齐墩果酸预处理对大鼠肝脏缺血再灌注损伤过程中IKK/I-κB/NF-κB通路的影响
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国家自然科学基金资助(30672523)


Influence of oleanolic acid pretreatment to the IKK/I-κB/NF-κB signaling pathway during rat’s HIRI
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    摘要:

    目的:探讨齐墩果酸(oleanolic acid,OA)预处理对大鼠肝脏缺血再灌注损伤(hepatic ischemic/reperfusion injury,HIRI)过程中IKK/I-κB/NF-κB信号转导通路的影响-方法:将128只SD大鼠随机分为假手术组(SH组)-缺血再灌注组(IR组)-0.5%羧甲基纤维素钠组(CM组)和齐墩果酸预处理组(OA组)-OA组以100 mg/kg的齐墩果酸混悬液,SH和IR组以相同容积的水,CM组以相同容积的0.5%CMC-Na分别每日灌胃1次,连续7天-第8天建立70%肝脏缺血模型,缺血60 min后再灌注-于术前-再灌注0-3-6 h取肝组织-用Western blot法测定肝脏细胞内IKK2,I-κBα及细胞核内NF-κB p65蛋白含量-结果:术前-0 h,各组胞浆未检测到IKK2,核内仅检测到极少量NF-κB,各组I-κBα蛋白量之间没有统计学差异-3-6 h时,SH组胞浆IKK2和核内NF-κB的蛋白含量均分别明显低于其余3组(P < 0.05);此时OA组该两个蛋白含量分别明显低于CM组和IR组(P < 0.05)-3-6 h时,SH组I-κBα的蛋白含量均分别明显高于其余3组(P < 0.05),OA组此值分别明显高于CM组和IR组(P < 0.05),CM组和IR组各时点之间的细胞内IKK2,I-κBα及细胞核内NF-κB p65蛋白量均无统计学差异(P > 0.05)-结论:HIRI能促进胞浆内IKK2蛋白的表达,使I-κB磷酸化水解,NF-κB活化进入细胞核内介导炎症反应,损伤肝细胞-缺血前使用OA可抑制IKK/I-κB/NF-κB信号传导通路的激活,这可能是OA预处理减轻HIRI的机制之一-

    Abstract:

    Objective:To investigate the influence of oleanolic acid(OA) pretreatment to rat’s IKK/I-κB/NF-κB signaling pathway around its hepatic ischemic/reperfusion injury(HIRI). Methods:One hundred and twenty eight male Sprague-Dawley (SD) rats were randomly divided into sham group(SH),ischemic/reperfusion group(IR),0.5% sodium carboxymethycellulose(CMC-Na) group(CM) and OA+0.5% CMC-Na group(OA). Before the operation,the rats of each group received intragastric administration of corresponding solution once a day for seven days(OA group with 100 mg/kg of OA dissolved in 0.5% CMC-Na,SH group and IR group with water of the same volume,CM group with 0.5% CMC-Na of the same volume). At the 8th day,rats were suffered from segmental(70%) hepatic ischemia for 60 min and then followed with different periods of reperfusion. The hepatic tissue were obtained at 0 h,3 h,and 6 h after the reperfusion. The expression of IKK2 and I-κBα in the cytoplasm and NF-κB p65 in the nucleus were evaluated by the method of Western blotting. Results:There was no IKK2 expression in the cytoplasm,and only few of NF-κB p65 was detected in the nucleus before operation or 0h after reperfusion in each group. Meanwhile,the volume of I-κBα detected in the cytoplasm has no significant difference among four groups at the same point of time. The expression of IKK2 in the cytoplasm and NF-κB p65 in the nucleus in the SH group was much lower than those in other three groups(P < 0.05),while those proteins detected in OA group is less than those in CM group and IR group(P < 0.05)at 3h or 6 h after reperfusion. At these two points of time,the expression of I-κBα in SH group was higher than that in the other three groups(P < 0.05),and this protein detected in OA group is more than that in CM group and IR group(P < 0.05).The expressions of IKK2 and I-κBα in the cytoplasm and NF-κB p65 in nucleus between CM group and IR group had no significant difference at each point of time(P > 0.05). Conclusion:HIRI would increase the expression of IKK2 in the cytoplasm,leading to the phosphorylation of I-κB,activating NF-κB and making it to translocate into the nucleus,and then the activated NF-κB which could mediate inflammatory response eventually caused damage of liver cells. The pretreatment of OA can inhibit the activating of the signaling pathway of IKK/I-κB/NF-κB,which may be one of the mechanisms for OA alleviating HIRI.

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何明枫,华福洲,张 杰,许仄平,孙宏斌,钱燕宁.齐墩果酸预处理对大鼠肝脏缺血再灌注损伤过程中IKK/I-κB/NF-κB通路的影响[J].南京医科大学学报(自然科学版),2011,(5):697-701

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  • 收稿日期:2010-11-30
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