基于CaMKⅡ/MPTP通路研究参附注射液改善脓毒症心肌损伤作用  

Research on the role of Shenfu Injection in improving sepsis-induced myocardial dysfunction via the CaMKⅡ/MPTP signaling pathway

作　　者：卢诗倩 向常清[1] 张世忠[2] 熊晓兰 张斌[1] 周立涛 黄伟[1] 赵发兵 姚卫 LU Shiqian;XIANG Changqing;ZHANG Shizhong;XIONG Xiaolan;ZHANG Bin;ZHOU Litao;HUANG Wei;ZHAO Fabing;YAO Wei(Department of Intensive Care Unit,Second People’s Hospital of China Three Gorges University/Yichang Second People’s Hospital,Hubei Yichang 443000,China;Basic Medical College of China Three Gorges University,Hubei Yichang 443002,China)

机构地区：[1]三峡大学附属第二人民医院/宜昌市第二人民医院重症医学科,湖北宜昌443000 [2]三峡大学基础医学院,湖北宜昌443002

出　　处：《中国医院药学杂志》2025年第7期770-776,共7页Chinese Journal of Hospital Pharmacy

基　　金：湖北省宜昌市医疗卫生科研项目(编号:A22-2-046)。

摘　　要：目的:研究参附注射液对脓毒症大鼠心肌损伤的保护作用,并研究钙/钙调蛋白依赖性蛋白激酶Ⅱ/线粒体通透性转换孔(CaMKⅡ/MPTP)参与其心肌保护机制。方法:将健康雄性SD大鼠随机分为假手术组(Sham)、盲肠结扎穿孔术致脓毒症心肌损伤模型组(CLP)、低剂量参附注射液组(4 mL·kg^(–1),LSF)、高剂量参附注射液组(10 mL·kg^(–1),HSF),每组16只。术后36 h分别观察SD大鼠脓毒症评分、存活率及腹腔解剖情况,HE染色观察各组心脏组织病理改变,超声心动图检测心功能,ELISA法测定血清TNF-α、IL-6、IL-10、cTnI、BNP水平、肌浆网Ca^(2+)-ATP酶(SERCA)活性,线粒体肿胀法测MPTP的开放程度,Western blot法检测大鼠心脏组织CaMKⅡ-δ、线粒体钙单向转运体(MCU)表达情况。结果:(1)术后36 h Sham组大鼠脓毒症评分、腹腔解剖情况及心脏组织病理改变正常,而CLP组大鼠情况明显变差,HSF组、LSH组较CLP组病变程度减轻,各组大鼠存活率分别为:Sham组100%、CLP组50%、HSF组88%、LSH组69%。(2)各组大鼠术后36 h显示:与Sham组相比,CLP组、HSF组和LSH组大鼠SV、LVEF、血清IL-10、SERCA都下降(P<0.05),HSF组和LSH组较CLP组升高(P<0.05)。CLP组、HSF组和LSH组较Sham组血清TNF-α、IL-6、cTnI、BNP、MPTP、CaMKⅡ-δ、MCU水平均升高(P<0.05),HSF组和LSH组较CLP组TNF-α、IL-6、cTnI、BNP、MPTP、CaMKⅡ-δ降低(P<0.05),且HSF组TNF-α、BNP、CaMKⅡ-δ较LSF组改善更明显(P<0.05),LSH组和HSF组MCU相比CLP组无显著改变(P>0.05)。结论:参附注射液可以改善脓毒症心肌损伤,CaMKⅡ/MPTP通路参与参附注射液对大鼠脓毒症心肌损伤的保护机制。OBJECTIVE To explore the protective effect of Shenfu Injection on rats with sepsis-induced myocardial dysfunction,and the cardioprotective effect of the calcium/calmodulin-dependent protein kinaseⅡand mitochondrial permeability transition pore(CaMKⅡ/MPTP)signaling pathway.METHODS Healthy male Sprague-Dawley(SD)rats were randomly divided into sham operation group,cecal ligation and puncture(CLP)group,and low-dose(LSF group),and high-dose(HSF group)Shenfu Injection group(4 mL·kg^(-1),and 10 mL·kg^(-1),respectively),with 16 in each group.At 36 hours postoperatively,the sepsis score,survival rate and abdominal anatomy of rats were observed.Pathological changes of rat cardiac tissue in each group were observed by hematoxylin and eosin(H&E)staining.Cardiac function was detected by echocardiography.Serum tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),interleukin-10(IL-10),cardiac troponin I(cTnI),B-type natriuretic peptide(BNP),and sarcoplasmic reticulum Ca^(2+)-ATPase(SERCA)were measured by enzyme-linked immunosorbent assay(ELISA).The degree of MPTP opening was measured by mitochondrial swelling method.Western blot was used to detect the protein expressions of CaMKⅡ-δ and mitochondrial calcium uniporter(MCU)in rat cardiac tissue.RESULTS At 36 hours postoperatively,the sepsis score,abdominal anatomy and cardiac histopathological changes of the Sham group were normal,while the conditions of the CLP group were significantly worse.The above lesions in CLP-induced rats treated with low-dose and high-dose Shenfu Injection were significantly milder than those of the CLP group.The 36-h survival rate was 100%,50%,88%and 69%in the sham operation group,CLP group,HSF group and LSF group,respectively.At 36 h postoperatively,stroke volume(SV)and left ventricular ejection fraction(LVEF),and serum IL-10 and SERCA were significantly reduced in the CLP,HSF and LSF groups than the sham operation group(P<0.05).They were significantly higher in the HSF and LSF group than the CLP group(P<0.05).Serum TNF-α,IL-6,cTnI,BNP,MP

关 键 词：钙/钙调蛋白依赖性蛋白激酶Ⅱ 线粒体通透性转换孔 参附注射液 脓毒症 心肌损伤 

分 类 号：R363[医药卫生—病理学] R96[医药卫生—基础医学]