目的:评估高糖对肾组织干细胞(KSC)基本特征的影响。方法分离肾乳头处KSC,比较其在高糖(30 mmol/L)和正糖(5.6 mmo l/L)培养基的微环境下细胞形态、增殖能力、相关表型标记物的改变。结果细胞增殖检测结果提示从第4天开始高糖组细胞生长较正糖组缓慢;免疫荧光结果提示KCS经高糖处理后,纤维连接蛋白、Ⅰ型胶原、Ⅳ型胶原的合成增加;qRT-PCR检测结果显示KSC经高糖干预后,纤维连接蛋白、Ⅰ型胶原、Ⅳ型胶原mRNA 表达增高,间质标记物α-SMA、Vimentin基因表达进一步升高,促纤维化因子TGF-β、CTGF、PAI-1 mRNA表达均升高。结论 KSC在高糖的微环境中细胞增殖能力下降,促纤维化基因表达增高。
Objective To evaluate the effects of high glucose on the basic features of kidney stem cells. Methods Isolated the KSC from the renal papilla in 4 weeks-Sprague-Dawley rats, the cells were cultured in the media with high glucose (30 mmol/L) and with normal glucose (5.6 mmol/L) respectively. The changes of the cell morphology, proliferation, phenotypic markers were investigated. Results KSC grew slowly in the high glucose media. Immunofluorescence showed that, the synthesis of fibronectin, collagenⅠ and collagen Ⅳ increased after KSC cultured in high glucose media. And qRT-PCR test indicated that the expression of fibronectin, collagenⅠ and collagen Ⅳ mRNA also increased. The gene expression of mesenchymal marker α-SMA, Vimentin and fibrosis factors marker TGF-β, CTGF, PAI-1 were higher than that cultured in normal media. Conclusion In the high glucose medium, the KSC proliferation ability were decreased. The gene expression of fibrosis promote factors after KSC cultured in the high glucose were increased.