黄连碱减轻脓毒症小鼠心肌损伤的机制  

Mechanism on the protective effect of coptisine against myocardial injury in sepsis mice

作　　者：周游 韩晓曼 雷一鸣 李婧[1] 杜佳敏 魏捷[2] 周晨亮[1] Zhou You;Han Xiaoman;Lei Yiming;Li Jing;Du Jiamin;Wei Jie;Zhou Chenliang(Department of Critical Care Medicine,the People's Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院重症医学科,湖北武汉430060 [2]武汉大学人民医院急诊科,湖北武汉430060

出　　处：《中国急救医学》2025年第4期341-348,共8页Chinese Journal of Critical Care Medicine

摘　　要：目的 探索黄连碱(COP)对脓毒症诱导小鼠心肌损伤的作用及机制。方法 将60只雄性C57BL/6小鼠随机分为假手术组(Sham组)、脓毒症组(CLP组)和治疗组(CLP+COP组),每组20只。CLP组和CLP+COP组小鼠通过盲肠结扎穿孔术(CLP)诱导脓毒症形成,Sham组则接受相同的手术操作,但不进行盲肠结扎和穿孔。CLP+COP组小鼠在术前3 d至术后4 d每日连续灌胃COP。术后7 d,连续记录3组小鼠的存活率。在术后7×24 h后,采用心脏超声评估各组小鼠的心脏功能。收集小鼠血液,检测血清中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)和心肌肌钙蛋白I(cTnI)的水平。采集各组小鼠心脏组织,进行苏木精-伊红(HE)染色、活性氧(ROS)染色及透射电子显微镜(TEM)检查。通过免疫印迹法(Western blot)检测心肌组织中凋亡相关蛋白(Bax、Bcl-2)和氧化应激相关通路蛋白(Nrf2、HO-1)的表达变化。结果 与Sham组比较,CLP组小鼠存活率明显下降,心功能严重受损,心肌细胞排列紊乱且稀疏,心肌损伤标志物和炎性因子水平显著升高,心肌线粒体结构受损,且组织中ROS水平明显升高(P均<0.05)。与CLP组比较,CLP+COP组小鼠存活率升高,但差异无统计学意义(P>0.05);COP可改善心功能及心肌病理改变,降低心肌损伤标志物和炎性因子水平,减轻心肌线粒体损伤并减少氧化应激反应(P均<0.05)。Western blot检测结果显示,与Sham组比较,CLP组小鼠心肌组织中Bax的表达水平明显升高,而Bcl-2、Nrf2和HO-1的表达水平明显降低(P均<0.05)。与CLP组比较,CLP+COP组小鼠的Bax表达水平降低,Bcl-2、Nrf2和HO-1的表达水平明显升高(P均<0.05)。结论 COP能减轻脓毒症小鼠心肌损伤,其机制可能与激活Nrf2/HO-1通路,减轻线粒体氧化应激和抑制细胞凋亡密切相关。Objective To explore the effects and mechanisms of coptisine(COP)on myocardial injury induced by sepsis in mice.Methods Sixty male C57BL/6 mice were randomly divided into three groups:Sham group,sepsis group(CLP group)and treatment group(CLP+COP group)with 20 mice in each group.In the CLP and CLP+COP groups,sepsis was induced by cecal ligation and puncture(CLP)surgery,while the Sham group underwent the same surgical procedure without CLP.Mice in the CLP+COP group were administered COP by daily gavage from 3 days prior to surgery until 4 days after surgery.The survival rate of the three groups was recorded daily for one week after surgery.After 7×24 h postoperatively,cardiac function was assessed by echocardiography.Blood samples were collected to measure the levels of interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),lactate dehydrogenase(LDH),creatine kinase-MB isoenzyme(CK-MB),and cardiac troponin I(cTnI)in serum.Subsequently,heart tissues from each group were collected for HE staining,reactive oxygen species(ROS)staining and transmission electron microscopy(TEM)analysis.Finally,Western blot was performed to examine the expression of apoptosis-related proteins(Bax,Bcl-2)and oxidative stress-related pathway proteins(Nrf2,HO-1)in myocardial tissues.Results Compared with the Sham group,the CLP group mice exhibited the significant decrease in survival rate,severe cardiac dysfunction,disorganized and sparse myocardial cell arrangement,a significant increase in myocardial injury markers and inflammatory factors,damaged mitochondrial structure,and a marked increase in ROS levels in the tissues(all P<0.05).Compared with CLP group,the survival rate in CLP+COP group increased,but there was no significant difference(P>0.05).COP effectively improved cardiac function and myocardial pathological changes,reduced myocardial injury markers and inflammatory factors,alleviated mitochondrial damage and decreased oxidative stress.Western blot results showed that the expression level of Bax in the myocard

关 键 词：黄连碱 脓毒症 脓毒症心肌损伤 线粒体氧化应激 细胞凋亡 核因子-E2相关因子2(Nrf2) 血红素加氧酶-1(HO-1) 

分 类 号：R28[医药卫生—中药学]