摘要
目的观察电针预处理对脂多糖(LPS)诱导的脓毒症模型小鼠心功能障碍和炎性反应的影响,探讨电针预处理促心肌保护的可能机制。方法24只SPF级雄性C57BL/6J小鼠随机分为对照组、模型组和电针预处理组,每组8只。电针预处理组在异氟烷麻醉状态下电针双侧足三里穴,疏密波,频率2/15 Hz,强度2 mA,持续15 min;对照组、模型组同样的方式抓取、固定。电针结束后,模型组和电针预处理组腹腔注射LPS(10 mg/kg)构建脓毒症模型;对照组注射等量生理盐水。采用超声心动图评价心功能;ELISA法检测血清TNF-α、IL-1β水平;qPCR和Western blot法检测心肌组织中TNF-α、IL-1β、IL-6和IL-10 mRNA和蛋白表达量;流式细胞术检测心肌组织中F4/80+CD11b+总巨噬细胞、F4/80+CD11b+CD206lowM1型和F4/80+CD11b+CD206highM2型巨噬细胞含量。结果与对照组相比,模型组小鼠的LVEF、LVFS均显著下降(P<0.01),血清中TNF-α、IL-1β含量均升高(P<0.01),心肌组织中TNF-α、IL-1β、IL-6、IL-10 mRNA和蛋白表达增加(P<0.05,P<0.01),心肌组织中巨噬细胞含量增多(P<0.01),且M1型巨噬细胞比例增高(P<0.01);与模型组相比,电针预处理组的LVEF、LVFS均显著提高(P<0.01),血清中TNF-α、IL-1β含量下降(P<0.05,P<0.01),心肌组织中TNF-α、IL-1β、IL-6 mRNA和蛋白表达下降(P<0.05,P<0.01),而IL-10表达上升(P<0.05,P<0.01),巨噬细胞含量下降(P<0.01),且M2型巨噬细胞比例增高(P<0.01)。结论电针预处理可能通过促进心肌组织中巨噬细胞由促炎M1型向抗炎M2型极化,减轻全身和局部炎性反应水平,改善心功能,产生心肌保护效应。
OBJECTIVE To investigate the effects of electroacupuncture(EA)pretreatment on cardiac dysfunction and inflammatory reaction in lipopolysaccharide(LPS)induced sepsis mice,and to explore the possible mechanisms of EA preconditioning for cardioprotection.METHODS A total of 24 male C57BL/6J mice were randomized into 3 groups:control group,model group,and EA pretreatment group,with 8 mice in each group.Before modeling,mice in the EA pretreatment were given"Zusanli"(ST36)EA stimulation(2/15 Hz,2 mA,15 min)under anesthesia with isoflurane.The model group and the control group were only fixed for 15 min in the same way.The sepsis mice models were established by intraperitoneal injection of LPS(10 mg/kg).Similar to the control group,an equal volume of saline was administered.Cardiac function was measured by echocardiography and the contents of inflammatory cytokines in serum was examined by ELISA.The mRNA and protein expression levels of inflammatory cytokines in cardiac tissue were detected with qPCR and Western blot respectively,the number of F4/80+CD11b+macrophages and the contents of F4/80+CD11b+CD206low M1 and F4/80+CD11b+CD206high M2 macrophages in myocardium was determined by flow cytometry.RESULTS Compared with the control group,the LVEF,LVFS value of the model group was decreased(P<0.01),the serum levels of TNF-α,IL-1βincreased(P<0.01),the expression of TNF-α,IL-1β,IL-6,IL-10 mRNA and protein increased(P<0.01,P<0.05),the number of F4/80+CD11b+macrophages in cardiac tissue increased(P<0.01),the proportion of F4/80+CD11b+CD206low M1 macrophages increased(P<0.01).Nevertheless,EA pretreatment group could reverse these changes mentioned above in the model group.Compared with the model group,the LVEF,LVFS values increased significantly(P<0.01),the serum levels of TNF-α,IL-1βdecreased(P<0.05,P<0.01),the expression of pro-inflammatory cytokines TNF-α,IL-1β,IL-6,and the number of macrophages in the myocardium downregulated(P<0.05,P<0.01),the expression of anti-inflammatory interleukins IL-10(P<0.05)and the proport
作者
陈栎遥
杨文秀
施珺菁
杨嘉丽
傅淑平
于美玲
卢圣锋
CHEN Li-yao;YANG Wen-xiu;SHI Jun-jing;YANG Jia-li;FU Shu-ping;YU Mei-ling;LU Sheng-feng(Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing, 210023, China;Zhangjiagang Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Zhangjiagang, 215600, China)
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2021年第5期696-701,共6页
Journal of Nanjing University of Traditional Chinese Medicine
基金
国家自然科学基金(81774210,81574062)
江苏省第十五批“六大人才高峰”项目(YY-033)
江苏高校“青蓝工程”中青年学术带头人项目(2020年)
张家港市青年科技项目(ZJGQNKJ202014)。
关键词
电针预处理
脓毒症
心功能障碍
炎性反应
巨噬细胞极化
electroacupuncture preconditioning
sepsis
cardiac dysfunction
inflammatory response
macrophage polarization
