干细胞来源外泌体对大鼠肝脏缺血再灌注损伤的保护作用  

Protective effect of exosomes from stem cells on liver ischemia-reperfusion injury in rats

作　　者：何其宽 戴宁高 叶瑞凡 洪重 余华军[1] 郭鹏毅[1] 张杰[1] 张启瑜[1] HE Qi-kuan;DAI Ning-gao;YE Rui-fan;Hong Zhong;Yu Hua-jun;Guo Peng-yi;Zhang Jie;Zhang Qi-yu(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou,Zhejiang 325000,China;Department of Hepatobiliary Surgery,the Second Affiliated Hospital of Wenzhou Medical University,Wenzhou,Zhejiang 325000,China)

机构地区：[1]温州医科大学附属第一医院肝胆外科,浙江温州325000 [2]温州医科大学附属第二医院肝胆外科,浙江温州325000

出　　处：《肝胆胰外科杂志》2018年第2期134-141,共8页Journal of Hepatopancreatobiliary Surgery

基　　金：浙江省自然科学基金资助项目(LY13H030009);国家自然科学基金资助项目(81470874)

摘　　要：目的研究大鼠脂肪间充质干细胞来源的外泌体(exosomes,Exo)对肝脏缺血再灌注损伤(hepatic ischemia-reperfusion injury,HIRI)的保护作用。方法胶原酶法提取SD大鼠原代脂肪间充质干细胞,超速离心法提取干细胞来源的外泌体。SD大鼠随机分为假手术组(Sham)、缺血再灌注损伤组(IR)、缺血再灌注损伤外泌体治疗组(IR+Exo)。Sham组腹部正中切口,离断肝脏周围韧带,不做其他处理;IR组用动脉夹阻断70%肝脏血流60 min;IR+Exo组70%肝脏血流阻断60 min后,尾静脉注射外泌体观察其对肝脏缺血再灌注损伤的保护作用。检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)含量。检测肝组织中超氧化物歧化酶(SOD)与丙二醛(MDA)含量。Western blotting法检测氧化应激指标醌氧化还原酶(NQO-1)、血红素氧合酶1(HO-1),凋亡相关指标Bcl-2、Bax,HE染色观察肝组织病理变化。结果相比IR组,IR+Exo组血清中ALT、AST、TNF-α、IL-1β水平显著下降(P<0.05),SOD活性显著升高(P<0.05),MDA含量明显下降(P<0.05)。NQO1、HO-1、Bcl-2表达量显著升高,Bax显著下降(P<0.05),肝组织病理损伤明显改善。结论外泌体能够减轻肝脏缺血再灌注损伤,其可能通过缓解氧化应激、抑制炎症反应、抗凋亡而发挥作用。Objective To investigate the protective effect of exosomes(Exo)from adipose derived mesenchymal stem cells on hepatic ischemia-reperfusion injury(HIRI)in rats.Methods Adipose derived mesenchymal stem cells were extracted from Sprague-Dawley(SD)rats in collagen enzymatic way.Exosomes were isolated from conditioned culture medium by ultracentrifugation.SD rats were randomly divided into Sham group,IR group and IR+Exo group.Exosomes were injected via the tail vein in male SD rats model of 70%hepatic IR injury,and the therapeutic effect was evaluated.The serum levels of AST,ALT,TNF-αand IL-1βwere measured by enzyme linked immunosorbent assay(ELISA).SOD activity and MDA level were measured in liver tissues.The expression of oxidative stress indexes NQO-1,HO-1,apoptosis indexes Bcl-2,Bax were assessed by Western blotting.Hematoxylin-eosin staining was used to observe the histopathological changes.Results Compared with the IR group,the activities of ALT,AST,TNF-αand IL-1βin IR+Exo group were significantly decreased(P<0.05),SOD activity increased and MDA level decreased considerably(P<0.05).NQO-1,HO-1,Bcl-2 were noticeably increased,Bax decreased remarkably(P<0.05).The histopathological damages were significantly improved in IR+Exo group.Conclusion Exosomes can attenuate hepatic ischemia-reperfusion injury and may play a role in suppressing inflammatory reaction,alleviating oxidative stress and anti-apoptosis.

关 键 词：脂肪间充质干细胞 外泌体 肝脏缺血再灌注损伤 炎症反应 氧化应激 凋亡 大鼠 

分 类 号：R575[医药卫生—消化系统]