中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2012年
7期
430-434
,共5页
李奎%高健刚%祝海%贾勇%孙延波%侯四川
李奎%高健剛%祝海%賈勇%孫延波%侯四川
리규%고건강%축해%가용%손연파%후사천
左卡尼汀%肾%缺血再灌注损伤%NF-E2相关因子2
左卡尼汀%腎%缺血再灌註損傷%NF-E2相關因子2
좌잡니정%신%결혈재관주손상%NF-E2상관인자2
Levocarnidine%Kidney%Ischemia reperfusion injury%NF-E2-related factor 2
目的 研究左卡尼汀对大鼠肾缺血再灌注损伤(IRI)的影响及其机制.方法 将Wistar大鼠分为3组:L组大鼠制成IRI模型,于夹闭肾动静脉前5 min及松开动脉夹后30 min,分2次经尾静脉注射左卡尼汀,各500mg/kg;I组大鼠制成IRI模型,仅注射生理盐水；C组仅分离双侧肾动、静脉,注射生理盐水.分别于再灌注后3、6和24 h处死各组大鼠.处死前经下腔静脉取血,检测血清肌酐(Cr)、尿素氮( BUN)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量.获取肾组织样本,进行病理学观察；应用逆转录聚合酶链反应检测肾组织核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、γ-谷氨酰半胱氨酸合成酶(γ-GCS)的mRNA水平；蛋白质印迹法检测细胞核中Nrf2含量；免疫组织化学法检测肾组织中Nrf2的表达及定位.结果 再灌注后3h时,L组与I组血清Cr和BUN均高于C组(P＜0.01)；再灌注后6h时,L组血清Cr和BUN高于C组(P＜0.01),而低于I组(P＜0.01)；再灌注后24 h时,L组血清Cr和BUN仍低于I组(P＜0.05).再灌注后6和24 h时,L组与I组SOD水平低于C组(P＜0.05),MDA水平高于C组(P＜0.05).再灌注后各时间点,L组SOD水平均高于I组(P＜0.05),MDA水平均低于I组(P＜0.05).再灌注后24 h时,L组肾组织病理改变较I组轻.再灌注后6h时,I组Nrf2、HO-1、γ-GCS的mRNA相对含量均高于C组(P＜0.05),而L组各基因mRNA的相对含量高于I组(P＜0.05).L组细胞核内Nrf2的相对含量高于I组(P＜0.05).结论 左卡尼汀可减轻大鼠肾脏IRI,其机制可能与激活Nrf2-ARE通路,进而增强下游抗氧化基因的表达有关.
目的 研究左卡尼汀對大鼠腎缺血再灌註損傷(IRI)的影響及其機製.方法 將Wistar大鼠分為3組:L組大鼠製成IRI模型,于夾閉腎動靜脈前5 min及鬆開動脈夾後30 min,分2次經尾靜脈註射左卡尼汀,各500mg/kg;I組大鼠製成IRI模型,僅註射生理鹽水；C組僅分離雙側腎動、靜脈,註射生理鹽水.分彆于再灌註後3、6和24 h處死各組大鼠.處死前經下腔靜脈取血,檢測血清肌酐(Cr)、尿素氮( BUN)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量.穫取腎組織樣本,進行病理學觀察；應用逆轉錄聚閤酶鏈反應檢測腎組織覈因子E2相關因子2(Nrf2)、血紅素加氧酶-1(HO-1)、γ-穀氨酰半胱氨痠閤成酶(γ-GCS)的mRNA水平；蛋白質印跡法檢測細胞覈中Nrf2含量；免疫組織化學法檢測腎組織中Nrf2的錶達及定位.結果 再灌註後3h時,L組與I組血清Cr和BUN均高于C組(P＜0.01)；再灌註後6h時,L組血清Cr和BUN高于C組(P＜0.01),而低于I組(P＜0.01)；再灌註後24 h時,L組血清Cr和BUN仍低于I組(P＜0.05).再灌註後6和24 h時,L組與I組SOD水平低于C組(P＜0.05),MDA水平高于C組(P＜0.05).再灌註後各時間點,L組SOD水平均高于I組(P＜0.05),MDA水平均低于I組(P＜0.05).再灌註後24 h時,L組腎組織病理改變較I組輕.再灌註後6h時,I組Nrf2、HO-1、γ-GCS的mRNA相對含量均高于C組(P＜0.05),而L組各基因mRNA的相對含量高于I組(P＜0.05).L組細胞覈內Nrf2的相對含量高于I組(P＜0.05).結論 左卡尼汀可減輕大鼠腎髒IRI,其機製可能與激活Nrf2-ARE通路,進而增彊下遊抗氧化基因的錶達有關.
목적 연구좌잡니정대대서신결혈재관주손상(IRI)적영향급기궤제.방법 장Wistar대서분위3조:L조대서제성IRI모형,우협폐신동정맥전5 min급송개동맥협후30 min,분2차경미정맥주사좌잡니정,각500mg/kg;I조대서제성IRI모형,부주사생리염수；C조부분리쌍측신동、정맥,주사생리염수.분별우재관주후3、6화24 h처사각조대서.처사전경하강정맥취혈,검측혈청기항(Cr)、뇨소담( BUN)、초양화물기화매(SOD)급병이철(MDA)함량.획취신조직양본,진행병이학관찰；응용역전록취합매련반응검측신조직핵인자E2상관인자2(Nrf2)、혈홍소가양매-1(HO-1)、γ-곡안선반광안산합성매(γ-GCS)적mRNA수평；단백질인적법검측세포핵중Nrf2함량；면역조직화학법검측신조직중Nrf2적표체급정위.결과 재관주후3h시,L조여I조혈청Cr화BUN균고우C조(P＜0.01)；재관주후6h시,L조혈청Cr화BUN고우C조(P＜0.01),이저우I조(P＜0.01)；재관주후24 h시,L조혈청Cr화BUN잉저우I조(P＜0.05).재관주후6화24 h시,L조여I조SOD수평저우C조(P＜0.05),MDA수평고우C조(P＜0.05).재관주후각시간점,L조SOD수평균고우I조(P＜0.05),MDA수평균저우I조(P＜0.05).재관주후24 h시,L조신조직병리개변교I조경.재관주후6h시,I조Nrf2、HO-1、γ-GCS적mRNA상대함량균고우C조(P＜0.05),이L조각기인mRNA적상대함량고우I조(P＜0.05).L조세포핵내Nrf2적상대함량고우I조(P＜0.05).결론 좌잡니정가감경대서신장IRI,기궤제가능여격활Nrf2-ARE통로,진이증강하유항양화기인적표체유관.
Objective To investigate the effect of L-camitine on renal ischemia-reperfusion (IR) injury (IRI) and Nrf2-ARE signaling pathway in rats.Methods Rats were randomly separated into the following experimental groups:control group (group C),IRI group (group I) and L-carnitine group (group L).Rats accepted no treatment of ischemic reperfusion in group C.In groups I and group L,the renal IRI model was established.L-carnitine was injected through the tail vein in group L,while the equal volume of saline was injected in group C and group I.Rats were killed at 3,6,and 24 h after IR.The levels of serum creatinine (Cr) and blood urea nitrogen (BUN),the activity of superoxide dismutase (SOD) and the content of malonaldehyde (MDA) in serum were measured.The histopathological lesions were observed in renal tissues after 24-h IR.RT-PCR was used to detect the levels of Nrf2,HO-1 and γ-GCS mRNA.Western blotting and immunohistochemistry were used to detect the levels and localization of Nrf2 protein in renal tissues after 6-h IR.Results The levels of Cr and BUN in group I and group L were higher than those in group C at 3 h after IR.At 6 h after IR,the levels of Cr and BUN in group L were lower than those in group I (P＜0.01 ).At 24 h after IR,the levels of Cr and BUN in group L were still lower than those in group I though both of them were reduced (P＜0.05).At all time points,the activity of SOD in group L was higher and the content of MDA was lower than those in group I (P＜ 0.05). As compared with group I,the renal histopathological lesions were alleviated in group L at 24 h after IR.At 6 h after IR,levels of Nrf2,HO-1,γ-GCS mRNA and Nrf2 protein in group I were increased as compared with group C,but decreased as compared with group L.Beyond that,the expression of nuclear Nrf2 protein in group L was higher than that in group I.Conclusion L-carnitine can protects the kidney against IRI significantly,which may be due to the up-regulated expression of antioxidant genes by activating the Nrf2-ARE signaling pathway.
