细胞凋亡参与脑心综合征心肌损伤及针刺干预作用

目的观察心肌细胞凋亡在脑心综合征(cerebral-cardiac syndrome,CCS)发病中的作用机制及针刺的干预作用。方法从60只健康SD大鼠中随机选择10只作为伪手术组,其余50只用于CCS模型复制,从模型复制成功的大鼠中随机选取30只,分成模型组、针刺组和非经非穴组,每组10只。采用胶原酶加肝素联合注射大鼠尾状核方法复制CCS模型,伪手术组向大鼠尾状核注入等量生理盐水。模型复制后72h处死动物,分别采用苏木精-伊红(hematoxylin-eosin,HE)染色、脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling,TUNEL)和免疫组织化学法检测心肌组织病理变化、细胞凋亡及半胱氨酸天冬氨酸蛋白酶3(Caspase 3)的表达。结果 HE染色示伪手术组心肌结构正常,模型组心肌细胞变性、坏死,大量平滑肌断裂;TUNEL及免疫组织化学染色提示模型组出现较多凋亡细胞,Caspase 3大量表达,针刺组TUNEL染色阳性细胞和Caspase 3阳性细胞均较模型组和非经非穴组显著减少(P<0.01)。结论心肌细胞凋亡参与了CCS大鼠心肌损伤,其机制可能与上调凋亡蛋白酶Caspase 3表达有关,而针刺可拮抗CCS大鼠心肌组织细胞凋亡。

Cell Apoptosis in Myocardial Damage and Therapeutic Effect of Acupuncture among Rats with Cerebral-Cardiac Syndrome

Objective To investigate the mechanism of cardiomyocyte apoptosis in the development of cerebral-cardiac syndrome (CCS) and the therapeutic effect of acupuncture. Methods Ten rats were randomly selected from 60 healthy Sprague-Dawley rats as sham-operation group; a CCS model was induced in the other 50 rats. Thirty rats were randomly selected from the rats with CCS and equally divided into model group, acupuncture group, and non-acupoint group. The CCS model was induced by injection of collagenase and heparin into the caudate nucleus, while the rats in sham-operation group were injected with an equal volume of saline into the caudate nucleus. These rats were sacrificed at 72 h after the CCS model was established. The pathological changes, cell apoptosis, and Caspase-3 expression in myocardial tissue were evaluated by hematoxylin-eosin (HE) staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and immunohistochemistry. Results The sham-operation group had normal myocardial structure, while the model group had myocardial degeneration and necrosis, as well as extensive smooth muscle rupture, according to the HE staining. The TUNEL and immunohistochemical staining showed that cell apoptosis and Caspase-3 expression were frequently seen in the model group, and the acupuncture group had significantly decreased TUNEL-positive cells and Caspase-3-positive cells compared with the model group and non-acupoint group (P<0.01). Conclusion Cardiomyocyte apoptosis is involved in myocardial damage among rats with CCS, which may be due to the up-regulated Caspase-3 expression. Acupuncture can suppress the cardiomyocyte apoptosis in rats with CCS.