糖基化终产物对小鼠胚胎成纤维细胞结缔组织生长因子表达的影响

目的：探讨糖基化终产物（AGEs）对NIH Swiss小鼠胚胎成纤维细胞（NIH/3T3）中结缔组织生长因子（CTGF）基因表达的影响，并观察氨基胍（AG）、葛根素（Pue）对NIH/3T3中CTGF mRNA表达的干预作用。方法：葡萄糖和牛血清白蛋白（BSA）共同孵育制备AGEs，运用荧光法测定AGEs水平。用不同浓度葡萄糖（20、50和80 mmol•L^-1）制备的AGEs培养NIH/3T3细胞24 h；用50 mmol•L^-1葡萄糖制备的AGEs培养NIH/3T3细胞0、6、12、24和48 h。观察AG和不同浓度Pue（0.25、0.5、1.0和1.5g•L^-1）的干预作用。应用RT-PCR检测NIH/3T3中CTGF mRNA的表达。结果：与BSA对照组比较，用20、50和80 mmol•L^-1葡萄糖制备的AGEs培养24 h后，NIH/3T3中CTGF mRNA表达增高（P<0.01），以50 mmol•L^-1最明显。与0 h比较，同一浓度葡萄糖制备的AGEs培养6、12、24及48 h后，NIH/3T3中CTGF mRNA的表达增高（P<0.01），以24 h最明显。与50 mmol•L^-1葡萄糖制备的AGEs培养24 h比较，AG和不同浓度的Pue干预后NIH/3T3中CTGF mRNA的表达明显降低（P<0.01）。结论：AGEs能增强NIH/3T3中CTGF基因表达，且与孵育AGEs的葡萄糖浓度和作用时间有关，提示AGEs可能促进糖尿病并发症纤维化的发展，AG和Pue可抑制AGEs促纤维化的病理过程。

Effect of advanced glycosylation end products on expression of connective tissue growth factor in mouse embryo fibroblasts

Objective To observe the effect of advanced glycosylation end products(AGEs) on gene expression of connective tissue growth factor(CTGF) in NIH Swiss mouse embryo fibroblasts(NIH/3T3),and to assess the intervention actions of aminoguanidine(AG) and puerarin(Pue) on CTGF mRNA expression in NIH/3T3.Methods AGEs were synthesized by coincubation of BSA with glucose.The AGEs content was measured by fluorescence spectroscopy.NIH/3T3 cells were treated with AGEs(prepared with 20,50,80 mmol·L-1 glucose) for 24 h.The NIH/3T3 cells were treated with AGEs(prepared with 50 mmol·L-1 glucose) for 0,6,12,24 and 48 h.The intervention actions of AG and Pue with different concentration(0.25,0.5,1.0 and 1.5 g·L-1) were evaluated.The CTGF mRNA expression in NIH/3T3 was determined by RT-PCR.Results Compared with BSA control,the CTGF mRNA expression levels in NIH/3T3 were increased by treatment with AGEs(prepared with 20,50,80 mmol·L-1 glucose) for 24 h(P<0.01),among them 50 mmol·L-1 glucose was highest.Compared with 0 h,the CTGF mRNA expression levels in NIH/3T3 were increased by treatment with AGEs(prepared with 50 mmol·L-1 glucose) for 6,12,24,48 h(P<0.01),among them 24 h was highest.Compared with AGEs(prepared with 50 mmol·L-1 glucose) for 24 h,AG and Pue with different concentrations significantly decreased the CTGF mRNA expression in NIH/3T3(P<0.01).Conclusion AGEs up-regulates CTGF mRNA expression in NIH/3T3,which is related with glucose concentration and acting time.Based on the above mentioned,AGEs may promote the fibrotic process of diabetic complications.AG and Pue can attenuate the profibrotic effects of AGEs.