LI Xiao-ping, LI Lang, CHEN Rui-zhen, LIU Tang-wei, WU Wei-feng, SHEN E, YANG Ying-zhen, CHEN Hao-zhu
Key Laboratory of Viral Heart Disease, Ministry of Public Health, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Zhonghua Xin Xue Guan Bing Za Zhi. 2010 Sep;38(9):834-8.
in septic mice, myocardial calpain was activated and induced caspase-3 activation, the association between calpain activation and apoptosis was explored in this experiment.
in in vivo model, adult C57 mice were injected with lipopolysaccharide (LPS, 4 mg/kg, i.p.) to induce sepsis. Myocardial calpain and caspase-3 activities, protein levels of calpain-1, calpain-2, calpastatin, Bcl-2 and Bid were detected by Western blot analysis and myocardial apoptosis was detected by TUNEL, myocardiac function was evaluated by Langendorff system. In in vitro model, adult rat cardiomyocytes were incubated with LPS (1 microg/ml) or co-incubated with calpain inhibitor-III (10 micromol/L), calpain activity, caspase-3 activity, protein levels of Bcl-2 and Bid, and cardiomyocyte apoptosis were detected.
in septic mice, myocardial calpain and caspase-3 activity were increased up to 2.7- and 1.8-folds, respectively. Both calpain inhibitor-III and PD150606 significantly attenuated the increase of caspase-3 activity. Myocardial protein levels of calpain-1, calpain-2, calpastatin, Bcl-2 and Bid were similar between control and septic mice, and no cleavage of both Bcl-2 and Bid was found in septic mice. Calpain inhibitor-III significantly improved myocardial function in septic mice. In in vitro model, calpain and caspase-3 activities were increased after 4 h LPS treatment, co-treatment with calpain inhibitor-III prevented caspase-3 activity increase, protein Bcl-2 and Bid were similar between normal cardiomyocytes and LPS-treated cardiomyocytes. Cardiomyocyte apoptosis was similar in in vivo and in vitro septic models.
myocardial calpain activity is increased in LPS induced septic mice, subsequent caspase-3 activation may contribute to myocardial dysfunction in septic mice without aggravating myocardial apoptosis and Bcl-2 and Bid are not involved on calpain induced caspase-3 activation in our model.
在脓毒症小鼠中，心肌钙蛋白酶被激活并诱导半胱天冬酶-3激活，本实验探讨钙蛋白酶激活与细胞凋亡之间的关联。
在体内模型中，给成年C57小鼠腹腔注射脂多糖（LPS，4mg/kg）以诱导脓毒症。通过蛋白质免疫印迹分析检测心肌钙蛋白酶和半胱天冬酶-3活性、钙蛋白酶-1、钙蛋白酶-2、钙蛋白酶抑制蛋白、Bcl-2和Bid的蛋白水平，通过TUNEL检测心肌细胞凋亡，通过Langendorff系统评估心脏功能。在体外模型中，将成年大鼠心肌细胞与LPS（1μg/ml）孵育或与钙蛋白酶抑制剂III（10μmol/L）共同孵育，检测钙蛋白酶活性、半胱天冬酶-3活性、Bcl-2和Bid的蛋白水平以及心肌细胞凋亡。
在脓毒症小鼠中，心肌钙蛋白酶和半胱天冬酶-3活性分别增加至2.7倍和1.8倍。钙蛋白酶抑制剂III和PD150606均显著减弱了半胱天冬酶-3活性的增加。对照小鼠和脓毒症小鼠之间心肌钙蛋白酶-1、钙蛋白酶-2、钙蛋白酶抑制蛋白、Bcl-2和Bid的蛋白水平相似，且在脓毒症小鼠中未发现Bcl-2和Bid的裂解。钙蛋白酶抑制剂III显著改善了脓毒症小鼠的心脏功能。在体外模型中，LPS处理4小时后钙蛋白酶和半胱天冬酶-3活性增加，与钙蛋白酶抑制剂III共同处理可防止半胱天冬酶-3活性增加，正常心肌细胞和LPS处理的心肌细胞之间的蛋白Bcl-2和Bid相似。体内和体外脓毒症模型中的心肌细胞凋亡相似。
在LPS诱导的脓毒症小鼠中心肌钙蛋白酶活性增加，随后的半胱天冬酶-3激活可能导致脓毒症小鼠的心肌功能障碍，而不会加重心肌细胞凋亡，并且在我们的模型中Bcl-2和Bid不参与钙蛋白酶诱导的半胱天冬酶-3激活。
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