加味柴胡疏肝散对心肌缺血合并抑郁症大鼠CaMKⅡ/CREB信号通路的影响  

Effect of Modified Chaihu Shugansan on CaMKⅡ/CREB Signaling Pathway in Rats with Myocardial Ischemia and Depression

作　　者：万芬 李晓红[1,2] 陈营 潘杨钰 罗艳娜 卢芳歌 郑春成[1] 孔鹏云 王呈祥 杨力强[1,2] WAN Fen;LI Xiaohong;CHEN Ying;PAN Yangyu;LUO Yanna;LU Fangge;ZHENG Chuncheng;KONG Pengyun;WANG Chengxiang;YANG Liqiang(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Key Laboratory of Chinese Medicine Foundation Research,Guangxi University of Chinese Medicine,Nanning 530200,China;Shanxi University of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]广西中医药大学,南宁530200 [2]广西中医药大学广西中医基础研究重点实验室,南宁530200 [3]山西中医药大学,晋中030619

出　　处：《中国实验方剂学杂志》2025年第9期1-11,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广西青年基金项目(2021GXNSFBA196059);广西自然科学基金重点项目(2022GXNSFDA035086);国家自然科学基金项目(82060835)。

摘　　要：目的:观察加味柴胡疏肝散对心肌缺血合并抑郁症模型大鼠海马和心脏组织钙调蛋白依赖性激酶Ⅱ型(CaMKⅡ)/环磷腺苷反应元件结合蛋白(CREB)信号通路的影响,探讨该方防治冠心病合并抑郁症的作用机制。方法:采用高脂饮食加腹腔注射异丙肾上腺素(ISO)联合慢性不可预见性温和应激(CUMS)制备心肌缺血合并抑郁症模型,将108只SD雄性大鼠随机分为正常组,模型组,加味柴胡疏肝散高、中、低(23.4、11.7、5.85 g·kg^(-1))剂量组,CaMKⅡ抑制剂(KN93)组,KN93+加味柴胡疏肝散高、中、低剂量组,每组12只。造模第1天至造模结束,每天给药,每天1次。在第7、8周,KN93组、KN93+加味柴胡疏肝散高、中、低剂量组大鼠每周腹腔注射3次KN93。第8周造模结束对各组大鼠进行糖水偏好、旷场、高架行为学测试;观察各组大鼠心电图Ⅱ导联变化;苏木素-伊红(HE)染色观察各组大鼠心脏组织变化;酶标仪检测大鼠血清中甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)含量和乳酸脱氢酶(LDH)水平;紫外分光光度计检测血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)水平;酶联免疫吸附测定法(ELISA)检测大鼠血清中单核细胞趋化蛋白-1(MCP-1)含量;实时荧光定量聚合酶链式反应(Real-time PCR)法检测海马和心脏组织CaMKⅡ、CREB mRNA表达;蛋白免疫印迹法(Western blot)检测海马和心脏组织CaMKⅡ、磷酸化(p)-CaMKⅡ、CREB、p-CREB的蛋白表达。结果:与正常组比较,模型组大鼠糖水偏好率、旷场活动总距离、进入旷场中心区域次数和进入高架十字开放臂次数比例显著降低(P<0.01),心电图见ST段弓背样抬高,HE染色见心肌纤维严重变性,排列紊乱,大量炎细胞浸润,血清中TC、LDL含量升高(P<0.01),HDL含量显著降低(P<0.01),CK、CK-MB、LDH水平和MCP-1含量明显升高(P<0.05,P<0.01),海马组织中CaMKⅡ、CREB mRNA和p-CaMKⅡ、p-CREB蛋白表达明显Objective:To observe the effects of modified Chaihu Shugansan on the calmodulin-dependent protein kinaseⅡ(CaMKⅡ)/cAMP-response element binding protein(CREB)signaling pathway in the hippocampus and heart tissue of a rat model with myocardial ischemia and depression and explore the mechanism by which this formula prevents and treats coronary heart disease combined with depression.Methods:The model of myocardial ischemia combined with depression was established by high-fat diet,intraperitoneal injection of isoproterenol(ISO),and chronic unpredictable mild stress(CUMS).A total of 108 SD male rats were randomly divided into normal group,model group,high(23.4 g·kg^(-1)),medium(11.7 g·kg^(-1)),and low(5.85 g·kg^(-1))dose groups of modified Chaihu Shugansan,CaMKⅡinhibitor(KN93)group,and KN93+high,medium,and low dose groups of modified Chaihu Shugansan,with 12 rats in each group.From the first day of modeling to the end of modeling,drugs were administered once a day.In the seventh and eighth weeks,the KN93 group and the KN93+high,medium,and low dose groups of modified Chaihu Shugansan were intraperitoneally injected with KN93 three times weekly.At the end of the eighth week,behavioral tests including sucrose preference,open field,and elevated plus maze were conducted.Electrocardiogram(ECG)leadⅡchanges were observed in each group of rats,and hematoxylin-eosin(HE)staining was performed to observe changes in heart tissue.Serum levels of triglycerides(TG),total cholesterol(TC),high-density lipoprotein(HDL),low-density lipoprotein(LDL),and lactate dehydrogenase(LDH)were measured by using an enzyme-labeled instrument.Creatine kinase(CK)and creatine kinase-MB(CK-MB)were detected by ultraviolet spectrophotometry,while serum monocyte chemoattractant protein-1(MCP-1)was measured by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect mRNA expression of CaMKⅡand CREB in hippocampal and heart tissue,and Western blot was performed to assess

关 键 词：加味柴胡疏肝散 心肌缺血合并抑郁症 钙调蛋白依赖性激酶Ⅱ型(CaMKⅡ)/cAMP反应元件结合蛋白(CREB)信号通路 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R259