摘要
目的探讨法舒地尔对盐敏感性高血压大鼠心室重构及心肌细胞凋亡的影响。方法本实验于2016年2月—2017年6月在陕西省医学实验动物中心实验室完成。选取6周龄健康雄性Dahl盐敏感大鼠69只,采用随机数字表法分为生理盐组、高盐组及法舒地尔组,每组23只。生理盐组大鼠给予含盐量为0.3%的正常饲料饲养;高盐组大鼠给予含盐量为8%的特制饲料喂养,自由饮水;法舒地尔组大鼠给予含盐量为8%的特制饲料喂养,自由饮水,建模成功后6 h腹腔注射法舒地尔,连续治疗1周。药物治疗结束后1周采用高分辨小动物超声成像系统检测大鼠左心室舒张末期内径(LVEDd)、左心室收缩末期内径(LVESd)、左心室短轴缩短率(LVFS)、左心室射血分数(LVEF),并测量前壁厚度、后壁厚度;HE染色观察心肌细胞形态;VG染色法测算胶原容积积分;TUNEL法检测心肌细胞凋亡率,实时荧光定量反转录-聚合酶链反应(RT-PCR)检测RhoA激酶mRNA相对表达量,Western blot法检测RhoA激酶相对表达量。结果 (1)药物治疗结束后1周,生理盐组和法舒地尔组大鼠LVEDd、LVESd小于高盐组,LVFS和LVEF高于高盐组,前壁厚度和后壁厚度大于高盐组(P<0.05);法舒地尔组大鼠LVEDd、LVESd、前壁厚度及后壁厚度小于生理盐组,LVFS和LVEF高于生理盐组(P<0.05)。(2)生理盐组大鼠心肌细胞排列有序,心肌间质无炎性细胞浸润及纤维化瘢痕形成;高盐组大鼠心肌细胞排列紊乱,巨噬细胞浸润明显增多同时伴有明显纤维化重构;与高盐组相比,法舒地尔组大鼠尽管有炎性细胞浸润,但心肌细胞排列有序且无纤维化瘢痕组织形成,心肌纤维化重构较轻。(3)药物治疗结束后1周,生理盐组和法舒地尔组大鼠胶原容积积分低于高盐组,法舒地尔组大鼠胶原容积积分高于生理盐组(P<0.05)。(4)药物治疗结束后1周,生理盐组和法舒地尔组大鼠心肌细胞凋亡率低于高盐组,法舒地尔组大鼠心肌细胞凋亡率高于生理盐组(P<0.05)。(5)药物治疗结束后1周,生理盐组和法舒地尔组大鼠RhoA激酶mRNA和RhoA激酶相对表达量低于高盐组,法舒地尔组大鼠激酶mRNA和RhoA激酶相对表达量高于生理盐组(P<0.05)。结论法舒地尔能有效抑制盐敏感性高血压大鼠心室重构及心肌细胞凋亡,其机制可能与降低RhoA激酶表达有关。
Objective To investigate the impact of fasudil on myocardial reconstruction and apoptosis of myocardial cells in rats with salt-sensitive hypertension. Methods This experiment was completed in the Central Laboratory of Shaanxi Medical Laboratory Animal Center from February 2016 to June 2017. A total of 69 healthy male Dahl salt-sensitive rats（ 6 week-age） were selected and divided into A group,B group and C group according to random number table,each of 23 rats. Rats in A group received normal feed with 0. 3% salt content; rats in B group received special feed with 8% salt content and free drinking; rats in C group received special feed with 8% salt content and free drinking,and then received intraperitoneal injection of fasudil for 1 week after successful modeling. After 1 week of medical treatment, High Resolution Small Animal Ultrasound Imaging System was used to detect the LVEDd,LVESd,LVFS,LVEF,anterior wall thickness and posterior wall thickness;HE staining method was used to observe the myocardial cellular morphology,VG staining method was used to measure and calculate collagen volume fraction,TUNEL method was used to detect the apoptotic rate of myocardial cells,RT-PCR was used to detect the relative expression quantity of RhoA kinase mRNA,Western blot method was used to detect the relative expression quantity of RhoA kinase. Results（ 1） After 1 week of medical treatment,LVEDd and LVESd in A group and C group were statistically significantly smaller than those in B group,LVFS and LVEF in A group and C group were statistically significantly higher than those in B group,while anterior wall thickness and posterior wall thickness in A group and C group were statistically significantly larger than those in B group（ P〈0.05）; LVEDd,LVESd,anterior wall thickness and posterior wall thickness in C group were statistically significantly smaller than those in A group, while LVFS and LVEF in C group were statistically significantly higher than those in B group（ P〈0.05）.（ 2） In A group,myocardial cells aligned orderly,without inflammatory cell infiltration or fibrotic scar formation in myocardial interstitium; in B group,myocardial cells aligned mussily,with obvious increased macrophage infiltration and fibrous remodeling; compared with B group,myocardial cells aligned orderly with little inflammatory cell infiltration but without fibrotic scar formation in C group,the severity of myocardial fibrosis was milder.（ 3）After 1 week of medical treatment,collagen volume fraction in A group and C group was statistically significantly lower than that in B group,respectively,while collagen volume fraction in C group was statistically significantly higher than that in A group（ P〈0.05）.（ 4） After 1 week of medical treatment,apoptotic rate of myocardial cells in A group and C group was statistically significantly lower than that in B group, respectively, while apoptotic rate of myocardial cells in C group was statistically significantly higher than that in A group（ P〈0.05）.（ 5） After 1 week of medical treatment,relative expression quantity of RhoA kinase mRNA and RhoA kinase in A group and C group was statistically significantly lower than that in B group,respectively,while relative expression quantity of RhoA kinase mRNA and RhoA kinase in C group was statistically significantly higher than that in A group,respectively（ P〈0.05）. Conclusion Fasudil can effectively inhibit the myocardial reconstruction and apoptosis of myocardial cells in rats with salt-sensitive hypertension,the action mechanism may mostly correlate with down-regulation of RhoA kinase.
出处
《实用心脑肺血管病杂志》
2017年第12期49-54,共6页
Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease
基金
国家自然科学基金资助项目(81660079)
陕西省自然科学基础研究计划项目(2016JM8123)
