中华胸心血管外科杂志
中華胸心血管外科雜誌
중화흉심혈관외과잡지
Chinese Journal of Thoracic and Cardiovascular Surgery
2014年
4期
234-237
,共4页
李占清%伊雪%邬鹏宇%王帅%杨学慧%崔翔宇%刘志勇%郑素琴%杨方
李佔清%伊雪%鄔鵬宇%王帥%楊學慧%崔翔宇%劉誌勇%鄭素琴%楊方
리점청%이설%오붕우%왕수%양학혜%최상우%류지용%정소금%양방
大鼠%心脏移植%心肌再灌注损伤%细胞凋亡%莱菔硫烷
大鼠%心髒移植%心肌再灌註損傷%細胞凋亡%萊菔硫烷
대서%심장이식%심기재관주손상%세포조망%래복류완
Rats%Heart transplantation%Reperfusion injury%Apoptosis%Sulforaphane
目的 建立大鼠同种异体异位心脏移植模型,莱菔硫烷(Sulforaphane,SFN)预处理受体大鼠,观察并探讨其对大鼠心肌冷缺血再灌注损伤的保护作用.方法 健康雄性Lewis大鼠140只,其中100只采用数字表法随机分为对照组、SFN0.5组、SFN0.75组、SFN1.0组和SFN25组5组,每组20只(供、受体各10只).各组受体大鼠分别于受体移植前24h经尾静脉注射等量生理盐水、SFN0.5mg· kg-1 bw-1、SFN 0.75mg·kg-·bw-、SFN 1.0mg·kg-1 ·bw-和SFN 2.5 mg·kg-1·bw-.将冷藏于4C HTK液18h的供心,移植到受体大鼠腹腔内,建立大鼠同种异体异位心脏移植模型.移植后6、24 h分别从受体鼠眼内眦静脉或腔静脉采血,检测血清中乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)、肌钙蛋白(TnT)水平;移植后24 h,取供心心肌组织,检测心肌组织中脂质过氧化物(lipid hydroperoxide,LPO)含量及超氧化物歧化酶(superoxide dismutase,SOD)的活性,应用免疫组织化学检测移植心肌组织诱导型一氧化氮合酶(inducible Nitric Oxide,iNOS)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、低氧诱导因子-1oα(Hypoxia in-ducible factor,HIF-1α)的表达,电镜观察心肌超微结构变化.另外40只大鼠应用Kaplan-meier分析其生存率.结果 与对照组大鼠比较,其他4组经SFN预处理大鼠的中位生存期明显高;血清中LDH、TnT、CK-MB水平均有不同程度下降;心肌LPO含量均降低(P＜0.05);SOD/LPO比值高(P＜0.01);心肌超微结构损伤程度轻.另外,与对照组比较,SFN0.75组、SFN10组(P＜0.05)、SFN2.5组(P＜0.05)大鼠的心脏功能评分较高;移植后6、24 h,SFN0.5组心肌组织SOD活性升高(P＜0.05);SFN0.75组、SFN1.0组、SFN2.5组心肌组织iNOS表达降低(P＜0.01);SFN1.0组、SFN2.5组Caspase-3表达低(P＜0.05);SFN10组、SFN25组HIF-1α表达低(P＜0.05).结论 SFN可通过抗氧化、抗细胞凋亡机制,减轻心脏移植中冷缺血再灌注损伤引起的氧化应激和心肌细胞凋亡,从而减轻移植心脏的缺血再灌注损伤.SFN剂量增加,保护作用增强.
目的 建立大鼠同種異體異位心髒移植模型,萊菔硫烷(Sulforaphane,SFN)預處理受體大鼠,觀察併探討其對大鼠心肌冷缺血再灌註損傷的保護作用.方法 健康雄性Lewis大鼠140隻,其中100隻採用數字錶法隨機分為對照組、SFN0.5組、SFN0.75組、SFN1.0組和SFN25組5組,每組20隻(供、受體各10隻).各組受體大鼠分彆于受體移植前24h經尾靜脈註射等量生理鹽水、SFN0.5mg· kg-1 bw-1、SFN 0.75mg·kg-·bw-、SFN 1.0mg·kg-1 ·bw-和SFN 2.5 mg·kg-1·bw-.將冷藏于4C HTK液18h的供心,移植到受體大鼠腹腔內,建立大鼠同種異體異位心髒移植模型.移植後6、24 h分彆從受體鼠眼內眥靜脈或腔靜脈採血,檢測血清中乳痠脫氫酶(LDH)、肌痠激酶同工酶(CK-MB)、肌鈣蛋白(TnT)水平;移植後24 h,取供心心肌組織,檢測心肌組織中脂質過氧化物(lipid hydroperoxide,LPO)含量及超氧化物歧化酶(superoxide dismutase,SOD)的活性,應用免疫組織化學檢測移植心肌組織誘導型一氧化氮閤酶(inducible Nitric Oxide,iNOS)、半胱氨痠天鼕氨痠蛋白酶3(Caspase-3)、低氧誘導因子-1oα(Hypoxia in-ducible factor,HIF-1α)的錶達,電鏡觀察心肌超微結構變化.另外40隻大鼠應用Kaplan-meier分析其生存率.結果 與對照組大鼠比較,其他4組經SFN預處理大鼠的中位生存期明顯高;血清中LDH、TnT、CK-MB水平均有不同程度下降;心肌LPO含量均降低(P＜0.05);SOD/LPO比值高(P＜0.01);心肌超微結構損傷程度輕.另外,與對照組比較,SFN0.75組、SFN10組(P＜0.05)、SFN2.5組(P＜0.05)大鼠的心髒功能評分較高;移植後6、24 h,SFN0.5組心肌組織SOD活性升高(P＜0.05);SFN0.75組、SFN1.0組、SFN2.5組心肌組織iNOS錶達降低(P＜0.01);SFN1.0組、SFN2.5組Caspase-3錶達低(P＜0.05);SFN10組、SFN25組HIF-1α錶達低(P＜0.05).結論 SFN可通過抗氧化、抗細胞凋亡機製,減輕心髒移植中冷缺血再灌註損傷引起的氧化應激和心肌細胞凋亡,從而減輕移植心髒的缺血再灌註損傷.SFN劑量增加,保護作用增彊.
목적 건립대서동충이체이위심장이식모형,래복류완(Sulforaphane,SFN)예처리수체대서,관찰병탐토기대대서심기랭결혈재관주손상적보호작용.방법 건강웅성Lewis대서140지,기중100지채용수자표법수궤분위대조조、SFN0.5조、SFN0.75조、SFN1.0조화SFN25조5조,매조20지(공、수체각10지).각조수체대서분별우수체이식전24h경미정맥주사등량생리염수、SFN0.5mg· kg-1 bw-1、SFN 0.75mg·kg-·bw-、SFN 1.0mg·kg-1 ·bw-화SFN 2.5 mg·kg-1·bw-.장랭장우4C HTK액18h적공심,이식도수체대서복강내,건립대서동충이체이위심장이식모형.이식후6、24 h분별종수체서안내자정맥혹강정맥채혈,검측혈청중유산탈경매(LDH)、기산격매동공매(CK-MB)、기개단백(TnT)수평;이식후24 h,취공심심기조직,검측심기조직중지질과양화물(lipid hydroperoxide,LPO)함량급초양화물기화매(superoxide dismutase,SOD)적활성,응용면역조직화학검측이식심기조직유도형일양화담합매(inducible Nitric Oxide,iNOS)、반광안산천동안산단백매3(Caspase-3)、저양유도인자-1oα(Hypoxia in-ducible factor,HIF-1α)적표체,전경관찰심기초미결구변화.령외40지대서응용Kaplan-meier분석기생존솔.결과 여대조조대서비교,기타4조경SFN예처리대서적중위생존기명현고;혈청중LDH、TnT、CK-MB수평균유불동정도하강;심기LPO함량균강저(P＜0.05);SOD/LPO비치고(P＜0.01);심기초미결구손상정도경.령외,여대조조비교,SFN0.75조、SFN10조(P＜0.05)、SFN2.5조(P＜0.05)대서적심장공능평분교고;이식후6、24 h,SFN0.5조심기조직SOD활성승고(P＜0.05);SFN0.75조、SFN1.0조、SFN2.5조심기조직iNOS표체강저(P＜0.01);SFN1.0조、SFN2.5조Caspase-3표체저(P＜0.05);SFN10조、SFN25조HIF-1α표체저(P＜0.05).결론 SFN가통과항양화、항세포조망궤제,감경심장이식중랭결혈재관주손상인기적양화응격화심기세포조망,종이감경이식심장적결혈재관주손상.SFN제량증가,보호작용증강.
Objective To explore the protection of Sulforaphane on myocardial cold ischemia reperfusion injury in the isogeneicheterotopic heart transplantation model of rats.Methods 100 healthy male Lewis rats were randomly divided into 5groups,20 rats in each group(donor and recipient 10 rats respectively).Control group:recipient rats were injected the equal amountsaline via tail vein 24 h before heart transplantation.SFN0.5 group:recipient rats were injected sulforaphane 0.5 mg· kg-1 · bw-1 via tail vein 24 h before heart transplantation.SFN0.75 group:recipient rats were injected sulforaphane 0.75 mg · kg-1 · bw-1 via tail vein 24 h before heart transplantation.SFN1.0 group:recipient rats were injected sulforaphane 1.0 mg · kg-1 · bw-1 via tail vein 24 h before heart transplantation.SFN2.5 group:recipient rats were injected sulforaphane 2.5 mg · kg-1 · bw-1 via tail vein 24 h before heart transplantation.Donnor rats' hearts were stored in 4℃ HTK solution for 18 h in control and experimental groups.Heart transplantation was performed.Lactate dehydrogenase (LDH),Creatine kinaseMB (CK-MB),Troponin T(TnT) were determined at 6 h and 24 h after heart transplanatation.Levels of LPO and activity SOD were tested at 24 h after heart transplanatation.The protein expression of iNOS,Caspase-3,HIF-1α was detected by immunohistochemistry.The changes of myocardial ultrastructural were detected by transmission electron microscope.The survival of two groups was estimated by kaplan-meier analysis using 40 healthy male Lewis rats.Results The median survival time of SFN preconditioning group was obviously higher than that of control group (7d vs.5d),graft function in SFN0.75,SFN1.0,SFN2.5 groups was significantly improved comnpared with control group.The levels of LDH,CK-MB,TnT at 24 h were significantly reduced in SFN preconditioning groups compared with control group.The level of LPO was significantly decreased in SFN preconditioning groups compared with control group,SOD/LPO was significantly increased in SFN preconditioning groups,but the activity of SOD was significant increased in SFN0.5 group.The protein expression of iNOS,Caspase-3,HIF-1 α was significantly reduced in SFN1.0and SFN2.5 groups compared with control group.Myocardial ultrastructure also proved that SFN preconditioning groups was better than control group.Conclusion SFN reduced oxidative stress and myocardial cells apoptosis caused by cold ischemia-reperfusion injury in heart transplantation through its anti-oxidation and anti-apoptotic,which reduced ischemia-reperfusion injury in heart transplantation,and its protective effect increased with dose increased.
