摘要目的 了解体外高糖刺激对肾小管上皮细胞(HK-2)脂联素受体(adipoR)表达的影响.方法 将HK-2细胞在含10%胎牛血清的低糖DMEM培养液中培养至贴壁且80%融合时,无血清培养24 h,改用含葡萄糖1、2,4、6、8 mg/ml的培养液继续培养48 h,RT-PCR及westernblot检测ad-ipoR的表达.将HK-2细胞分别在含10%胎牛血清的高糖(4 mg/ml)、低糖(1 ms/ml)DMEM培养液中培养0、12、24、48、72、96 h后提取总RNA,RT-PCR检测adipoR mRNA的表达.结果 adipoR1/R2基因在HK-2细胞上均有表达,且adipoR1(0.63±0.12)基因表达量是adipoR2(0.16±0.03)的3.9倍,差异有统计学意义(P<0.01).各不同浓度葡萄糖及作用时间对HK-2细胞adipoR的基因表达无明显影响(P>0.05).westernblot检测到HK-2细胞上adipoR1的蛋白表达,其表达量不受葡萄糖浓度的影响(P>0.05).结论 adipoR1/R2基因在HK-2细胞上均有表达,且以adipoR1为主,提示adipoR1在肾脏疾病中具有更重要的意义.HK-2细胞adipoR的表达不受高糖的影响.

abstractsObjective To investigate the effect of in vitro high glucose stimulation on the expression of adiponectin receptor (adipoR) in human kidney proximal tubular cells.Methods The HK-2 cells were cultured in the low glucose DMEM culture medium containing 10% fetal bovine serum until the cells were adherent and 80% confluence. After cultured in the serum-free DMEM for 24 hours, these cells were stimulated with glucose-containing 1mg/ml, 2mg/ml, 4mg / ml, 6mg/ml, 8mg/ml serum-free DMEM for 48 hours. Then RT-PCR and western blot were used to analyze adipoR ( R1, R2) expression levels. The HK-2 cells were cultured respectively in high glucose (4mg/ml) , low glucose (1mg/ml) DMEM culture medium containing 10% fetal bovine serum to cultivate 0h, 12h, 24h, 48h, 72h, 96h, then RT - PCR was applied to analyze adipoR (R1, R2) mRNA expression levels semi-quantitatively. Results Two kinds of adiponectin receptor gene were both expressed in HK-2 cells, and the quantity of gene expression of adipoR1 (0. 63 ±0. 12) was 3. 9 times to adipoR2 (0. 16 ±0.03) , the difference was statistically significant ( P<0. 01). The different concentrations of glucose and different time of high glucose on HK-2 cells had no significant effect ( P>0. 05 ) on adipoR gene expression. Expression of adipoR 1 protein in HK-2 cells was detected by western blot, and it was not affected by glucose concentration ( P>0. 05).Conclusion adi-poR1 and adipoR2 gene were both expressed in HK-2 cells, and the adipoR1 was the major one, which suggested that adipoR1 played a more significant role in kidney disease. The expression of adipoRl/R2 of HK-2 cells was not affected by high glucose concentration.