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学科主题临床检验诊断学
自噬对大鼠肾缺血再灌注损伤的保护作用及机制
其他题名Protective role of autophagy in rat renal ischemia-reperfusion injury and its related mecha-nisms
张雅丽; 崔丽艳; 杨硕; 周剑锁; 张文静; 张捷
关键词自噬 缺血再灌注 凋亡 免疫组织化学 超微结构 autophagy ischemia-reperfusion apoptosis immunohistochemisty ultrastructure
刊名临床与实验病理学杂志
2015
DOI10.13315/j.cnki.cjcep.2015.04.017
4页:431-435
收录类别中国科技核心期刊 ; 中文核心期刊 ; CSCD
文章类型Journal Article
摘要目的:观察大鼠肾脏缺血再灌注( ischemia reperfusion, I/R)损伤中肾小管上皮细胞的自噬激活情况,探讨其对肾脏I/R损伤发挥保护性作用的分子机制。方法将40只雄性Wistar大鼠随机分为4组:假手术组( Sham)、I/R组、氯喹干预组( I/R+CQ)和雷帕霉素干预组( I/R+Rap)。常规方法建立大鼠肾脏I/R损伤模型,再灌注24 h后留取各组大鼠血和肾脏标本。血标本用于尿素氮( BUN)和血肌酐( Scre)含量检测;肾组织标本行HE染色,观察病理学损伤情况;TUNEL法检测肾小管上皮细胞凋亡的改变;采用免疫组化法检测Beclin-1和Caspase-3蛋白表达,透射电镜观察大鼠肾脏自噬泡的形成情况。结果与I/R组相比,I/R+CQ组BUN和Scre明显升高(P<0.05,P<0.01),肾组织损伤积分增加(P<0.01),TUNEL阳性细胞数增多(P<0.05),Beclin-1蛋白表达减弱(P<0.01),Caspase-3蛋白表达增强(P<0.01),电镜下自噬泡数量较少(P<0.05);I/R+Rap组BUN和Scre显著降低(P<0.01),肾组织损伤积分减低(P<0.05),TUNEL阳性细胞数减少(P<0.05),Beclin-1蛋白表达增强(P<0.01),Caspase-3蛋白表达减弱(P<0.01),自噬泡数量明显增多(P<0.01)。结论自噬激活在肾脏I/R损伤过程中可通过抑制凋亡而发挥的保护作用,其机制可能涉及Beclin-1及Caspase-3等蛋白的表达调控。 Purpose To observe the autophagy activation of renal tubular epithelial cells during rat renal ischemia-reperfusion ( I/R) injury, and then explore the possible mechanisms of the protective role of autophagy. Methods Forty male Wistar rats were randomly divided into the following four groups: (1) sham operation group (Sham), (2) I/R, (3) chloroquine intervention group (I/R+CQ), (4) rapamycin intervention group (I/R +Rap). The specimens of blood and kidney were collected after reperfusion of 24 hours. The serum creatinine ( Scre) and blood urea nitrogen ( BUN) levels were measured from blood samples. Tissue samples of the kidney were stained with HE to observe the pathological changes. Apoptotic cells in the kidney sections were detected using the termi-nal deoxynucleotidyl transferase dUTP nick end labeling ( TUNEL) assay. Immunohistochemistry was used to detect the expression of Beclin-1 and Caspase-3. The structures of autophagic vacuoles were revealed by transmission electron microscopy. Results Compared to I/R group, the I/R+CQ group presented higher levels of BUN and Scre (P<0. 05, P<0. 01), higher renal tissue injury scores (P<0. 01), increased TUNEL-positive cells (P<0. 05), down-regulation of Beclin-1 (P<0. 01) and up-regulation of Caspase-3 (P<0. 01), as well as reduced number of autophagic vacuoles (P<0. 05). On the contrary, I/R+Rap group exhibited lower levels of BUN and Scre (P<0. 01), decreased renal tissue injury scores (P<0. 05), increased TUNEL-positive cells (P<0. 05), up-regula-tion of Beclin-1 (P<0. 01) and down-regulation of Caspase-3 (P<0. 01), as well as increased autophagic vacuoles (P<0. 05). Conclusion Activated autophagy provided a protective role in the kidney I/R injury through inhibiting apoptosis, and the mechanisms may be involved in the regulation of Beclin-1 and Caspase-3.
语种中文
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文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/71642
专题北京大学第三临床医学院_检验科
作者单位北京大学第三医院检验科,北京,100191
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张雅丽,崔丽艳,杨硕,等. 自噬对大鼠肾缺血再灌注损伤的保护作用及机制[J]. 临床与实验病理学杂志,2015(4):431-435.
APA 张雅丽,崔丽艳,杨硕,周剑锁,张文静,&张捷.(2015).自噬对大鼠肾缺血再灌注损伤的保护作用及机制.临床与实验病理学杂志(4),431-435.
MLA 张雅丽,et al."自噬对大鼠肾缺血再灌注损伤的保护作用及机制".临床与实验病理学杂志 .4(2015):431-435.
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