中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2014年
2期
151-156
,共6页
秦延军%张新亮%于悦卿%卞晓华%董士民
秦延軍%張新亮%于悅卿%卞曉華%董士民
진연군%장신량%우열경%변효화%동사민
脓毒症%心肌损伤%凋亡%促红细胞生成素%线粒体细胞膜电位%核转录因子%炎症因子%大鼠
膿毒癥%心肌損傷%凋亡%促紅細胞生成素%線粒體細胞膜電位%覈轉錄因子%炎癥因子%大鼠
농독증%심기손상%조망%촉홍세포생성소%선립체세포막전위%핵전록인자%염증인자%대서
Sepsis%Myocardial injury%Apoptosis%Chondriosome membrane potential%Nuclear factor κB p65%Erythropoietin%Inflammatory cytokines%Rat
目的 探讨促红细胞生成素(EPO)对脓毒症大鼠心肌损伤的保护作用及其机制.方法 通过盲肠结扎穿孔引流术(CLP)建立脓毒症大鼠模型.健康SD大鼠96只,随机(随机数字法)分为3组:假手术组(Sham组,n=32)、脓毒症组(CLP组,n=32)、EPO组(n=32,脓毒症模型基础上EPO 1000 IU/kg腹腔内注射).分别于造模术后3、6、12、24 h记录心脏血流动力学指标变化,测定血清炎症反应因子水平及心肌酶变化,应用流式细胞仪检测心肌细胞线粒体膜电位、心肌细胞凋亡及心肌组织核因子-κB (NF-κB p65)表达水平；光镜下观察大鼠心肌细胞的病理学变化.所有数据以均数±标准差(-x±s)表示,采用SPSS 13.0统计软件分析,组间比较采用t检验,以P ＜0.05为差异具有统计学意义.结果 (1)与Sham组相比,CLP组大鼠左室收缩压(LVSP)、左室舒张末压(LVEDP)、左室压力上升最大速度(+dp/dtmax)、左室压力下降最大速度(-dp/dtmax)均有不同程度恶化(P ＜0.05,P＜0.01),EPO组各指标降低幅度小于CLP组(P＜0.05,P＜0.01)；(2)与Sham组相比,CLP组各时间点血清肿瘤坏死因子-α(TNF-α)、白介素-6 (IL-6)、C-反应蛋白水平(CRP)、肌钙蛋白I(cTNI)、肌酸激酶(CK)、乳酸脱氢酶(LDH)和谷草转氨酶(AST)水平及心肌组织NF-κB p65表达水平等指标均有不同程度升高(P＜0.05),而EPO组上述各指标低于CLP组(P＜0.05),与之相反,EPO组血清抗炎因子白介素-10 (IL-10)各时间点均高于CLP组；(3)与Sham组相比,CLP组心肌细胞凋亡率升高(P＜0.01)；(4)心肌细胞线粒体膜电位较Sham组降低(P＜0.01),与CLP组相比,EPO组心肌细胞凋亡率明显降低,线粒体膜电位升高(P＜0.叭)；(5)应用EPO干预后,心脏组织损害的病理改变较脓毒症组减轻.结论 脓毒症大鼠中,外源性EPO可阻抑心肌细胞线粒体膜电位降低,抑制心肌细胞凋亡；同时抑制NF-κB活化,减少促炎因子生成,减轻心脏组织的炎症损伤.以上两种机制发挥保护受损心肌、改善心功能的作用.
目的 探討促紅細胞生成素(EPO)對膿毒癥大鼠心肌損傷的保護作用及其機製.方法 通過盲腸結扎穿孔引流術(CLP)建立膿毒癥大鼠模型.健康SD大鼠96隻,隨機(隨機數字法)分為3組:假手術組(Sham組,n=32)、膿毒癥組(CLP組,n=32)、EPO組(n=32,膿毒癥模型基礎上EPO 1000 IU/kg腹腔內註射).分彆于造模術後3、6、12、24 h記錄心髒血流動力學指標變化,測定血清炎癥反應因子水平及心肌酶變化,應用流式細胞儀檢測心肌細胞線粒體膜電位、心肌細胞凋亡及心肌組織覈因子-κB (NF-κB p65)錶達水平；光鏡下觀察大鼠心肌細胞的病理學變化.所有數據以均數±標準差(-x±s)錶示,採用SPSS 13.0統計軟件分析,組間比較採用t檢驗,以P ＜0.05為差異具有統計學意義.結果 (1)與Sham組相比,CLP組大鼠左室收縮壓(LVSP)、左室舒張末壓(LVEDP)、左室壓力上升最大速度(+dp/dtmax)、左室壓力下降最大速度(-dp/dtmax)均有不同程度噁化(P ＜0.05,P＜0.01),EPO組各指標降低幅度小于CLP組(P＜0.05,P＜0.01)；(2)與Sham組相比,CLP組各時間點血清腫瘤壞死因子-α(TNF-α)、白介素-6 (IL-6)、C-反應蛋白水平(CRP)、肌鈣蛋白I(cTNI)、肌痠激酶(CK)、乳痠脫氫酶(LDH)和穀草轉氨酶(AST)水平及心肌組織NF-κB p65錶達水平等指標均有不同程度升高(P＜0.05),而EPO組上述各指標低于CLP組(P＜0.05),與之相反,EPO組血清抗炎因子白介素-10 (IL-10)各時間點均高于CLP組；(3)與Sham組相比,CLP組心肌細胞凋亡率升高(P＜0.01)；(4)心肌細胞線粒體膜電位較Sham組降低(P＜0.01),與CLP組相比,EPO組心肌細胞凋亡率明顯降低,線粒體膜電位升高(P＜0.叭)；(5)應用EPO榦預後,心髒組織損害的病理改變較膿毒癥組減輕.結論 膿毒癥大鼠中,外源性EPO可阻抑心肌細胞線粒體膜電位降低,抑製心肌細胞凋亡；同時抑製NF-κB活化,減少促炎因子生成,減輕心髒組織的炎癥損傷.以上兩種機製髮揮保護受損心肌、改善心功能的作用.
목적 탐토촉홍세포생성소(EPO)대농독증대서심기손상적보호작용급기궤제.방법 통과맹장결찰천공인류술(CLP)건립농독증대서모형.건강SD대서96지,수궤(수궤수자법)분위3조:가수술조(Sham조,n=32)、농독증조(CLP조,n=32)、EPO조(n=32,농독증모형기출상EPO 1000 IU/kg복강내주사).분별우조모술후3、6、12、24 h기록심장혈류동역학지표변화,측정혈청염증반응인자수평급심기매변화,응용류식세포의검측심기세포선립체막전위、심기세포조망급심기조직핵인자-κB (NF-κB p65)표체수평；광경하관찰대서심기세포적병이학변화.소유수거이균수±표준차(-x±s)표시,채용SPSS 13.0통계연건분석,조간비교채용t검험,이P ＜0.05위차이구유통계학의의.결과 (1)여Sham조상비,CLP조대서좌실수축압(LVSP)、좌실서장말압(LVEDP)、좌실압력상승최대속도(+dp/dtmax)、좌실압력하강최대속도(-dp/dtmax)균유불동정도악화(P ＜0.05,P＜0.01),EPO조각지표강저폭도소우CLP조(P＜0.05,P＜0.01)；(2)여Sham조상비,CLP조각시간점혈청종류배사인자-α(TNF-α)、백개소-6 (IL-6)、C-반응단백수평(CRP)、기개단백I(cTNI)、기산격매(CK)、유산탈경매(LDH)화곡초전안매(AST)수평급심기조직NF-κB p65표체수평등지표균유불동정도승고(P＜0.05),이EPO조상술각지표저우CLP조(P＜0.05),여지상반,EPO조혈청항염인자백개소-10 (IL-10)각시간점균고우CLP조；(3)여Sham조상비,CLP조심기세포조망솔승고(P＜0.01)；(4)심기세포선립체막전위교Sham조강저(P＜0.01),여CLP조상비,EPO조심기세포조망솔명현강저,선립체막전위승고(P＜0.팔)；(5)응용EPO간예후,심장조직손해적병리개변교농독증조감경.결론 농독증대서중,외원성EPO가조억심기세포선립체막전위강저,억제심기세포조망；동시억제NF-κB활화,감소촉염인자생성,감경심장조직적염증손상.이상량충궤제발휘보호수손심기、개선심공능적작용.
Objective To investigate the protective effects of erythropoietin (EPO) on myocardium injury after sepsis in rats in order to clarify the mechanisms.Methods The rat models of sepsis were produced by cecal ligation and perforation method (CLP).Ninty-six healthy SD rats were randomly (random number) divided into 3 groups:the sham operation group (Sham group,n =32),the sepsis group (CLP group,n =32),and the EPO treatment group (EPO group,n =32) treated with EPO 1000 IU/kg intraperitoneal injection after the CLP.The observation intervals were set at 3 h,6 h,12 h and 24 h after the surgery.The cardiac hemodynamics of the CLP group were measured.Plasma levels of inflammatory factors and myocardial enzyme indicators were determined by enzyme-linked immunosorbent assay (ELISA) ; Membrane potential levels of chondriosome of myocardial cells,cell apoptosis rates and expressions of nuclear factor-κB p65 of myocardium tissue were detected by flow cytometer; Then the pathological change of myocardium with HE staining was observed under light microscopy.Results (1) Compared with the CLP group,left ventricle systolic pressure (LVSP),left ventricle diastolic end pressure (LVEDP),the maximum rate of left ventricle rise and fall (+ dp/dtmax and-dp/dtmax) in the EPO group improved (P ＜0.05,P＜0.01); (2) Compared with the Sham operation group,plasma levels of tumor necrosis factoralpha (TNF-α),interleukin-6 (IL-6),C-reactive protein (CRP),cardiac troponin-I (cTNI),creatine kinase (CK),lactate dehydrogenase (LDH) and glutamine-oxaloacetic transaminase (GOT) in the CLP group increased at each interval (P ＜ 0.05),and those biomarkers in the EPO group were lower than those in the CLP group (P ＜ 0.05) ; On the contrary,plasma level of anti-inflammatory factor IL-10 in EPO group was higher than that in the CLP group (P ＜ 0.01) ; (3) Compared with the Sham operation group,the cell apoptosis rates in the CLP group increased significantly (P ＜ 0.01),and it decreased obviously in the EPO group (P ＜ 0.01); (4) Compared with the Sham group,membrane potential levels of chondriosome of myocardial cell in the CLP group decreased (P ＜0.01),while it increased in the EPO group in comparison with the CLP group (P ＜ 0.01) ; (5) Pathological changes of myocardium after the CLP could be lessened by the EPO treatment.Conclusions EPO may increase membrane potential levels of chondriosome and decrease the apoptosis rates of myocardial cell in rats after sepsis,and it may reduced the production of inflammatory factors to protect the myocardial cell by down-regulating NF-κB p65.Both increased membrane potential level of chonriosome and decreased inflammatory factor may implicate in myocardium protection thereby improving cardiac function after sepsis.
