脂肪干细胞对大鼠胰岛细胞脂性凋亡的保护作用

Protective Effect of Adipose Derived Stem Cells on Apoptosis of Rat Pancreatic Islet Cells

[目的]探讨脂肪干细胞（ADSCs）对棕榈酸（PA）诱导大鼠胰岛瘤细胞（INS-1）凋亡的保护作用。[方法]本实验分为三组。空白对照组：正常培养液＋INS-1单独培养（其上层只含培养基）；PA组：含0．5mmol／LPA的高脂培养液＋INS-1单独培养；INS-1＋ADSCs组：高脂培养液＋INS-1＋ADSCs共培养。各组在处理24h后检测各组细胞的存活率、胰岛细胞基础胰岛素分泌量（BIS）和葡萄糖刺激的胰岛素分泌量（GSIS）；采用RT-PCR检测胰岛细胞中Ins-1、Glut-2、Pdx-1表达量；采用Elisa检测各组IL-6、TNF—α、HGF、TIMP—1和TGF—β水平。[结果]PA组和PA＋ADSCs组细胞存活率均较对照组下降（P〈0．01），但PA＋ADSCs组细胞存活率明显大于PA组（P〈0．01）。与对照组相比，PA组和PA＋ADSCS组BIS增加（P〈0．01），而GSIS均减少（P〈0．01）；与PA组比较，PA＋ADSCs组细胞BIS差异无统计学意义（P〉0．05），而GSIS显著增加（P〈0．01）。和对照组相比，PA组和PA＋ADSCs组的Pdx-1、Ins-1、Glut-2mRNA水平均明显下降（P〈0．01）；和PA相比，PA＋ADSCs组细胞的Pdx-1和Glut-2mRNA水平明显上升（P〈0．01），而Ins-1mRNA水平比较差异无统计学意义（P〉0．05）。PA＋ADSCs组中IL-6、TNF-α、HGH、TIMP-1、TGF-β的含量均较其他两组明显升高。[结论]PA能诱导INS-1细胞凋亡并抑制Ins-1、Glut-2、Pdx-1的表达，而ADSCs可以通过旁分泌机制并上调Glut-2和Pdx-1的表达对胰岛细胞起到保护作用。

[Objective]To investigate the protective effect of adipose derived stem cells （ADSCs） on the apoptosis of rat pancreatic islet tumor cells （INS-1） induced by palm acid （PA）.[Methods]The experiments were divided into three groups. Blank control group： normal medium ＋INS-1 to be cultured alone （the upper layer contained medium only）; Group PA： high fat medium containing 0.5 mmol/L PA ＋ INS-1 to be cultured alone; INS-1＋ADSCs group： high fat medium ＋ INS-1 ＋ ADSCs for culture. After treatment, The survival rate of the cells, the basal insulin secretion （BIS） and the glucose stimulated insulin secretion （GSIS） were detected after 24 h treatment ; The expression of Ins-1, Glut-2 and Pdx-1 in pancreatic islet cells was detected by RT-PCR; levels of IL-6, TNF-α, HGF, TIMP-1 and TGF-βwere detected by Elisa.[Results]Cell survival rate of PA group and PA＋ ADSCs group was lower than that in control group （ P 〈0.01）, hut the survival rate of the PA＋ADSCs group was significantly higher than that of the PA group （ P 〈0.01）. Compared with the control group, BIS in the PA group and the PA＋ADSCs group increased （ P 〈0.01）, while the GSIS were reduced （ P〈0.01） ; Compared with the PA group, there was no significant difference in BIS cells in the PA＋ADSCs group （ P 〈0.05）, Compared with the control group, the levels of Pdx-1,Ins-1,Glut- 2 mRNA in the PA group and the PA＋ADSCs group were significantly decreased （ P 〈0.01） ; Compared with the PA group, the levels of Pdx-1 and Glut-2 mRNA in the PA＋ADSCs group were significantly increased （ P 〈0.01）;while there was no significant difference in the level of Ins-1mRNA （P 〉0.05）. The contents of IL-6, TNF-α, HGH, TIMP-1 and TGF- in the PA＋ADSCs group were significantly higher than those in the other two groups.[Conclusion] PA can induce the apoptosis of INS-1 cells and inhibit the expression of Ins-1, Glut-2 and Pdx-1. ADSCs can protect the islet cells by the mechanism of side secretion and by the increase of the expression of Glut-2 and Pdx-1.