摘要
目的观察Toll样受体4(TLR4)在胰岛细胞株(INS-1)中的表达情况,探讨内毒素脂多糖(LPS)对INS-1细胞活力、胰岛素分泌功能以及TLR4表达的影响。方法采用RT-PCR技术和Western blotting方法检测INS-1细胞和经1 mg/L LPS刺激的INS-1细胞中TLR4 mRNA和蛋白的表达变化;分别用不同质量浓度的LPS(0.01、0.1、1、5、10 mg/L)刺激INS-1细胞,CCK8法检测细胞活力,GSIS法测定胰岛素分泌功能。结果 RT-PCR和Western blotting检测结果显示:LPS刺激使INS-1细胞TLR4mRNA和蛋白的表达显著增加(P<0.05)。当LPS在0.1～10 mg/L时,INS-1细胞活力受到抑制,且呈剂量时间依赖性(P<0.05);1 mg/L LPS刺激可使高糖环境下的INS-1细胞的胰岛素分泌量显著减少(P<0.05)。结论一定浓度范围内的LPS刺激可抑制INS-1细胞活力,并减少高糖培养条件下细胞胰岛素的分泌量。LPS刺激能上调INS-1细胞TLR4 mRNA和蛋白的表达。内毒素可能通过直接作用损伤胰岛β细胞。
Objective To investigate the expression of Toll-like receptor 4 （TLR4） on INS-1 cells, and explore the effects of lipopolysaccharide （LPS） on viability and insulin secretion of INS-1 cells and expression of TLR4 on INS-1 cells. Methods The expression of TLR4 mRNA and protein in INS-1 cells with or without treatment with 1 mg/L LPS was detected by RT-PCR and Western blotting. After treatment with different mass concentrations of LPS （0.01 mg/L, 0. 1 mg/L, 1 mg/L, 5 mg/L and 10 mg/L）, the viability of INS-1 cells was detected by CCK8 method, and the insulin secretion of INS-1 ceils was determined by GSIS. Results RT-PCR and Western blotting revealed that the expression of TLR4 mRNA and protein in INS-1 cells was significantly increased after treatment with LPS （ P 〈 0.05）. The viability of INS-1 cells was inhibited in a dose and time dependent manner after treatment with 0.1 to 10 mg/L LPS （P 〈0.05）. The insulin secretion of INS-1 ceils was significantly decreased after treatment with 1 mg/L LPS （P 〈 0.05）. Conclusion The viability of INS-1 cells is inhibited, and the insulin secretion function of INS-1 cells is decreased after treatment with LPS of certain concentrations. The expression of TLR4 mRNA and protein in INS-1 cells is increased after treatment with LPS. Pancreatic 13 cells may be damaged by endotoxin in a direct manner.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2012年第12期1577-1580,共4页
Journal of Shanghai Jiao tong University:Medical Science
