法舒地尔在大鼠心肌缺血再灌注损伤模型中作用机制的研究.docVIP

法舒地尔在大鼠心肌缺血再灌注损伤模型中作用机制的研究 [摘要]目的观察Rho激酶在大鼠心肌细胞缺血再灌注损伤细胞凋亡中的作用，法舒地尔（fasudil，F）对缺血再灌注损伤细胞凋亡的影响。方法45只SD大鼠建立缺血再灌注损伤（IRI）模型，实验分3组：（1）空白对照组（C组）；（2）缺血再灌注+生理盐水组（IR组）；（3）缺血再灌注+法舒地尔组（FH组）。Western blot法检测肌球蛋白磷酸酶目标亚单位1（MYPT1）磷酸化水平，作为Rho激酶功能活化的标志，应用流式细胞仪检测心肌细胞的凋亡率。结果再灌注后MYPT1的磷酸化水平显著增加，IR组磷酸化MYPT1水平是正常对照组的3.66倍（P 0.01）。用法舒地尔干预后，FH组磷酸化MYPT1水平较IR组降低36.34%（正常对照组的2.33倍），FH组心肌细胞的凋亡率较IR组呈下降趋势（P 0.01）。结论Rho激酶在缺血再灌注心肌细胞中有促MYPT1磷酸化水平上调作用，法舒地尔可减少缺血再灌注心肌细胞凋亡的发生。 [关键词]法舒地尔；缺血再灌注损伤；细胞凋亡；大鼠 中图分类号：R542.2文献标识码：A文章编号：1009_816X（2014）05_0372_03 doi：10.3969/j.issn.1009_816x.2014.05.06Fasudil Mechanism on Rats with Myocardial Ischemia_reperfusion Injury. LU Yun， WU Li_yun， ZHAO Ming_hua， et al. Depatman of Medicament，Taizhou Integrated Traditional and Western Medicine Hospital， Zhejiang 317523，China [Abstract] Objective To observe the impact of Rho_kinase on rats’ cardiac myocyte apoptosis caused by ischemia_reperfusion injury and to investigate the protective effect of fasudil （F） against cardiac myocytes apoptosis in the course of ischemia_reperfusion injury. Methods Forty_five SD rats were randomly divided into 3 groups （n 15 each） to establish IPR model. （1） Group 1 were rats without injury serving as control （group C）； （2） Group 2 were rats suffered from ischemia_reperfusion with pretreatment of normal saline （group IR） or puerarin （group PI）. （3） Group 3 were rats suffered from ischemia_reperfusion but injected with fasudil （group FH）. The Western Blot analysis was employed to measure the phosphorylation level of myosin phosphatase target subunit 1（MYPT 1）， as the index of the Rho_kinase activity. The apoptosis ratio of rats cardiac myocytes was determined by flow cytometry. Results The injection resulted in a 3.66_fold increase in the phosphorylated MYPT1 level of group IR，as compared to group C （P 0.01）. After the interference of fasudil，the phosphorylated MYPT1 level of group FH is 36.34% lower than group IR （2.33 times compared to group C）. The apoptosis probability of cardiac myocytes in