脐带间充质干细胞来源的外泌体对大鼠创伤性胰腺炎损伤的修复  

The repair effect of exosomes derived from human umbilical cord mesenchymal stem cells on traumatic pancreatitis in rats

作　　者：韩丽 陈思平 赵治戎 周世波 周黎晨 吉华 唐政 谭震 王涛 戴睿武 HAN Li;CHEN Siping;ZHAO Zhirong;ZHOU Shibo;ZHOU Licheng;JI Hua;TANG Zheng;TAN Zhen;WANG Tao;DAI Ruiwu(Faculty of Clinical Medicine,College of Medicine,Southwest Jiaotong University,Chengdu,610031,Sichuan Province;Department of General Surgery,General Hospital of Western Theater Command,Chengdu,610083,Sichuan Province,China)

机构地区：[1]西南交通大学医学院临床医学系,成都610031 [2]西部战区总医院普通外科,成都610083

出　　处：《陆军军医大学学报》2022年第16期1642-1650,共9页Journal of Army Medical University

基　　金：西部战区总医院院管课题(2021-XZYG-B16);四川省科技厅重点研发项目(2022YSFS0195)。

摘　　要：目的探讨脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)来源的外泌体(hUC-MSCs exosome,hUCMSC-Ex)对创伤性胰腺炎的治疗和保护作用。方法60只成年雄性健康SD大鼠(体质量200~250 g),通过随机区组法分为4组(n=15):对照组(Control组),开腹后轻轻翻动胰腺,随后关腹,经尾静脉注射1 mL生理盐水;创伤性胰腺炎组(TP组),使用造模仪建立创伤性胰腺炎大鼠模型,经尾静脉注射1 mL生理盐水;TP+hUC-MSCs组(造模后经尾静脉注射干细胞)和TP+hUCMSC-Ex组(造模后经尾静脉注射外泌体)。采用超速离心法获取外泌体,使用透射电镜对外泌体进行形态学鉴定;HE染色观察胰腺组织损伤的程度;ELISA法检测淀粉酶、脂肪酶、促炎因子、抗炎因子的表达水平;原位末端标记法(TUNEL)检测细胞凋亡情况;Western blot检测细胞凋亡蛋白Bax、Caspase-3表达水平。结果与TP组比较,TP+hUC-MSCs组和TP+hUCMSC-Ex组的胰腺组织病理学评分明显降低(P<0.05),血清淀粉酶、脂肪酶浓度、IL-6、TNF-α表达水平明显降低(P<0.05),而IL-10、TGF-β表达水平明显升高(P<0.05)。TP组的凋亡指数明显高于对照组(P<0.05),而TP+hUC-MSCs组和TP+hUCMSC-Ex组的凋亡指数显著低于TP组(P<0.05),且TP+hUC-MSCs组和TP+hUCMSC-Ex组的凋亡蛋白Bax、Caspase-3表达水平明显降低(P<0.05)。结论hUCMSC-Ex可以通过控制全身性的炎症反应和抑制大鼠腺泡细胞凋亡,参与胰腺组织再生修复。Objective To explore the curative and protective effects of exosomes derived from human umbilical cord mesenchymal stem cells(hUCMSC-Ex)on traumatic pancreatitis in rats.Methods Sixty grown male SD rats(200~250 g)were selected and randomly divided into four groups(n=15 of each group):control,traumatic pancreatitis(TP),TP+hUC-MSCs,and TP+hUCMSC-Ex groups.In the control group,the pancreas was gently turned over after laparotomy.Subsequently,the abdomen was closed and 1 mL normal saline was injected via the tail vein.In the TP group,a rat model of TP was established using a model impactor and 1 mL of normal saline was injected via the tail vein.hUC-MSCs or hUCMSC-Ex were injected via tail vein after modeling in TP+hUC-MSCs and TP+hUCMSC-Ex groups.The ultracentrifugation was used to obtain exosomes,and transmission electron microscopy was used to identify exosomes;HE staining was used for detecting the severity of damage of pancreatic tissue;The expression levels of amylase,lipase,pro-inflammatory factors and anti-inflammatory factors were detected by ELISA;Cell apoptosis was detected by TUNEL;The expression levels of apoptosis proteins Bax and Caspase-3 were detected by Western blotting.Results Compared with TP group,TP+hUC-MSCs group and TP+hUCMSC-Ex group had obvious lower score on pancreas histopathological scoring(P<0.05);The concentrations of serum amylase and lipase were significantly decreased(P<0.05);The expression levels of serum pro-inflammatory factors IL-6 and TNF-αwere significantly decreased,while the expression levels of anti-inflammatory factors IL-10 and TGF-βwere significantly increased(P<0.05);The apoptotic index in the TP group was significantly higher than that of in the control group(P<0.05),and the apoptotic index in the TP+hUC-MSCs group and TP+hUCMSC-Ex group was significantly lower than that in the TP group(P<0.05).Compared with TP group,the expression levels of pancreatic apoptotic proteins Bax and Caspase-3 were significantly decreased in the TP+hUC-MSCs group and TP+hUCMSC-Ex group(P<0

关 键 词：外泌体 脐带间充质干细胞 创伤性胰腺炎 凋亡 

分 类 号：R-332[医药卫生] R329.24R657.51