糖尿病大鼠与正常大鼠胰腺干细胞增生能力的比较研究  

Compare Reproductive Activity of Diabetic Rat Pancreatic Stem Cells with that of Normal Rat's Pancreatic Stem Cell

作　　者：张悦[1] 余小舫[1] 张育森[1] 李富荣[1] 周汉新[1] 潘晓华[1] 

机构地区：[1]暨南大学第二临床医学院/深圳市人民医院肝胆胰腺外科

出　　处：《医学研究杂志》2008年第3期52-54,F0003,共4页Journal of Medical Research

基　　金：深圳市科技局科研基金资助项目(200601015)

摘　　要：目的比较糖尿病大鼠与正常大鼠胰腺干细胞的增生能力,探讨大鼠胰岛损伤的微环境对干细胞的影响。方法60mg/kg链脲霉素腹腔注射SD大鼠制备糖尿病大鼠模型为实验组(10只)及正常SD大鼠为对照组(10只),取实验组和对照组胰腺修剪消化成单细胞后接种于培养板中培养;取传代第3代细胞行Nestin免疫组化鉴定;观察细胞生长曲线,取培养至第3代及第7代的细胞以流式细胞术测定细胞周期。结果实验组和对照组大鼠胰腺细胞传代后Nestin免疫组化鉴定证实为胰腺干细胞,糖尿病大鼠胰腺干细胞早期增生能力强[S期细胞所占比率:(20.7±2.7)%vs(14.1±2.5)%,P<0.05],传代时间明显较对照组大鼠短[第3代传代时间:4.6±0.5天vs7.7±0.6天,P<0.05],但第6代以后传代时间趋向一致[第6代传代时间:8.4±0.5天vs8.0±1.0天,P>0.05]。结论大鼠胰腺胰岛损伤后可刺激干细胞增生,干细胞增生能力的变化与微环境有关。Objective Compare the reproductive activity of pancreatic stem cells isolated from diabetic rat with that of normal rat to assess the effect of microenvirement on stem cells＇ proliferation. Methods Diabetic rat model were prepared by peritoneal injecting 60mg/ kg streptozotocin. Pancreata from 10 diabetic rats and 10 normal rats were trimmed and digested into single cells which were then inoculated onto the culture plate respectively. After passaging 3 generation the cells were identified by nestin immunostaining. During culturing the cell growth curve was observed and the cell cycle of the third and seventh generation cultured cells was evaluated by flow cytometry. Resuits After passaging pancreatic stem cells was purified and identified by nestin immunostaining. The reproductive activity of diabetic rat was much better[ ratio ofs stage： （20.7 ±2.7）% vs（14.1±2.5）%,P 〈0.05],and the generation time of diabetic rats were much shorter than that of normal at the beginning[ GT of 3 rd generation ： （ 4.6 ± 0.5 ） d vs （ 7.7 ± 0.6 ） d, P 〈 0.05 ] , and after passaging 6 generations the generation time trend to the same[ GT of 6th generation ： 8.4 ± 0.5d vs 8.0 ±1.0d,P 〉 0.05 ]. Conclusions The stem cells are stimulated to proliferate after islet injure. The reproductive activity of cells can be provoked by the microenvironment.

关 键 词：糖尿病 胰腺干细胞 增生能力 微环境 

分 类 号：R734.2[医药卫生—肿瘤] R587.1[医药卫生—临床医学]