CD47过表达MSCs对小鼠脂肪肝缺血再灌注损伤的影响及SIRPα在其中的作用  

Effect of CD47-overexpressed mesenchymal stem cells on fatty liver ischemia-reperfusion injury and role of signal regulatory protein alpha in mice

作　　者：盛明薇[1] 刘伟华[1] 林元邦[2] 喻文立[1] Sheng Mingwei;Liu Weihua;Lin Yuanbang;Yu Wenli(Department of Anesthesiology,Tianjin First Central Hospital,Tianjin 300192,China;Department of General Surgery,Tianjin Medical University General Hospital,Tianjin 300052,China)

机构地区：[1]天津市第一中心医院麻醉科,天津300192 [2]天津医科大学总医院普通外科,天津300052

出　　处：《中华麻醉学杂志》2024年第12期1490-1494,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82370670);天津市卫生健康委员会科技项目(TJWJ2024XK013)。

摘　　要：目的评价CD47过表达间充质干细胞(MSCs)对小鼠脂肪肝缺血再灌注(I/R)损伤的影响及信号调节蛋白α(SIRPα)在其中的作用。方法应用密度梯度离心法分离、培养纯化C57鼠骨髓来源MSCs,将CD47重组腺病毒转染至MSCs;将SPF级4周龄C57小鼠给予高脂饲料诱导中度脂肪肝变性并建立70%肝I/R模型,采用随机数字表法分为4组(n=6):假手术组(S组)、肝I/R组(I/R组)、CD47过表达MSCs组(CD47+I/R组)及CD47过表达MSCs+SIRPα抑制剂组(CD47+RRx+I/R组)。于术前24 h经尾静脉注射MSCs,于术前48 h经尾静脉注射SIRPα抑制剂RRx-0011 mg/kg。于再灌注6 h时麻醉小鼠处死,收集下腔静脉血样和肝组织,采用HE染色法观察小鼠肝病理学结果;测定血清ALT及AST浓度;采用Western blot法检测裂解的半胱天冬酶1(c-caspase-1)、气体衍生物D蛋白(GSDM)及核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)的表达。结果与S组比较,I/R组血清ALT和AST浓度升高,肝组织c-caspase-1、GSDM和NLRP3表达上调(P<0.05),小鼠肝组织病理学损伤加重;与I/R组比较,CD47+I/R组小鼠血清ALT和AST浓度降低,肝组织c-caspase-1、GSDM及NLRP3表达下调(P<0.05),肝组织病理学损伤减轻;与CD47+I/R组比较,CD47+RRx+I/R组小鼠血清ALT和AST浓度升高,肝组织c-caspase-1、GSDM和NLRP3表达上调(P<0.05),肝组织病理学损伤加重。结论CD47过表达MSCs可减轻小鼠脂肪肝缺血再灌注损伤,激活SIRPα抑制细胞焦亡参与了该过程。Objective To evaluate the effect of CD47-overexpressed mesenchymal stem cells(MSCs)on fatty liver ischemia-reperfusion(I/R)injury and the role of signal regulatory protein alpha(SIRPα)in mice.MethodsMSCs from C57 mouse bone marrow were isolated and purified using the density gradient centrifugation,and then CD47 recombinant adenovirus was transfected into MSCs.SPF 4-week-old C57 mice were fed a high-fat diet to induce moderate liver steatosis and then 70%liver I/R model was established.Mice were divided into 4 groups(n=6 each)using a random number table method:sham operation group(S group),liver I/R group(I/R group),CD47-overexpressed MSCs group(CD47+I/R group)and CD47-overexpressed MSCs+SIRPαinhibitor group(CD47+RRx+I/R group).MSCs were injected via the tail vein at 24 h before surgery.SIRPαinhibitor RRx-0011 mg/kg was injected through the tail vein at 48 h before surgery.The mice were anesthetized and sacrificed at 6 h of reperfusion.Blood samples were collected from the inferior vena cava for determination of serum concentrations of alanine transaminase(ALT)and aspartate transaminase(AST).Liver tissues were obtained for examination of the pathological changes and for determiation of the expression of cleaved-caspase 1(c-caspase-1),gasdermin(GSDM)and PYD domains-containing protein 3(NLRP3)(by Western blot).ResultsCompared with S group,the serum concentrations of ALT and AST were significantly increased,the expression of c-caspase-1,GSDM and NLRP3 in liver tissues was up-regulated(P<0.05),and the histo-pathological changes of the liver tissue were aggravated in I/R group.Compared with I/R group,the serum concentrations of ALT and AST were significantly decreased,the expression of c-caspase-1,GSDM and NLRP3 in liver tissues was down-regulated(P<0.05),and the histo-pathological changes of the liver tissue were significantly attenuated in CD47+I/R group.Compared with CD47+I/R group,the serum concentrations of ALT and AST were significantly increased,the expression of c-caspase-1,GSDM and NLRP3 in liver tissues w

关 键 词：间质干细胞 脂肪肝 再灌注损伤 CD47 信号调节蛋白α 

分 类 号：R28[医药卫生—中药学]