摘要
[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.
出处
《医学临床研究》
CAS
2016年第10期1911-1914,共4页
Journal of Clinical Research