吉林大学学报(医学版)

• 基础研究 • 上一篇    下一篇

七叶皂苷钠对肠缺血再灌注损伤大鼠p38MAPK信号通路的影响及其保护作用机制

王艳蕾,吴丽,李筠璐,秦丽娟,李树民,孙娜   

  1. 河北联合大学基础医学院生理学系,河北 唐山063000  
  • 收稿日期:2013-05-19 出版日期:2014-01-28 发布日期:2014-01-25
  • 通讯作者: 王艳蕾(Tel:0315-3725606,E-mail:wangyanlei2013@126.com) E-mail:wangyanlei2013@126.com
  • 作者简介:王艳蕾(1973-),女,河北省唐山市人,副教授,医学硕士,主要从事缺血再灌注损伤机制及治疗的研究。
  • 基金资助:

    河北省科技厅科研基金资助课题(112761172);河北省唐山市科学技术研究与发展计划项目资助课题(09130202A-3-7)

Influence of sodium aescinate in p38MAPK pathway in rats with intestinal ischemia reperfusion injury and mechanism of its protective effect

WANG Yan-lei,WU Li,LI Jun-lu,QIN Li-juan,LI Shu-min,SUN Na   

  1. Department of Physiology,School of Basic Medical Sciences,Hebei United University,Tangshan 063000,China
  • Received:2013-05-19 Online:2014-01-28 Published:2014-01-25

摘要:

目的:观察七叶皂苷钠(SA)对肠缺血再灌注(I/R)损伤大鼠p38MAPK信号通路的影响,阐明其对肠I/R损伤保护作用的机制。方法:采用夹闭肠系膜上动脉(SMA)方法建立大鼠肠I/R损伤模型。24只SD大鼠随机分为对照组(只分离出SMA,不行夹闭)、I/R组(分离出SMA,在其根部用无创伤血管夹夹闭阻断该动脉血运1 h后松夹,实现再灌注)和SA组(手术过程同I/R组)。分别于夹闭前、松夹前、松夹后30 min 3个时间点对各组每只大鼠给予如下处理,对照组和I/R组大鼠尾静脉推注生理盐水5 mL/kg;SA组大鼠尾静脉推注SA 0.9 mg/Kg。再灌注后1 h测定各组大鼠血浆肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平以及血浆和小肠组织中髓过氧化物酶(MPO)、二胺氧化酶(DAO)活性和丙二醛(MDA)水平;同时免疫组织化学法检测各组大鼠小肠组织中p38MAPK和Bax蛋白的表达水平。结果:与对照组比较,I/R组大鼠血浆MPO、DAO活性及MDA、TNF-α、IL-6水平升高,小肠组织MPO活性及MDA水平升高,DAO活性降低(P<0.01);与I/R组比较,SA组大鼠血浆MPO、DAO的活性以及MDA、TNF-α和IL-6水平降低,小肠组织MPO活性及MDA水平降低,DAO活性升高(P<0.01)。免疫组织化学检测,与对照组比较,I/R组和SA组大鼠小肠组织p38MAPK和Bax的蛋白表达明显增强(P<0.01);与I/R组比较,SA组大鼠小肠组织p38MAPK和Bax的蛋白表达下降(P<0.01)。相关分析显示,大鼠小肠组织MPO、MDA、Bax蛋白表达水平以及血浆TNF-α、IL-6水平与肠组织细胞p38MAPK蛋白表达水平呈正相关关系(r分别为0.797、0.702、0.712、0.695和0.715,P<0.01)。结论:SA可能通过抑制氧自由基、炎性细胞因子的产生以及中性粒细胞的活化,下调肠组织p38MAPK的表达,从而抑制肠组织细胞凋亡,减轻肠I/R时的肠损伤。

关键词: 七叶皂苷钠, 小肠, 缺血再灌注, p38MAPK, 细胞凋亡, 细胞因子

Abstract:

Abstract:Objective  To observe the effect of sodium aescinate(SA) on p38MAPK pathway in rats with intestinal ischemia reperfusion(I/R) injury, and to charify the protective mechanism of SA. Methods Twenty-four SD rats were randomly divided into three groups with eight rats in each:control group,intestinal I/R group,SA group.In control group,superior mesenteric artery (SAM)  was separated but not clamped;in I/R and SA groups,SMA was separated and clamped for 60 min.Each rat was given different treatment before ischemia,before reperfusion and 30 min after reperfusion respectively.In control and I/R groups,saline(5 mL/kg) was injected through tail;in SA group,SA(0.9 mg/kg) was injected through tail. The levels of TNF-α,IL-6 in plasma and the activities of myeloperoxidase(MPO) and diamine oxidase(DAO),malondialdehyde(MDA) levels in plasma and intestinal tissue were measured 60 min after reperfusion,as well as the expression levels of p38MAPK and Bax proteins in intestinal tissue were examined using immunohistochemical method.Results Compared with control group,the activities of MPO,DAO and the levels of TNF-α,IL-6,MDA in plasma were significantly increased;and the activity of MPO and the  MDA levels in intestinal tissue were also significantly increased,and while the DAO activity in intestinal tissue was significantly decreased in I/R group(all P<0.01).Compared with I/R group,the activities of MPO and DAO and the levels of MDA,TNF-α,IL-6 in plasma were significantly decreased;the MPO  activity and MDA level in intestinal tissue were also significantly decreased,and while the DAO activity in intestinal tissue was significantly increased in SA group(P<0.01).The immunohistochemistry results indicated that compared with control group,the protein expression levels of p38MAPK and Bax were significantly increased in I/R and SA groups(P<0.01);compared with I/R group,the p38MAPK and Bax protein expression levels were significantly decreased in SA group(P<0.01).The correlation analysis indicated that the MPO activity,the MDA level  and the expression levels of Bax protein in intestinal tissue  and the TNF-α,IL-6 levels  in plasma were positively correlated with the expression level of p38MAPK protein (r=0.797,0.702,0.712,0.695 and 0.715,all P<0.01).Conclusion SA can protect the intestinal I/R injury,which may be mediated by reducing the release of oxygen free radicals and cytokine,inhibiting activation of neutrophils,down-regulating p38MAPK gene protein expression to inhibit intestinal tissue apoptosis.

Key words:  , sodium aescinate, intestine, ischemia/reperfusion, p38MAPK, apoptosis, cytokine

中图分类号: 

  • R656.7