骨髓间充质干细胞对重症急性胰腺炎胰腺组织修复的促进作用研究  

Positive effect of bone marrow-derived mesenchymal stem cells on the repair of pancreatic tissue in severe acute pancreatitis

作　　者：钱道海 宋国栋 张洲 马志龙 王冠男[1] 于文建 陈鹏[1] 王小明[1] Qian Daohai;Song Guodong;Zhang Zhou;Ma Zhilong;Wang Guannan;Yu Wenjian;Chen Peng;Wang Xiaoming(Hepato-biliary Surgery,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China;Hepato-biliary Surgery,the Tenth People's Hospital,Affiliated to Tongji University,Shanghai 270002,China)

机构地区：[1]安徽省芜湖市皖南医学院附属弋矶山医院肝胆外科,芜湖241001 [2]同济大学附属第十人民医院肝胆外科,上海270002

出　　处：《中华细胞与干细胞杂志（电子版）》2019年第6期351-357,共7页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：安徽省教育厅高校自然基金重点项目（KJ2017A271）;安徽省中央引导地方科技发展专项基金（YDZX20183400004899）;弋矶山医院引进人才科研基金（YR201601）;弋矶山医院科研能力“高峰”培育计划项目（GF2019G03）

摘　　要：目的观察骨髓间充质干细胞(BMSCs)抑制坏死性凋亡促进修复重症急性胰腺炎(SAP)的可能机理。方法(1)分离、培养和鉴定大鼠BMSCs;(2)构建牛磺胆酸钠(NaT)诱导的SAP大鼠模型,并分成正常组(NC)、假手术组(Sham)、SAP模型组(SAP)、PBS治疗组(PBS)、BMSCs治疗组和Necrostain-1(Nec-1)治疗组,并检测胰腺病理评分和血淀粉酶水平;(3)运用Western Blot和qRT-PCR方法检测各组受损胰腺组织内RIPK1、RIPK3、Caspase-8、MLKL蛋白及mRNA表达,各组均数间比较采用单因素方差分析,两两比较采用LSD-t检验。结果BMSCs可以被诱导分化成骨、软骨、脂肪,并高表达CD44(99.82)、CD73(99.87)、CD90(99.99)、CD105(99.78),低表达CD11b(0.65)、CD19(0.85)、CD34(0.70)和CD45(1.20)。SAP组胰腺病理评分(12.90±1.79)及血淀粉酶水平(1052.41±183.12)mU/ml均高于NC组[评分:0.40±0.52,淀粉酶水平:(236.62±33.21)mU/ml]和Sham组[评分:0.50±0.53,淀粉酶水平:(242.31±27.94)mU/ml](F=200.275,F=143.245,P均<0.001),且SAP组受损胰腺组织内RIPK1、RIPK3、MLKL表达升高、Caspase-8表达降低(F=179.905,P<0.001);BMSCs组和Nec-1治疗组胰腺病理评分及血淀粉酶水平均低于PBS治疗组[评分:7.20±1.23、7.00±1.05比12.60±1.65,F=200.275,P<0.001;淀粉酶水平:(452.21±101.68)mU/ml、(570.18±148.47)mU/ml比(972.77±204.29)mU/ml,F=143.245,P<0.001],同时受损胰腺组织内RIPK1、RIPK3、MLKL表达下调、Caspase-8表达升高(F=179.905,P<0.001)。结论BMSCs可能通过抑制坏死性凋亡通路活化来修复SAP。Objective To investigate the possible mechanism of bone marrow mesenchymal stem cells(BMSCs)promoting the repair of severe acute pancreatitis(SAP)by inhibiting necropotosis.Methods(1)the isolation,culture and identification of rat BMSCs;(2)the rat SAP models induced by sodium sulfonycholate(NaT)were constructed and divided into the normal group(NC),Sham group(Sham),SAP model group,PBS treatment group,BMSCs transplantation group and the Necrostain-1(Nec-1)treatment group.(3)The Western-blot and qRT-PCR were used to detect the protein and mRNA expressions of RIPK1,RIPK3,caspase-8 and MLKL in the damaged pancreatic tissues of each group.One-way anova was used for comparison between the mean values of each group,and LSD-t test was used for pair comparison.Results BMSCs could be induced to differentiate into bone,cartilage and fat,with high expressions of CD44(99.82﹪),CD73(99.87﹪),CD90(99.99﹪)and CD105(99.78﹪),low expressions of CD11b(0.65﹪),CD19(0.85﹪),CD34(0.70﹪)and CD45(1.20﹪).Pancreatic pathology scores(12.90±1.79)and blood amylase level(1052.41±183.12)mU/ml in SAP model group were significantly higher than those in NC group[scores:0.40±0.52,amylase level:(236.62±33.21)mU/ml]and Sham group[scores:0.50±0.53,amylase level:(242.31±27.94)mU/ml](F=200.275,F=143.245,P<0.001).Moreover,the expressions of RIPK1,RIPK3 and MLKL were significantly increased and the expression of caspase-8 was significantly decreased in the SAP model group(F=179.905,P<0.001).Pancreatic pathology scores and blood amylase level in BMSCs transplantation group and Nec-1 treatment group were significantly lower than those in PBS treatment group[score:7.20±1.23,7.00±1.05 vs 12.60±1.65,F=200.275,P<0.001;Amylase levels:(452.21±101.68)mU/ml,(570.18±148.47)mU/ml vs(972.77±204.29)mU/ml,F=143.245,P<0.001],while the expressions of RIPK1,RIPK3,MLKL were significantly down-regulated and the expression of caspase-8 was significantly increased in the damaged pancreas(F=179.905,P<0.001).Conclusion BMSCs may repair SAP by inhibiting t

关 键 词：坏死性凋亡 CD11B 骨髓间充质干细胞 CD45 CD90 胰腺组织 单因素方差分析 重症急性胰腺炎 

分 类 号：R73[医药卫生—肿瘤]