摘要
目的 观察电针刺激对慢性心力衰竭(CHF)大鼠模型及心肌纤维化细胞模型的影响,探讨电针对CHF的治疗作用机制.方法 选取24 只雄性Wistar大鼠随机分为4 组,分别为假手术组、假手术+电针组、心肌纤维化组及心肌纤维化+电针组,每组6 只.将H9c2 大鼠心肌细胞随机分为4 组,对照组、对照+电针组、血管紧张素Ⅱ处理组及血管紧张素Ⅱ处理+电针组.心肌纤维化组大鼠采用腹主动脉缩窄法建立CHF模型,假手术组大鼠仅分离腹主动脉不进行钳夹,心肌纤维化+电针组大鼠在心肌纤维化组的基础上联合电针刺激大鼠双侧"内关"穴干预,假手术+电针组大鼠在假手术组的基础上联合电针刺激大鼠双侧"内关"穴干预.血管紧张素Ⅱ处理组采用含血管紧张素Ⅱ的培养基对H9c2 大鼠心肌细胞进行培养及传代,对照组H9c2 大鼠心肌细胞在正常培养基内进行培养及传代,血管紧张素Ⅱ处理+电针组是血管紧张素Ⅱ处理组的基础上在培养皿中予以电刺激,对照+电针组在对照组的基础上在培养皿中予以电刺激.检测各组大鼠干预12、16、20 周室间隔舒张末期厚度(IVSd)、左室舒张末期后壁厚度(LVPWd)、左室收缩末期内径(LVEDs)、左室舒张末期内径(LVEDd)及左室射血分数(LVEF),以及血清B型钠尿肽(BNP)、乳酸脱氢酶(LDH)、肌酸激酶(CK)水平,检测各组H9c2 大鼠心肌细胞在干预12、16、20 周重组蛋白基质金属蛋白酶 9(MMP9)及基质金属蛋白酶抑制剂-1(TIMP-1)相对表达情况.结果 与假手术组干预同期比较,心肌纤维化组干预12、16、20 周时IVSd、LVP-Wd、LVEDs及LVEDd均升高(P<0.05),LVEF降低(P<0.05),但假手术+电针组干预12、16、20 周时各项指标与假手术组干预同期比较差异均无统计学意义(P>0.05).与心肌纤维化组干预同期比较,心肌纤维化+电针组干预 12、16、20 周时IVSd、LVPWd、LVEDs及LVEDd均降低(P<0.05),LVEF升高(P<0.05).与假手术组干预同期比较,心肌纤维化组干预12、16、20 周时BNP、LDH及CK水平均升高(P<0.05),但假手术+电针组干预 12、16、20 周时各项指标与假手术组干预同期比较差异均无统计学意义(P>0.05).与心肌纤维化组干预同期比较,心肌纤维化+电针组干预 12、16、20 周时BNP、LDH及CK水平均降低(P<0.05).与对照组干预同期比较,血管紧张素Ⅱ处理组干预12、16、20 周时MMP-9 及TIMI-1 相对表达水平均升高(P<0.05),但对照+电针组干预12、16、20 周时各项指标与对照组干预同期比较差异均无统计学意义(P>0.05).与血管紧张素Ⅱ处理组干预同期比较,血管紧张素Ⅱ处理+电针组干预 12、16、20 周时MMP-9 及TIMI-1 相对表达水平均降低(P<0.05).结论 电针对CHF的治疗作用可能是通过抑制心室重构,改善心功能,保护心肌细胞,减轻心肌纤维化而实现.
Abstract
Objective To observe the effects of electroacupuncture(EA)on chronic heart failure(CHF)rat model in vivo and myocardial fibrosis model in vitro,and to explore the therapeutic mechanism of electroacupuncture for CHF.Methods Twenty-four male Wistar rats were randomly divided into 4 sham operation group,sham operation + EA group,myocardial fibrosis group,and myocardial fibrosis + EA group,with 6 rats in each group.Transverse aortic coarctation was performed for CHF model-ing in rats,and those in sham operation group were managed by i-solation of the abdominal aorta without clamping.Electroacupunc-ture at the Neiguan acupoint on bilateral sides was performed.Rat cardiomyocyte cell line H9c2 was induced with blank control,control + EA,angiotensin Ⅱ,and angiotensin Ⅱ + EA.H9c2 cells induced with blank control were routinely cultured and passaged.Angiotensin Ⅱ and electric stimulations were intervened in vitro.At 12,16 and 20 weeks,rat interventricular septal thickness(IVSd),left ventricular posterior wall thickness(LVPWd),left ventricular end-systolic dimension(LVEDs),left ventricular end-diastolic dimension(LVEDd)and left ventricular ejection fraction(LVEF)were recorded.Serum B-type na-triuretic peptide(BNP),lactate dehydrogenase(LDH)and creatine kinase(CK)were measured.At 12,16 and 20 weeks,relative levels of matrix metalloproteinase 9(MMP9)and tissue inhibitor of metalloproteinases 1(TIMP-1)in H9c2 cells were detected.Results Compared with rats of sham operation group,those in myocardial fibrosis group had significantly higher IVSd,LVPWd,LVEDs and LVEDd at 12,16 and 20 weeks,but lower LVEF(P<0.05).There were no significant differences in IVSd,LVPWd,LVEDs,LVEDd and LVEF between sham operation group and sham operation + EA group(P>0.05).Compared with rats of myocardial fibrosis group,those in myocardial fibrosis + EA group had significantly lower IVSd,LVPWd,LVEDs and LVEDd at 12,16 and 20 weeks,but higher LVEF(P<0.05).Compared with rats of sham operation group,those in myocardial fibrosis group had significantly higher BNP,LDH and CK levels at 12,16 and 20 weeks(P<0.05).There were no significant differences in BNP,LDH and CK between sham operation group and sham operation + EA group(P>0.05).Compared with rats of myocardial fibrosis group,those in myocardial fibrosis + EA group had significantly lower BNP,LDH and CK(P<0.05).Compared with H9c2 cells induced with blank control,significantly upregulated MMP-9 and TIMI-1 were detected in angiotensin Ⅱ-induced cells at 12,16 and 20 weeks(P<0.05).No significant differences were detected in the expression levels of MMP-9 and TIMI-1 between H9c2 cells induced with blank control and control + EA(P>0.05).Com-pared with H9c2 cells induced with angiotensinⅡ,treatment of angiotensinⅡand EA significantly downregulated MMP-9 and TIMI-1at 12,16 and20 weeks(P<0.05).Conclusion The therapeutic effect of EA on CHF is attributed to the inhibition of ventricular remodeling,improvement of cardiac function,protection of myocardial cells,and alleviation of myocardial fibrosis.
基金项目
新疆维吾尔自治区自然科学基金(2021D01C81)
新疆维吾尔自治区人民医院院内项目(20210118)
国家科技期刊平台
NSTL
万方数据