摘要
目的:检测大鼠心肌梗死(心梗)后心肌组织中转化生长因子β诱导基因-22(TSC-22)蛋白水平表达的动态变化,并在细胞水平上研究白细胞介素-6(IL-6)是否对其蛋白表达有诱导作用。方法:雄性Sprague-Dawley大鼠,通过结扎左冠状动脉前降支制备心梗模型。分为心肌梗死模型组(心梗组)和假手术对照组(假手术组),于术后1天、3天、7天、2周、4周进行心脏标本取材,酶联免疫吸附(ELISA)法检测大鼠左心室心肌组织中TSC-22蛋白的表达。胰酶消化法分离培养乳鼠原代心肌细胞,随机分为心肌细胞对照组和心肌细胞实验组,心肌细胞对照组正常培养,心肌细胞实验组分别在2.5、5.0、10.0 ng/ml浓度的IL-6刺激下培养24 h。采用ELISA法、免疫细胞化学法和免疫印迹法(Western blot)等方法检测TSC-22蛋白表达。结果:ELISA法结果表明,大鼠心肌梗死1天、7天心梗组比假手术组左心室心肌组织中TSC-22蛋白含量显著升高,4周时则显著降低,差异均有统计学意义(P均<0.05)。心肌细胞实验的ELISA法结果表明,在10.0 ng/ml的IL-6刺激24 h后,心肌细胞实验组比心肌细胞对照组TSC-22蛋白表达水平显著升高,差异有统计学意义(P<0.05),在2.5、5.0 ng/ml IL-6刺激24 h后,心肌细胞实验组与心肌细胞对照组比较差异无统计学意义;免疫细胞化学法和蛋白免疫印迹法结果显示,10.0 ng/ml的IL-6刺激24 h后,TSC-22在细胞核中的染色明显加深。结论:大鼠心梗后的急性期,如梗死1天、7天后,左心室心肌组织中TSC-22蛋白水平迅速升高。炎症细胞因子IL-6可能是其在心梗后上调的诱导因素之一。
Objective ：To investigate the protein expression of transforming growth factor-β stimulated clone-22 （TSC-22）in rats＇ infarcted myocardium ,and to study the effect of interleukin-6（ IL-6 ）on TSC-22 protein expression in primary cultured cardiomyocytes.
Methods ：①The myocardial infarction（MI） model was created hy left anterior descending coronary artery ligation ,and the rats were divided into MI group and Sham operation group. The animals were sacrificed 1,3,7 days and 2,4 weeks post-infarction, TSC-22 protein expression in left ventricular myocardium was examined by EHSA.②Primary cultured cardiomyocytes were obtained by enzymatic digestion method from nsonatal rats. The cells were divided into two groups. IL-6 group, the cardiomyocytes were treated with IL-6 at 2. 5 ng/ml,5 ng/ml,and 10 ng/ml respectively for 24 hours,and Control group,the cells were cultured without IL-6. TSC-22 protein expression in cardiomyocytes was measured by ELIZA ,immunocyloelaemistry and Western blot analysis.
Results：①TSC-22 protein expression in left ventricular myocardium in MI group was increased than that in Sham operation group 1 day and 7 days post-infaretion by ELISA,while it was decreased in MI group 4 weeks post-infarction,P〈0. 05 respectively. ②TSC-22 protein expression in 10 ng/ml of IL-6 treated eadiomyocytes was increased than that in Contral group cells by ELISA,P〈0. 05 ,meanwhile,the elevated expression was also presented by immunocytochemistry and Western blot analysis respectively. There were no statistic changes of TSC-22 protein levels between 2.5 ng/ml,5 ng/ml of IL-6 treated cardiomyocytes and the cells in Control group.
Conclusion： In the acute phase of MI （〈7 days）, the myocardium TSC-22 protein expression was immediately elevated in rats,the cytokine IL-6 might be one of the regulators for such modulation.
出处
《中国循环杂志》
CSCD
北大核心
2011年第3期227-230,共4页
Chinese Circulation Journal
基金
科技部"973"项目课题(2010CB529506)
"重大新药创制"科技重大专项--<创新药物临床研究技术平台建设>(2008ZX09312-018)
