钨酸钠促进新生猪胰岛细胞体外增殖、分化及分泌功能

Sodium tungstate promotes the proliferation,differentiation and secretion of neonatal porcine islet cells in vitro

目的观察钨酸钠在体外对新生猪胰岛细胞增殖及功能活性的影响。方法分离和纯化新生猪胰岛细胞后,与钨酸钠共育,隔天进行细胞计数、MTT实验,取最佳浓度300μmol/L钨酸钠与细胞共同培养3d,收集细胞进行葡萄糖刺激实验,并分别用Western blot、RT-PCR方法检测细胞内胰岛素蛋白(INSULIN)、细胞增殖核抗原(PCNA)和胰十二指肠同源框蛋-1(PDX-1)mRNA、葡萄糖转运子-2(GLUT-2)mRNA的表达。结果 300μmol/L钨酸钠处理后胰岛细胞增殖活性及葡萄糖刺激实验中胰岛素分泌明显高于对照组(P<0.05),钨酸钠干预组PCNA蛋白和IN-SULIN蛋白表达分别为2.24±0.19和1.62±0.17,显著高于对照组的1.17±0.13和0.37±0.08(P<0.05,P<0.01);PDX-1和GLUT-2 mRNA表达分别为0.34±0.05和1.06±0.10,显著高于对照组的0.11±0.03和0.13±0.02(P<0.01)。结论低浓度(300μmol/L)钨酸钠具有促进新生猪胰岛细胞增殖分化、增强胰岛细胞功能、促进胰岛素分泌的作用。

Objective To investigate the possible mechanism of sodium tungstate in diabetic therapeutical effects and the protective function in neonatal porcine islet cells in vitro, the effects of sodium tungstate on proliferation and differentiation of neonatal porcine islet cells was observed in vitro in our experiment. Methods Neonatal porcine islets （NPIS） were cultured with various concentrations of sodium tungstate and carry out cell counting and MTT assay every other day. NPIS was exposed to the most proper concentration 300 txmol/L sodium tungstate tbr three days, and the glucose-stimulated insulin secrtion （GSIS） were detected, the expression of proliferating cell nuclear antigen （PCNA） , insulin content in islet cells and PDX-1 mRNA, GLUT-2 mRNA were assayed by Western blot and RT-PCR respectively. Results The group of islet cells were cultured with the dose of 300 μmol/L had the higher light absorption in MTF assay （P 〈 0.05 ） and GSIS was significantly increased in the group. The protein expressions of PCNA and INSULIN content in the experimental group were 2. 24 ± 0. 19 and 1.62 ± 0. 17 signifi- cantly higher than that in control group which were 1.17 ±0. 13 and 0. 37 ±0. 08 （P 〈0. 05 ,P 〈0. 01 ）. The mRNAexpressions of PDX-1 and GLUT-2 were 0. 34 ± 0. 05 and 1.06 ± 0. 10 in the experimental group significantly higher than that in control group which were 0. 11 ± 0. 03 and O. 13 ± 0. 02 （P 〈 0. 01 ）. Conclusions 300μmol/L sodium tungstate has the advanced effect on promoting the proliferation and differentiation of neonatal porcine islet cell. It also has the effect in enhancing the function of insulin secretion.