健脾补肾清肠化湿方对静脉移植MSCs向溃疡性结肠炎大鼠结肠迁移分化的影响  

Effects of Jianpi Bushen Qingchang Huashi Decoction on Migration and Differentiation of MSCs in Ulcerative Colitis Rat

作　　者：朱磊[1,2] 沈洪 刘丽 顾培青 王玖[2] 蒋寅[2] 朱长乐 司海鹏 朱萱萱 

机构地区：[1]南京中医药大学附属医院(江苏省中医院),南京210029 [2]南京中医药大学,南京210029

出　　处：《中国实验方剂学杂志》2015年第21期88-92,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81403344;81373606);国家中医药行业科研专项项目(201407001);国家中医药管理局项目(JDZX2012079);江苏省临床医学科技专项项目(BL2014100);江苏高校优势学科建设工程项目(PAPD);国家中医临床研究基地(脾胃)

摘　　要：目的:观察健脾补肾清肠化湿方对静脉移植的大鼠骨髓间充质干细胞(MSCs)向溃疡性结肠炎模型大鼠结肠迁移归巢的影响。方法:体外分离培养SD大鼠MSCs,传至第3代,制备细胞悬液,加入0.78 mg·L-1健脾补肾清肠化湿方共培养,并以4',6-二脒基-2-苯基吲哚(4',6-diamidino-2-phenylindole,DAPI)荧光标记备用。大鼠分为空白组,模型组,MSCs组,干预MSCs组和联合组。2,4,6三硝基苯磺酸(TNBS)法建立溃疡性结肠炎大鼠模型,空白组、模型组大鼠尾静脉分别iv 1 m L生理盐水,MSCs组大鼠,iv 1 m L MSCs(1×106个),干预MSCs和联合组大鼠分别尾静脉iv体外中药干预的MSCs(1×106个),联合组再予健脾补肾清肠化湿方13.6 g·kg-1,ig 10 d。第5,10天每组各处死5只大鼠,结肠标本进行大体形态和组织学评估,共聚焦显微镜观察各组DAPI标记MSCs在结肠内迁移情况,免疫组化检测肠干细胞标记物神经细胞RNA结合蛋白(Musashi-1)的表达。结果:与正常组比较,模型组大鼠结肠可见广泛充血水肿,伴糜烂,肉眼可见溃疡形成,提示造模成功。除模型组和正常组外,各移植组结肠组织均能检测到DAPI标记的MSCs,且数量随时间的延长而逐渐增加;各治疗组均能改善大鼠结肠组织大体形态和组织病理,且第10天联合组优于MSCs组(P<0.05);各移植组能提高Musashi-1的表达,第10天联合组和干预MSCs组优于MSCs组(P<0.05)。结论:健脾补肾清肠化湿方能促进MSCs向结肠溃疡部位的迁移和分化,并能修复受损的结肠黏膜。Objective： To observe whether Jianpi Bushen Qingchang Huashi decoction（ JBQHD） can promote the migration of mesenchymal stem cells（ MSCs） intravenously transplanted in rat＇s bone to the colon of ulcerative colitis（ UC） rat model. Method： MSCs were isolated and cultured in vitro,then spread to the third generation to prepare cell suspension,and 0. 78 mg·L- 1JBQHD was added into it for co-culture. Fluorescent prelabeling by 4＇,6-diamidino-2-phenylindole（ DAPI） was carried out in vitro. The rats were divided into blank group,model group,MSCs group,intervened MSCs group and combined group. UC rats models were established by TNBS method. The rats of blank group and model group received intravenous injection with 1 m L normal saline respectively through tail veins. The rats of MSCs group received intravenous injection with 1 m L MSCs through tail veins（ 1 × 106CFU）. The rats of intervened MSCs group and combined group received intravenous injection with MSCs that had been intervened in vitro with the decoction（ 1 × 106CFU）,and the combined group also received13. 6 g·kg- 1JBQHD by ig for 10 days. Five rats were killed respectively in each group on Day 5 and Day 10. Gross morphological and histological evaluation was taken for the colon specimens,and confocal microscope was used to observe the migration of MSCs labeled by DAPI in the colon tissues; immunohistochemical staining was conducted to detect the expression of Musashi-1（ a marker of intestinal stem cells）. Result： Compared with the normal group,the rats in model group had extensive hyperemia and edema in colon tissues,accompanied by erosion and visible ulceration,indicating successful modeling. Except model group and normal group,MSCs labeled by DAPI can be detected in colon tissues in all other groups,and the quantity increased gradually with time. The all treatment groups could improve gross morphology and histopathology in colon tissues of rats,and the result in combined group was superior to that in MSCs group（ P �

关 键 词：溃疡性结肠炎 健脾补肾 清肠化湿 骨髓间充质干细胞 移植 迁移 

分 类 号：R285.5[医药卫生—中药学]