神经元素3在不同纯度供体胰岛细胞中的表达

Expression of Ngn3 in the donor islets with different purity

目的:研究神经元素3(Ngn3)在不同纯度胰岛细胞中的表达情况,探讨不同纯度胰岛细胞中所携带的干细胞。方法:30只成年雄性SD大鼠随机分为3组,采用胰管内注入胶原酶消化法分离成年大鼠胰腺,Ⅰ组(n=10):采用2次消化、不纯化的方法获得胰岛;Ⅱ组(n=10):胰岛沉淀物加入25%Ficoll-400纯化,Ⅲ组(n=10):胰岛沉淀物依次加入25%、11%的Ficoll-400纯化。应用免疫组化、逆转录-多聚酶链反应(RT-PCR)方法进行定位、半定量分析,检测Ngn3在不同纯度胰岛细胞中的表达。结果:Ⅰ组获得胰岛纯度约为43.6%±6.29%,Ⅱ组获得胰岛纯度约为65.3%±4.40%,Ⅲ组获得胰岛纯度约为77.6%±6.36%,各组比较均具有显著性差异(P<0.05)。Ngn3在不同纯度胰岛细胞中均有表达,Ⅰ组中Ngn3的蛋白和和核酸水平表达明显高于Ⅱ组、Ⅲ组,Ⅲ组中表达最弱,各组比较均具有显著性差异(P<0.05)。结论:Ngn3在不同纯度胰岛细胞中均有表达,在低纯度胰岛细胞中表达最明显,提示低纯度胰岛细胞中可能携带较多的干细胞。

Objective：To study the expression of Ngn3 in rats donor islets with different purity,and explore the stem cells in them.Methods：Thirty male adult SD rats were randomly divided into 3 groups.Islets were isolated by digestion method of injecting collagenase into pancreatic duct.Group Ⅰ was harvested by two digestions without purification;For group Ⅱ （n=10）,islets sediments were purified by adding 25% ficoll-400 and for,Group Ⅲ（n=10）,by adding （25% and 11%）ficoll-400 in order.Finally we applied immunohistochemisiry and RT-PCR to the location and semi-quantitative analysis to test the expression of Ngn3 in the donor islets with different purity.Results：Islets purity of group Ⅰ was 43.6%±6.29%,Group Ⅱ 65.3%±4.40%,and group Ⅲ 77.6%±6.36%.The difference of Islet purity was significant in three groups （P〈0.05）.Compared with group Ⅱ and group Ⅲ,the expressions of protein and mRNA of Ngn3 were both highest in group Ⅰ,lowest in group Ⅲ.（P〈0.05）.Conclusion：Ngn3 was expressed in all islets,with the low purity expressing the most prominent,which suggests that islets with low purity may bear more stem cells.