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.Methods MSCs from C57 mouse bone marrow were isolated and purified using the den-sity 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 oper-ation group(S group),liver I/R group(I/R group),CD47-overexpressed MSCs group(CD47+I/R group)and CD47-overexpressed MSCs+SIRPα inhibitor group(CD47+RRx+I/R group).MSCs were injected via the tail vein at 24 h before surgery.SIRPα inhibitor RRx-001 1 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 col-lected 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).Results Compared 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 sig-nificantly 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 in-creased,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 CD47+RRx+I/R group.Conclusions CD47-overexpressed MSCs can alleviate fatty liver I/R injury in mice,and activation of SIRPα and subse-quent inhibition of pyroptosis are involved in the process.
关键词
间质干细胞/脂肪肝/再灌注损伤/CD47/信号调节蛋白α
Key words
Mesenchymal stem cells/Fatty liver/Reperfusion injury/CD47/Signal regu-latory protein alpha
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