CCN1对小鼠肝脏缺血再灌注损伤的影响
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国家自然科学基金(81270944)


Matricellular protein CCN1 aggravates hepatic ischemia-reperfusion injury in mice
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    摘要:

    目的:研究细胞外基质蛋白CCN1(cysteine-rich protein 61,Cyr61)在小鼠的缺血再灌注(ischemia-reperfusion,IR)损伤中的表达变化和作用。方法:建立小鼠70%肝脏IR模型。肝脏缺血60 min后再灌注,于不同时间点通过qRT-PCR和Western blot检测CCN1的mRNA和蛋白表达水平。部分实验中,在IR前给予小鼠尾静脉注射CCN1重组蛋白或生理盐水,通过转氨酶和病理学检测,比较两组小鼠肝脏IR损伤情况;收集肝脏组织,用qRT-PCR检测炎症因子的表达水平。结果:与sham组相比,肝缺血60 min再灌注3-6-18 h后CCN1的表达明显升高;注射CCN1蛋白的小鼠肝IR 6 h后,与生理盐水组相比,血清丙氨酸氨基转移酶-天门冬氨酸氨基转移酶水平升高,病理改变更严重。注射CCN1小鼠的肝组织中白介素-6-CXCL-10的表达水平也显著升高。结论:CCN1在小鼠肝IR损伤中表达显著增加,并可能通过诱导炎症因子的表达对IR损伤起促进作用。

    Abstract:

    Objective:To investigate the expression and function of CCN1 in hepatic ischemia-reperfusion (I/R)injury in mice. Methods:Partial warm ischemia was produced in the left and middle hepatic lobes of mice for 60 min,followed by reperfusion. Expression of CCN1 was detected by real-time PCR and Western blot at timed points. In indicated experiments,recombinant CCN1 or saline was injected via tail vein 1 h before hepatic ischemia. Levels of alanine aminotransferase (ALT)and aspartate aminotransferase(AST)in blood were examined at 6 h of reperfusion. Pathological analysis was performed with liver sections after hematoxylin and eosin staining. mRNA levels of cytokines were detected by real-time PCR. Results:CCN1 expression is up-regulated in hepatic I/R. Serum ALT and AST levels were significantly increased after CCN1 treatment. The pathological alterations were more severe compared with saline group. Gene transcripts for proinflammatory cytokines in liver tissue were elevated significantly in the CCN1 group. Conclusion:Induction of CCN1 in liver I/R aggravates hepatic injury,possibly by promoting the expression of inflammatory factors.

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林燕蝶,孙 武,李 皓. CCN1对小鼠肝脏缺血再灌注损伤的影响[J].南京医科大学学报(自然科学版),2016,(7):806-810

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  • 收稿日期:2016-03-03
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  • 在线发布日期: 2016-07-15
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