高压氧对大鼠缺血再灌注损伤肾FasL和半胱氨酸蛋白酶3表达的影响  

Effect of hyperbaric oxygen treatment on the expression of FasL and caspase-3 in renaltissue after renal ischemia reperfusion injury

作　　者：孙辉[1] 徐新兵[1] 马凌波[1] 胡光荣[1] 邓颖[1] 王新春[1] 王凤平[1] 

机构地区：[1]哈尔滨医科大学附属第二医院急诊科,150081

出　　处：《中华肾脏病杂志》2012年第10期808-812,共5页Chinese Journal of Nephrology

基　　金：黑龙江省教育厅科研项目（11551194）

摘　　要：目的研究高压氧（HBO）对大鼠缺血再灌注损伤（IRI）肾细胞凋亡相关基因（FasL）和细胞凋亡执行蛋白半胱氨酸蛋白酶3（caspase-3）表达的影响，并探讨其作用机制。方法健康sD雄性大鼠随机分为假手术组（n=8）、IRI组（n=8）和IRI＋HBO组（n=8）。采用夹闭双侧肾动脉方法建立IRI模型。IRI＋HBO组分别在再灌注后1h、24h、48h给予HBO处理，末次HBO后取双肾组织测定各组大鼠肾组织匀浆超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量；采用实时荧光定量PCR和免疫组织化学染色方法分别测定肾组织FasLmRNA、caspase-3蛋白表达。结果与假手术组比较，IRI组SOD活性下降（P〈0．05），MDA含量升高（P〈0．05），经HBO治疗后SOD活性升高（P〈0．05），MDA含量降低（P〈0．05）。FasLmRNA、caspase-3蛋白在假手术组呈低水平表达，而在IRI组表达显著上调（P〈0．01），IRI＋HBO组表达较IRI组显著下调（P〈0．01）。结论大鼠肾缺血损伤后随着再灌注时间延长FasLmRNA、caspase-3蛋白表达显著上调。早期HBO治疗后可以使FasLmRNA、caspase-3蛋白表达明显下调，抑制细胞凋亡，保护肾脏。Objective To observe the effects of hyperbaric oxygen （HBO） on the expression of FasL mRNA and caspase-3 protein in renal tissue after renal isehemia-reperfusion injury （IRI） in order to elucidate the underlying mechanisms. Methods Rats were randomly divided into three groups： sham group（n=8）, IRI group（n=8） and IRI＋HBO group（n=8）. The IRI group and the IRI＋HBO group recieved 45 minutes hibateral renal isehima and the IRI＋HBO group received additional HBO therapy at the 1st, 24th and 48th hour after ischemia. The kidneys were removed at the end of HBO therapy. Malondialdehyde （MDA） level and superoxide dismutase （SOD） activity were measured to determine the extent of oxidative stress. The expression of FasL mRNA and caspase-3 protein was detected by quantitative real-time PCR and immunohistochemical staining in renal tissue respectively. Results Compared with the sham group, MDA level increased markedly and SOD activity decreased markedly after isehemia. After HBO treatment, MDA level decreased and SOD activity increased significantly （P〈0.05）. In IRI group, the expression of FasL mRNA and caspase-3 protein were higher than those in the sham group （P〈 0.01）, which were reduced significantly by HBO treatment （P 〈0.01）. Conclusion The expression of FasL mRNA and caspase-3 protein increases along with the lasting of reperfusion and HBO exhibites protection against cell apoptosis through improving the antioxidant-oxidant balanceand reducing IRI in acute stage of IRI.

关 键 词：再灌注损伤 细胞凋亡 高压氧 FASL CASPASE-3 

分 类 号：R285.5[医药卫生—中药学]