摘要
目的 :探讨己糖胺通路 (HBP)在高糖引起系膜细胞功能改变中的作用。 方法 :利用体外培养的人肾小球系膜细胞 ,流式细胞术分析细胞体积、细胞周期和纤维连接蛋白 (FN)的含量 ,以3H 胸腺嘧啶的掺入反映细胞的增生。采用夹心ELISA法和RT PCR检测转化生长因子 β1(TGF β1)蛋白质和mRNA的表达。采用比色法测定HBP限速酶—谷氨酰胺 :6 磷酸果糖转氨酶 (GFAT)的活性。 结果 :HBP激活剂 葡萄糖胺能够诱导系膜细胞肥大、增生抑制和FN合成增多 ,并增加TGF β1的表达。高糖能增加GFAT的活性 ,高糖促进TGF β1表达的作用能够被GFAT的抑制剂—重氮丝氨酸拮抗。 结论 :HBP的过度活化 ,可能通过上调TGF β1的表达 ,从而介导了高糖对系膜细胞的损伤。
To investigate the role of hexosamine biosynthesis pathway(HBP)in high glucose-induced phenotypic changes in mesangial cells in vitro. METHODOLOGY Cultured human mesangial cells were used in this in vitro study.Cell volume,cell cycle and fibronectin production were analyzed by flow cytometry.Cell proliferation was determined by 3H-Tdr incorporation assay.The expression of transforming growth factor β1(TGF-β1)protein and mRNA were determined by sandwich ELISA and RT-PCR respectively.The activity of glutamine:fructose-6-phosphate aminotransferase(GFAT),which is the key enzyme of HBP,was measured by spectrophotometry method. RESULTS Glucosamine,an activator of the activity of HBP,enhanced the expression of TGF-β1 and the production of matrix component fibronectin,and inhibited cell proliferation in a time-and dose-dependent manner.High glucose increased the activity of GFAT.Inhibition of GFAT by the substrate analogue azaserine prevented the high glucose induced increase in TGF-β1 expression. CONCLUSION The results suggest that high glucose induced mesangial cell TGF-β1 synthesis is mediated by upregulated HBP.Excessive flux of glucose through HBP may play an important role in the pathogenesis of diabetic nephropathy.
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
2000年第4期303-310,共8页
Chinese Journal of Nephrology,Dialysis & Transplantation
基金
国家自然科学基金!(39870 2 88)资助项目
关键词
糖尿病肾病
转化生长因子Β1
已糖胺通路
transforming growth factor β1 diabetic nephropathy hexosamine biosynthesis pathway
