Janus激酶-信号转导子与转录激活子通路介导粒细胞集落刺激因子影响冠状动脉微栓塞后心肌细胞凋亡的实验研究  被引量:11

Granulocyte colony stimulating factor attenuated myocardial apoptosis via Janus kinase 2/ signal transducer and activator of transcription signal transduction pathway in rats with coronary microembolization

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作  者:张飞龙[1] 陈良龙[1] 李淑梅[2] 王伟伟[1] 

机构地区:[1]福建医科大学附属协和医院心内科,福州350001 [2]福州总医院476临床部

出  处:《中华心血管病杂志》2008年第3期254-259,共6页Chinese Journal of Cardiology

基  金:福建省卫生厅青年基金立项课题资助项目(2005-2-16)

摘  要:目的探讨粒细胞集落刺激因子(G-CSF)对大鼠冠状动脉微栓塞(CME)后心肌细胞凋亡的影响以及Janus激酶-信号转导子与转录激活子(JAK/STAT)通路的介导作用。方法将92只雄性成年SD大鼠,随机分成CME组(24只)、G-CSF组(24只)、JAK2特异性抑制剂(AG490)组(G-CSF+AG490,24只)和假手术组(20只)。CME组、G-CSF组及AG490组升主动脉夹闭后自左室腔内注入自体微血栓,造成CME,假手术组注入等量生理盐水。G-CSF组及AG490组术后2h起给予皮下注射重组人G-CSF(rhG-CSF)100μg·kg^-1·d^-1持续5d,AG490组同时给予AG490溶液腹腔注射5mg·kg^-1·d^-1,其他组给予等量生理盐水。术后3d、1、2及4周处死动物。各组心肌样品中以实时定量聚合酶链式反应法检测Bcl-2、Bax、Fas及FasL的mRNA表达,并计算Bcl-2/Bax比值;以Western blot法检测Caspase-3、裂解多聚二磷酸腺苷-核糖聚合酶(PARP)、总JAK2(t-JAK2)、磷酸化JAK2(p-JAK2)、t-STAT3以及p-STAT3蛋白的表达;脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法测定凋亡细胞。结果(1)与假手术组比较。CME组术后Bcl-2、Bax、Fas及FasL的mRNA表达均升高,Bcl-2/Bax比值降低(0.28±0.04比2.98±0.49),Caspase-3(0.762±0.129比0.133±0.027)及PARP蛋白(0.992±0.146比0.386±0.074)表达增强,心肌细胞凋亡指数升高(P〈0.05或P〈0.01);t-JAK2、p-JAK2、t-STAT3以及p-STAT3蛋白表达差异无统计学意义。(2)与CME组比较,G-CSF组术后p-JAK2与p-STAT3蛋白表达明显增强,Bax、Fas及FasL的mRNA表达明显减弱,Bcl-2的mRNA表达增强,Bcl-2/Bax比值升高(2.07±0.29比0.28±0.04),Caspase-3(0.371±0.041比0.762±0.129)及PARP蛋白(0.548±0.093比0.992±0.146)表达减弱;心肌细胞凋亡指数下降(P〈0.05或P〈0.01);t-JAK2及t-STAT3蛋白表达则差异无统计Objective To investigate the effects of granulecyte colony stimulating factor (G-CSF) on myocardial apoptosis following coronary microembolization (CME) and possible role of Janus kinase/ singnal transducer and activator of transcription (JAK/STAT) pathway in this process. Methods A total of 92 male Sprague Dawley rats were randomized into CME ( n = 24), G-CSF ( 100μg · kg^-1·d^-1 i.p. 2 hours post CME for5 days, n=24), JAK2 inhibitor AG490 (G-CSF plus AG490, 5 mg · kg^-1·d^-1i. p. 2 hours post CME for 5 days, n = 24 ), all rats received left ventricular injection of homologous microthrombotic particle suspension post clamping the ascending aorta. Sham-operated group ( n = 20 )served as control. The rats were sacrificed at day 3,7,14 and 28 after operation. The myocardial mRNA expressions of Bcl-2, Bax, Fas, FasL and GAPDH which was used as the intracomparison, were evaluated by real time PCR. The ratio of Bel-2/Bax was compared. The protein expression of Caspase-3, cleaved PARP, t-JAK2, p-JAK2, t-STAT3 and p-STAT3 were detected by western blot. Myocardial apeptesis were examined by TUNEL staining. Results Compared with Sham rats, the mRNA of Bcl-2, Bax, Fas and FasL significantly increased whereas the ratio of Bcl-2/Bax(0.28 ± 0.04 vs. 2.98 ± 0.49 ) significantly decreased and the protein expression of Caspase-3 (0.762 ±0. 129 vs. 0. 133 ±0.027), PARP(0. 992 ±0. 146 vs. 0. 386 ±0. 074)and the myocardial apeptesis index( 17.2 ± 1.9 vs. 1.2 ± 0. 6) significantly increased in CME hearts ( all P 〈 0.05 ). rhG-CSF significantly attenuated CME induced changes and cotreatment with JAK2 inhibitor AG490 abolished the effects of rhG-CSF. The protein expressions of t-JAK2 and t-STAT3 among the groups were similar. P-JAK2 and p-STAT3 protein expressions were significantly increased in G-CSF group compared to other groups ( P 〈 0.05 ). Conclusion G-CSF attenuated myocardial apeptosis induced by CME via JAK2/STAT3 pathway.

关 键 词:冠状动脉疾病 栓塞 粒细胞集落刺激因子 细胞凋亡 信号传导 

分 类 号:R686[医药卫生—骨科学]

 

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