摘要
目的:观察健脾补肾、清肠化湿方联合骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)对溃疡性结肠炎模型大鼠结肠黏膜的修复作用.方法:SD大鼠随机分为空白组、模型组、BMSCs组、干预BMSCs组和联合组.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注射1 mL生理盐水,BMSCs组大鼠尾静脉注射BMSCs,干预BMSCs和联合组大鼠分别注射体外中药共培养的BMSCs,联合组再予中药13.6 g/kg灌胃10 d.分别于第5天和第10天每组各处死5只大鼠,留取标本.电子显微镜观察大鼠结肠组织杯状细胞数量,实时定量PCR检测(real-time quantitative PCR,qRT-PCR)和Western blot法分别检测大鼠结肠组织Muc2mRNA和蛋白的表达,Western blot法检测Muc2相关因子Math1和KLF-4蛋白的表达.结果:与模型组相比,各治疗组肠黏膜杯状细胞数量增多,各治疗组Muc2 mRNA水平升高,联合组优于干预BMSCs组和BMSCs组(P<0.05).随着时间延长,治疗效果越明显.治疗第10天,与正常组相比,模型组Muc2、Math1和KLF-4蛋白表达量明显下降,经治疗后,各治疗组蛋白表达量均上高,联合组升高最明显.结论:静脉移植BMSCs的同时给予健脾补肾、清肠化湿方能提高黏蛋白Muc2表达水平,增多杯状细胞数量,修复肠屏障,联合灌胃持续给药效果更佳,其机制可能和提高Math1和KLF-4因子表达有关.
AIM：To observe the effect of traditional Chinese compound medicine Jianpi Bushen Qingchang Huashi Decoction combined with bone mesenchymal stem cells（BMSCs） in the repair of the intestinal mucosa of rats with ulcerative colitis.METHODS：Rats were divided into five groups：a normal group,a model group,a BMSCs group,an intervened BMSCs group and a combination group.The rats of the normal and model groups received an intravenous injection of normal saline separately through the tail vein.The BMSCs group received an intravenous injection of BMSCs（1×10~6/mL）through the tail vein.The intervened BMSCs group and combination group received BMSCs（1×10~6/mL） intervened by decoction in vitro,and the combination group additionally received the oral decoction for 10 days.Five rats were killed on the 5^（th） and 10^（th） day after the transplantation,respectively.The mRNA and protein expression of Muc2 was detected by real-time PCR and Western blot,respectively.Expression of Math1 and KLF-4 was assayed by Western blot.RESULTS：The number of goblet cells was increased in each BMSCs transplantation group.The mRNA expression of Muc2 significantly increased in each treatment group relative to the model group,and the increase was more significant in the combination group than in the intervened BMSCs and BMSCs groups.As time increased,the therapeutic effect was more obvious.On the 10 th day of treatment,compared with the normal group,Muc2,Mathl and KLF-4protein expression was significantly decreased in the model group,while the expression of these proteins was higher in the treatment groups,with the combination group increasing most obviously.CONCLUSION：Transplantation of BMSCs combined with traditional compound Chinese medicine could improve the mRNA and protein expression of Muc2 in UC rats,reduce inflammation and repair intestinal barrier,which may be related to the Math1 and KLF-4 factors.
出处
《世界华人消化杂志》
CAS
2016年第7期1017-1023,共7页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目,No.81373606
国家中医药行业科研专项基金资助项目,No.201407001
江苏省临床医学科技专项基金资助项目,No.BL2014100~~
