摘要
目的探讨加味柴胡疏肝散及其拆方对心肌缺血合并抑郁症大鼠血管紧张素受体样受体(APJ)的内源性配体13(Apelin-13)、细胞外信号调节激酶(extracellular signal-regulated kinases,ERK)1/2的影响。方法SD雄性大鼠随机分为正常组、模型组、加味柴胡疏肝散组(11.7 g·kg^(-1))、拆方祛瘀化痰组(4.05 g·kg^(-1))、拆方疏肝行气组(3.15 g·kg^(-1))、拆方健脾养血组(4.5 g·kg^(-1))、氟西汀组(0.0018 g·kg^(-1))、曲美他嗪组(0.0054 g·kg^(-1))、辛伐他汀组(0.0018 g·kg^(-1))。除正常组外,其余各组大鼠均采用高脂饮食联合注射异丙肾上腺素(ISO)及慢性不可预见性温和应激(CUMS)方法建立高血脂状态的心肌缺血合并抑郁症大鼠模型,造模共8周。造模同时各药物组大鼠给予相应剂量的药物灌胃,正常组和模型组大鼠等体积的生理盐水灌胃,1次/d。采用旷场实验、糖水消耗实验评估大鼠焦虑抑郁状态;心脏B超检测左心室舒张末内径(LVDd)、左心室收缩末内径(LVDs)、左心室缩短分数(LVFS)、左心室射血分数(LVEF),HE染色观察心肌组织病理变化;采用酶标仪检测大鼠血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)含量;酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测大鼠血清和心脏组织Apelin-13含量;实时荧光定量聚合酶链式反应(real-time qPCR)、蛋白免疫印迹(Western blot)法检测大鼠心脏、海马组织ERK1/2的mRNA和蛋白表达。结果与正常组比较:模型组大鼠旷场实验水平运动距离和站立次数、糖水消耗率显著下降(P<0.01);心脏B超显示LVDd、LVDs显著增大,LVEF、LVFS显著下降(P<0.01),HE染色显示模型组大鼠心肌组织明显损伤;血清和心脏组织Apelin-13含量显著升高(P<0.01);海马ERK1、ERK2 mRNA及蛋白表达水平显著降低(P<0.01),心脏ERK1、ERK2mRNA及蛋白表达水平显著升高(P<0.01)。与模型组大鼠比较,加味柴胡疏肝散组及各拆方组上述指标的异常变化得到不同程度的改善(P<0.05或P<0.01),但全方组效果最佳。结论加味柴胡疏肝散及其拆方具有降血脂、抗抑郁、改善心肌损伤的作用,方中祛瘀化痰药、疏肝行气药、健脾养血药存在协同作用,全方可发挥最佳疗效,其机制可能与下调血清和心脏组织Apelin-13含量,调节心脏和海马组织ERK1/2的基因和蛋白表达有关。
Objective To explore the effects of Modified Chaihu Shugan Powder(加味柴胡疏肝散)and its decomposed prescriptions on the endogenous ligand 13(Apelin-13)and extracellularsignal-regulated kinases(ERK1/2)of angiotensin receptor-like receptor(APJ)in rats with myocardial ischemia and depression.Methods SD male rats were randomly divided into normal group,model group,Modified Chaihu Shugan Powder group(11.7 g·kg^(-1)),Quyu Huatan decomposed prescription group(4.05 g·kg^(-1)),Shugan Xingqi decomposed prescription group(3.15 g·kg^(-1)),Jianpi Yangxue decomposed prescription group(4.5 g·kg^(-1)),fluoxetine group(0.0018 g·kg^(-1)),trimetazidine group(0.0054 g·kg^(-1))and simvastatin group(0.0018 g·kg^(-1)).In addition to the normal group,the rats in other groups were treated with high-fat diet combined with isoproterenol(ISO)injection and chronic unpredictable mild stress(CUMS)to establish a rat model of myocardial ischemia with depression in hyperlipidemia state.The model was made for 8 weeks.At the same time,rats in each drug group were given the corresponding doses of drugs by gavage.Rats in the normal group and the model group were given the same volume of normal saline by gavage,once a day.The field experiment and the sugar water consumption experiment were used to assess the status of anxiety.The left ventricular end diastolic diameter(LVDd),left ventricular end systolic diameter(LVDs),left ventricular shortening fraction(LVFS)and left ventricular ejection fraction(LVEF)were detected by B-mode echocardiography,and the pathological changes of myocardium were observed by HE staining.The contents of total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C)and low density lipoprotein cholesterol(LDL-C)in serum of rats were detected by enzyme labeling instrument.Enzyme linked immunosorbent assay(ELISA)was used to detect the content of Apelin-13 in serum and heart tissue of rats.The mRNA and protein expression of ERK1/2 in heart and hippocampus of rats were detected by real-time quantitative polymerase chain reaction(real-time qPCR)and Western blot.Results Compared with those of the normal group,the horizontal movement distance,standing times and sugar water consumption rate of the model group were significantly decreased(P<0.01).Cardiac B-ultrasound showed that LVDd and LVDs were significantly increased,and LVEF and LVFS were significantly decreased(P<0.01).HE staining showed that the myocardial tissue of the model group was significantly damaged.The levels of Apelin-13 in serum and heart tissue were significantly increased(P<0.01);Hippocampal ERK1,ERK2 mRNA and protein expression levels were significantly reduced(P<0.01),and cardiac ERK1,ERK2 mRNA and protein expression levels were significantly increased(P<0.01).Compared with those of the model group,the abnormal changes of the above indicators in the Modified Chaihu Shugan Powder group and each decomposed group were improved to varying degrees(P<0.05 or P<0.01),but the whole prescription group had the best effect.Conclusion Modified Chaihu Shugan Powder and its decomposed prescriptions have the effects of lowering blood lipids,anti-depression and improving myocardial injury.This study reveals that there are synergistic effects of resolving blood stasis and phlegm,soothing liver and regulating Qi,strengthening spleen and nourishing blood drugs,which makes the whole prescription can exert the best effect.The mechanism may be related to the down-regulation of Apelin-13 levels in serum and heart tissue and the regulation of the gene and protein expression of ERK1/2 in the heart and hippocampus.
作者
蒋春建
李晓红
王茜
黄子娟
陆雅席
吴戈
杨苹
王呈祥
杨力强
孔鹏云
JIANG Chunjian;LI Xiaohong;WANG Qian;HUANG Zijuan;LU Yaxi;WU Ge;YANG Ping;WANG Chengxiang;YANG Liqiang;KONG Pengyun(Guangxi University of Chinese Medicine,Nanning 530200,Guangxi,China;Guangxi Key Laboratory of Chinese Medicine Foundation Research,Guangxi University of Chinese Medicine,Nanning 530200,Guangxi,China)
出处
《中华中医药学刊》
CAS
北大核心
2022年第12期171-177,I0056,共8页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(81560748,82060835)
广西自然科学基金(2018GXNSFAA050079)
广西中医基础研究重点实验室项目(19-245-14-06,19-050-45-07)
广西中医药大学非一流学科建设项目(2018XK018,2019XK054)
广西中医药大学校级重点研究生创新项目(YCSZ2020022)。
