CAJ | 학술논문

目的探讨联合应用来氟米特(LeD和环孢素A(CsA)对大鼠异种心脏移植排斥反应中核因子kB(NF-kB)信号传导通路中核因子出抑制因子激酶α/β(IKKα/β)、NF-kB P65、核因子kB抑制因子α(IkBα)、细胞问粘附分子-1(ICAM-1)表达以及NF-kB DNA结合活性的影响.方法以NIH小鼠为供者,Wismr大鼠为受者,施行颈部心脏移植,CsA组受者术后接受CsA腹腔注射,Lef组受者术后接受Lef灌胃,联合用药组受者术后接受CSA和Lef治疗,另设术后不接受任何药物处理的空白对照组.术后观察移植心脏存活时间,发生排斥反应时,切取移植心脏,进行组织病理学观察,采用免疫组织化学和Western印迹法检测各组移植心肌组织中IKKα/β、NF-kB P65、IkBa和ICAM-1的表达,凝胶电泳迁移率法检测NF-kBDNA结合活性.结果空白对照组、CsA组、Lef组及联合用药组移植心脏的存活时间分别为(2.17±0.41)d、(2.50±1.05)d、(4.17±1.33)d和(6.50±2.56)d,联合用药组明显长于其它3组(P<0.05),Lef组明显长于空白对照组(P<0.05).4个组心肌组织中IKKα/β、NF-KB P65、IKBα和ICAM-1的表达由强到弱依次为空白对照组、CsA组、Lef组、联合用药组,其电泳条带积分吸光度值显示,Lef组的IKKα/β、NF-kB P65、IkBα和ICAM-1的表达明显低于空白对照组和CsA组(P<0.05),而联合用药组明显低于其它3组(P<0.05).Lef组的NF-kBDNA结合活性明显低于空白对照组及CsA组,联合用药组的NF-kDNA结合活性明显低于其它3组,差异均有统计学意义(P<0.05).结论联合应用Lef和CsA可显著延长大鼠移植的异种(小鼠)心脏的存活时间,这可能与NF-kB信号通路中的IKKα/β、NFkB P65、IkBα、ICAM-1表达及NF-kB DNA结合活性受到抑制有关.
목적 탐토연합응용래불미특(LeD화배포소A(CsA)대대서이충심장이식배척반응중핵인자kB(NF-kB)신호전도통로중핵인자출억제인자격매α/β(IKKα/β)、NF-kB P65、핵인자kB억제인자α(IkBα)、세포문점부분자-1(ICAM-1)표체이급NF-kB DNA결합활성적영향.방법 이NIH소서위공자,Wismr대서위수자,시행경부심장이식,CsA조수자술후접수CsA복강주사,Lef조수자술후접수Lef관위,연합용약조수자술후접수CSA화Lef치료,령설술후불접수임하약물처리적공백대조조.술후관찰이식심장존활시간,발생배척반응시,절취이식심장,진행조직병이학관찰,채용면역조직화학화Western인적법검측각조이식심기조직중IKKα/β、NF-kB P65、IkBa화ICAM-1적표체,응효전영천이솔법검측NF-kBDNA결합활성.결과 공백대조조、CsA조、Lef조급연합용약조이식심장적존활시간분별위(2.17±0.41)d、(2.50±1.05)d、(4.17±1.33)d화(6.50±2.56)d,연합용약조명현장우기타3조(P<0.05),Lef조명현장우공백대조조(P<0.05).4개조심기조직중IKKα/β、NF-KB P65、IKBα화ICAM-1적표체유강도약의차위공백대조조、CsA조、Lef조、연합용약조,기전영조대적분흡광도치현시,Lef조적IKKα/β、NF-kB P65、IkBα화ICAM-1적표체명현저우공백대조조화CsA조(P<0.05),이연합용약조명현저우기타3조(P<0.05).Lef조적NF-kBDNA결합활성명현저우공백대조조급CsA조,연합용약조적NF-kDNA결합활성명현저우기타3조,차이균유통계학의의(P<0.05).결론 연합응용Lef화CsA가현저연장대서이식적이충(소서)심장적존활시간,저가능여NF-kB신호통로중적IKKα/β、NFkB P65、IkBα、ICAM-1표체급NF-kB DNA결합활성수도억제유관.
Objective To investigated the effects of leflunomide (Lef) in combination with cyclosporine (CsA) on IKKcomα/β, NF-kB P65, IkBα, ICAM-1 and NF-kB DNA binding activity in NF-kB signal pathway in mouse to rat cardiac xenografts. Methods NIH mice and Wistar rats served as donors and recipients respectively. Mouse to rat heterotopic heart transp lantation was performed (in the neck). The experiments were divided into 4 groups: CsA was injected imraperitoneally after operation in the CsA group; Lef was given in the Lef group by gastric tube after operation; CsA com-bined with Lef was administered after operation in CsA+Lef group; In the control group, the recipi-ents didn't recei veany medicine treatment. After operation, the survival of the transplanted heart was observed. The histophathologic examination was done in the transplanted heart. The expression of IKKα/β, NF-kB P65, IkBα and ICAM-1 and NF-kB DNA binding activity in cardiac xenograft tissues were determined by immunohistochemistry and Western blot as well as electrophoretic mobility shift assay (EMSA) in each group. Results The survival time of the cardiac xenografts in control group,CsA group, Lef group and CsA+Lef group was (2.17±0.41), (2.50±1.05), (4.17±1.33) and (6.50±2.56) days, respectively, significantly longer in CsA + Lef group than in other three groups(P<0.05), and significantly longer in Lef group than in control group (P<0.05). The expression ofIKKα/β (IOD) in cardiac xenograft tissues in control group, CsA group, Lef group and CsA+Lef group was (137.1±4.0), (149.7±6.5), (111.6±14.6) and (81.4±15. 9), the expression ofNF-kB P65 (IOD) was (261.3±31.6), (263.1±28.8), (173.5±5.9) and (67.8+3.8), theexpression of IkBa (IOD) was (88.6±6.7), (80.4±3.5), (64.8±8.9) and (37.3±4.4), theexpression of ICAMol (IOD) was (155.4±5.3), ( 150.7+5.1), ( 104.7±6.2) and (29.2±2.8),and NF-gB DNA binding activity (IOD) was (234.3±8.7), (227.5±14.8), (110.9±12.5) and(43.6±7.4), respectively. The IODs in CsA + Lef group were markedly lower than in the other threegroups (P<0.05), and those in Lef group was obviously lower than in control group (P<0.05).Conclusion The combined use of Lef and CsA can significantly suppress the expression of IKKα/αβ,NF-kB P65, IkBα, ICAM-1 and NF-kB DNA binding activity, which may obviously prolong thesurvival time of mouse to rat cardiac xenografts.