间充质干细胞对回肠贮袋炎大鼠的治疗作用  

The therapeutic efficacy of mesenchymal stem cells on ileal pouchitis of rats

作　　者：余子璇 李凯钰 何安琪[1] 贾锦滟 胡芳[2] 刘刚[1] Yu Zixuan;Li Kaiyu;He Anqi;Jia Jinyan;Hu Fang;Liu Gang(Department of General Surgery,General Hospital of Tianjin Medical University,Tianjin 300052,China;Department of Nursing,General Hospital of Tianjin Medical University,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院普通外科,天津300052 [2]天津医科大学总医院护理部,天津300052

出　　处：《中华实验外科杂志》2024年第3期458-463,共6页Chinese Journal of Experimental Surgery

摘　　要：目的探讨人脐带间充质干细胞(hUC-MSCs)对全结直肠切除、回肠储袋肛管吻合术(IPAA)术后大鼠回肠贮袋炎模型的治疗作用及其机制。方法20只雄性Sprague-Dawley大鼠建立回肠贮袋炎模型,建模成功后随机数字表法选取2只用于干细胞示踪,剩余18只大鼠随机数字表法分为间充质干细胞组(MSC组)、磷酸盐缓冲液组(PBS组)和葡聚糖硫酸钠组(DSS组)各6只,分别予2×10^(6) hUC-MSCs腹腔注射1次、1 ml PBS腹腔注射1次、无特殊干预。记录各组大鼠体重及粪便评分变化、血便情况、疾病活动指数(DAI)评分,于给药4 d后处死。取贮袋标本进行组织病理学评分,髓过氧化物酶(MPO)表达水平检测,白细胞介素(IL)-1β、IL-6、IL-10、肿瘤坏死因子-α(TNF-α)mRNA相对表达量和蛋白表达量检测。正态分布资料的两组间比较采用t检验;非正态分布资料和等级资料的两组间比较采用Mann-Whitney U秩和检验;多组独立样本间的比较采用Kruskal-Wallis H秩和检验。结果MSC经腹腔注射后贮袋部位聚集明显,荧光显影于注射4 d后消失。MSC组大鼠体重开始回升,血便症状消失;而PBS组和DSS组大鼠体重趋缓但仍持续下降,血便症状未见好转。干细胞治疗4 d后MSC组大鼠DAI低于PBS组和DSS组(0.89±0.34比2.95±0.25,Z=-2.966,P<0.01;0.89±0.34比2.78±0.17,Z=-2.956,P<0.01);MSC组组织病理学评分低于PBS组和DSS组(1.50±0.84比8.17±1.33,Z=-2.989,P<0.01;1.50±0.84比8.33±1.51,Z=-2.956,P<0.01);MPO蛋白表达量低于PBS组和DSS组(0.02±0.00比0.07±0.00,Z=-2.892,P<0.01;0.02±0.00比0.08±0.00,Z=-2.903,P<0.01);MSC组IL-1β、IL-6、TNF-α的mRNA相对表达量低于PBS组和DSS组(0.02±0.00比0.06±0.02,0.02±0.00比0.06±0.02,Z=-2.882,P<0.01;0.05±0.01比0.12±0.01,0.05±0.01比0.11±0.01,Z=-2.882,P<0.01;0.04±0.00比0.06±0.01,0.04±0.00比0.06±0.01,Z=-2.882,P<0.01);同时IL-1β、IL-6、TNF-α蛋白表达量(pg/mg)也低于PBS组和DSS组(0.73±0.04比1.40±0.32,0.73±0.04比1.41±0Objective To investigate the therapeutic effect and mechanism of human umbilical cord mesenchymal stem cells(hUC-MSCs)on rat model of ileal pouchitis after total colorectal resection and ileal pouch anal anastomosis(IPAA).MethodsThe ileal pouchitis models were established in 20 male Sprague-Dawley rats.After successful modeling,2 rats were randomly selected for stem cell tracing.The remaining 18 rats were randomly divided into MSC group,phosphate buffer salt solution group(PBS group)and dextrosan sulfate sodium group(DSS group)with 6 rats each.A total of 2×10^(6) hUC-MSCs were intraperitoneally injected once,and 1 ml PBS was intraperitoneally injected once,without special intervention.The changes of body weight,fecal score,blood stool status and disease activity index(DAI)score of each group were recorded,and the rats were killed after 4 days of administration.The bag samples were collected for histopathological score,myeloperoxidase(MPO)expression level,interleukin(IL)-1β,IL-6,IL-10,tumor necrosis factor-α(TNF-α)mRNA expression and protein expression.The comparison between the two groups of normal distribution data was performed by t test.Mann-Whitney U rank sum test was used to compare the non-normal distribution data and rank data between the two groups.Kruskal-Wallis H-rank sum test was used to compare multiple independent samples.ResultsThe concentration of MSCs in the storage bag was obvious after intraperitoneal injection,and the fluorescence disappeared at 4th day after injection.The weight in MSC group began to recover,and the symptoms of blood and stool disappeared.However,the body weight of rats in PBS group and DSS group slowed down but continued to decline,and the symptoms of blood and stool were not improved.After 4 days of stem cell therapy,DAI in MSC group was significantly lower than that in PBS group and DSS group(0.89±0.34 vs.2.95±0.25,Z=-2.966,P<0.01;0.89±0.34 vs.2.78±0.17,Z=-2.956,P<0.01).Histopathological scores in MSC group were significantly lower than those in PBS group and DSS gr

关 键 词：贮袋炎 人脐带间充质干细胞 回肠贮袋肛管吻合术 大鼠 

分 类 号：R656[医药卫生—外科学]