was nearly no difference of the expression of TGF-β1 and IL-10 in Ca~(2+) overload group.Ca~(2+) overload-diltiazem group showed that TNF-α, IL-1β, and IL-6 were decreased significantly. There were no statistical differences in the structure of myocardium, intracellular Ca~(2+)]_i, and cardiac cytokines expressions in the three control groups, including normal control group, Ca~(2+) depletion control group and Ca~(2+) depletion-diltiazem group.Conclusions Instead of TGF-β1 and IL-10, the expression of TNF-α, IL-1β, and IL-6 is increased obviously in myocardium of calcium paradox model. Diltiazem can inhibit the cardiac expression of TNF-α, IL-1β, and IL-6 induced by myocardial calcium overload.