乌头提取物通过调控蛋白激酶...及心肌线粒体呼吸功能的影响_郑曦.pdf

世界中西医结合杂志 2023 年第 18 卷第 1 期World Journal of Integrated Traditional and Western Medicine2023，Vol.18，No.1实验研究DOI:10 13935/j cnki sjzx 230110基金项目:2020 南充市市校科技战略合作项目(20SXQT0197)作者单位:南充市中心医院心血管内科，四川 南充 637000通信作者:王浩宇，Email:361536712 qq com乌头提取物通过调控蛋白激酶 C 信号通路对心力衰竭大鼠钙转运及心肌线粒体呼吸功能的影响郑曦陈世蓉熊挺淋王德伟王浩宇【摘要】目的探究乌头提取物通过调控蛋白激酶 C(Protein kinase C，PKC)信号通路对心力衰竭大鼠钙转运及心肌线粒体呼吸功能的影响。方法将 30 只雄性 SD 大鼠随机分为 3 组，假手术组 9 只、模型组 9 只和药物组 9 只(假手术组大鼠造模时死亡 1 只，造模组造模失败 1 只，死亡 1 只)。根据人临床应用剂量及大鼠换算系数折算给药剂量，药物组大鼠灌胃乌头提取液 0 04 mg/kg，其余两组大鼠均灌胃等量的生理盐水干预。均连续干预28 d 后，观察比较 3 组大鼠心功能指标左心室收缩压(Left ventricular systolic pressure，LVSP)，左心室舒张末压(Left ventricular end diastolic pressure，LVEDP);左室内压上升最大速率(LV+dp/dtmax);左室内压下降最大速率(LV dp/dtmax);T PC 检测心肌细胞中 DAG 、PKC mNA 和心肌组织中钙转运蛋白 mNA 表达;检测线粒体呼吸功能;蛋白质印迹检测线粒体呼吸链复合酶 的蛋白表达。结果模型组大鼠 LVSP、LV+dp/dtmax、LV dp/dtmax及心肌钙转运蛋白 y2、SECA2A mNA 表达较假手术组降低，LVEDP 及 DAG 、PKC mNA 表达较假手术组升高，差异有统计学意义(P 0 05);药物组大鼠 LVSP、LV+dp/dtmax、LV dp/dtma 及心肌钙转运蛋白 y2、SECA2A mNA 表达高于模型组，LVEDP 及 DAG 、PKC mNA 表达较模型组低，差异有统计学意义(P 0 05)。模型组大鼠态 3 呼吸、最大呼吸、C1、C2 值及线粒体呼吸链复合酶、表达较假手术组均降低，态 4 呼吸较假手术组升高，差异有统计学意义(P 0 05);模型组比较，药物组大鼠态 3 呼吸、最大呼吸、C1、C2 值及线粒体呼吸链复合酶、表达均升高，态 4 呼吸降低，差异有统计学意义(P 0 05)。结论乌头提取物能抑制 PCK 信号通路的激活，修复心室细胞钙转运，改善线粒体呼吸功能，进而修复心衰。【关键词】乌头提取物;PKC 信号通路;心力衰竭;钙转运;心肌线粒体呼吸功能【中图分类号】285 5【文献标识码】AEffects of Aconitum carmichaelii Extract on Calcium Transport and Myocardial Mi-tochondrial espiratory Function in ats with Heart Failure by egulating PKC Sig-naling PathwayZHENG Xi，CHEN Shi rong，XIONG Ting lin，WANG De wei，WANG Hao yu(Department of Cardiovascular Medicine，Nanchong Central Hospital，Nanchong Sichuan 637000)【Abstract】ObjectiveTo investigate the effects of Aconitum carmichaelii extract(ACE)on calcium transport andmyocardial mitochondrial respiratory function in rats with heart failure by regulating protein kinase C(PKC)signaling path-way MethodsThirty male SD rats were randomly divided into a sham operation group(n=9)，a model group(n=9)，and a drug intervention group(n=9)One rat in the sham operation group died during modeling，and in the model group，one rat failed to be modeled and one rat died The dosage was converted according to human clinical dosage and rat conver-sion factor The rats in the drug intervention group were intragastrically administered with ACE at 0 04 mg/kg，and those inthe other two groups were with the same amount of normal saline After continuous intervention for 28 days，the cardiacfunction indexes of the three groups were observed and compared The mNA expression of DAG and PKC in myo-cardial cells and calcium transporter in myocardial tissues was detected by T PC Mitochondrial respiratory functionwas detected Western blot was used to detect the protein expression of mitochondrial respiratory chain complexes esultsThe mNA expression of LVSP，LV+dp/dtmax，LV dp/dtmax，and cardiac calcium transporters y2 andSECA2A in the model group was lower than that in the sham operation group，and the mNA expression of LVEDP，DAG26世界中西医结合杂志 2023 年第 18 卷第 1 期World Journal of Integrated Traditional and Western Medicine2023，Vol.18，No.1，and PKC was higher than that in the sham operation group(P 0 05)The mNA expression of LVSP，LV+dp/dtmax，LV dp/dtmax，and cardiac calcium transporters y2 and SECA2A in the drug intervention group was higherthan that in the model group，and the mNA expression of LVEDP，DAG ，and PKC was lower than that in the mod-el group(P 0 05)The state respiration，maximal respiration，C1 value，C2 value，and the expression of mito-chondrial respiratory chain complexes and in the model group were lower than those in the sham operation group，andthe state respiration was higher than that in the sham operation group(P 0 05)Compared with the model group，thedrug intervention group showed increased state respiration，maximal respiration，C1 value，C2 value，and the ex-pression of mitochondrial respiratory chain complexes and，and decreased state respiration(P 0 05)Conclu-sionACE can inhibit the activation of the PCK signaling pathway，restore calcium transport in ventricular cells，and im-prove the mitochondrial respiratory function，thereby repairing heart failure【Keywords】Aconitum carmichaelii Extract;PKC Signaling Pathway;Heart Failure;Calcium Transport;MyocardialMitochondrial espiratory Function心力衰竭(Heart failure，HF)是全世界心血管领域的难题之一，近些年 HF 的发病率不断增加1，2。目前，临床在治疗心力衰竭上有很大的疗效，但其发病率和病死率仍然很高。HF 属中医学“心悸”“喘证”“水肿”“痰饮”等范畴3。中药乌头作为中医学中的温补药物，具有多种生物作用，例如祛风湿、止痛、麻醉等，尤其在镇痛方面效果显著，且无成瘾性，在麻醉止痛和抑制肿瘤细胞等方面效果较好4。文献显示，乌头对于心衰的治疗有较好的疗效，但目前其具体的作用机制尚不清楚5。蛋白激酶 C(Protein kinase C，PKC)属于丝氨酸/苏氨酸蛋白激酶，可通过诱导细胞分化、增殖而调节心室重构6。有研究发现，当心肌中穿膜钙流改变和钙离子稳态持续紊乱发生时，会导致病理性心室重构发生，PKC 激活可导致心脏功能的快速改变，进而对心室重构造成长期的影响7。目前，关于 PKC 对心力衰竭的作用机制尚未有明确报道，因此本研究旨在探究乌头提取物通过调控 PKC 信号通路对心力衰竭大鼠钙转运及心肌线粒体呼吸功能的影响。1材料与方法1 1实验材料1 1 1实验动物30 只雄性 SD 大鼠均购自三峡大学实验动物中心，许可证号:SCXK(鄂)2017 0012。体质量 200 220 g，SPF 级，自由饮水摄食，保持室内温度25 29，室内光照黑白交替 12 h 一循环。本研究已通过南充市中心医院医学伦理委员会批准 审批号:2021年审(043)号。1 1 2实验试剂乌头(来源于湖南省人民医院药剂科)，制备乌头提取液:取 300 g 乌头根，提取乙醇提取物浸膏，将其和水充分混匀呈悬液，萃取后用旋转蒸发仪回收溶剂，最终制取 3 g/ml 药物浓度的乌头提取物。1 1 3实验仪器Lepgen 96 实时定量 PC 仪(美国 Illumina 公司);Allegra 64 高速冷冻离