摘要目的 体外实验研究脂联素对高糖刺激下的大鼠近端肾小管上皮细胞(NRK-52E)单核趋化蛋白-1(MCP-1)mRNA及蛋白表达的影响.方法 用含不同浓度葡萄糖的DMEM培养基体外培养NRK-52E细胞,分5组(每组4个样本,此实验重复4次):A组:含5 mmol/L葡萄糖培养基对照组;B组:含30 mmol/L葡萄糖培养基组;C组:含30 nmol/L葡萄糖培养基+1 mg/L脂联素组;D组:含30 mmol/L葡萄糖培养基+5 mg/L脂联素组;E组:含30 mmol/L葡萄糖培养基+10 mg/L脂联素组.以逆转录-聚合酶链反应(RT-PCR)、Western blot法榆测比较各组细胞MCP-1 mRNA及蛋白表达的变化.组间比较采用t检验,多组间比较采用方差分析.结果 A组细胞MCP-1 mRNA表达量为0.247±0.005,B组为0.691±0.009,显著高于A组(t=72.03,P＜0.01);C组为0.425±0.013,显著高于A组(t=46.31,P＜0.05);D组为0.307±0.012,与A组相近(t=73.24,P＞0.05);E组为0.253±0.011,与A组无差异.不同浓度脂联素各组间比较差异亦具有统计学意义(F=37.15,P＜0.05).A组MCP-1蛋白表达量为10.25±0.03,B组为58.47±0.02,显著高于A组(t=35.21,P＜0.01);C组为35.86±0.05,较B组显著下降(t=48.26.21,P＜0.05);D组为25.63±0.06,较B组显著下降(t=32.34,P＜0.01);E组为21.53±0.03,较B组显著下降(t=42.26,P＜0.05),但高于A组(t=64.28,P＜0.01).不同浓度脂联素各组间比较差异亦具有统计学意义(F=53.15,P＜0.05).结论 脂联素可呈剂量依赖性地抑制高糖环境下大鼠近端肾小管上皮细胞MCP-1 mRNA及蛋白的高表达.

abstractsObjective To observe the effect of adiponectin on the secretion of monocyte chemotactic protein( MCP-1 ) in rat proximal renal tubular epithelial cell(NRK-52E) in vitro. Methods Using DMEM medium with different concentrations of glucose and adiponectin to culture rat proximal tubular epithelial cells in vitro in five groups(four samples in each group,the experiment were repeated for 4 times): group A:DMEM medium with 5 mmol/L of glucose as control group; group B: DMEM medium with 30 mmol/L of glucose; group C: DMEM medium with 30 mmol/L of glucose and 1 mg/L of recombinant adiponectin;group D: DMEM medium with 30 mmol/L of glucose medium and 5 mg/L of recombinant adiponectin; group E: DMEM medium with 30 mmol/L of glucose medium and 10 mg/L of recombinant adiponectin. The reverse transcription time-polymerase chain reaction(RT-PCR) and Western blot methods were used to detect the expression of MCP-1 mRNA and protein in the 5 groups. The results were compared among the 5 groups.The t test was used when compared between two groups and amalysis of variance was used in the comparison among multi-groups. Results The expression of MCP-1 mRNA in group A was 0. 247 ± 0. 005,it was 0. 691 ±0. 009 in group B and was significantly higher than that in group A( t =72.03,P ＜0. 01 ); it was 0.425 ±0.013 in group C and was significantly higher than that in group A(t =46.31,P ＜0.05); it was 0. 307 ±0. 012 in group D and was higher than that in group A with no significant differences( t= 73.24,P ＞ 0. 05 ); it was 0. 253 ± 0. 011 in group E and was similar with group A. There was significant differences in expression of MCP-1 mRNA among the groups with different concentrations of adiponectin( F = 37. 15 ,P ＜0. 05 ). The expression of MCP-1 protein in group A was 10. 25 ±0. 03,it was 58.47 -0. 02 in group B and was significantly higher than that in group A ( t = 35.21 ,P ＜0. 01 ); it was 35.86 ±0. 05 in group C and was significantly lower than that in group B ( t = 48.26. 21,P ＜ 0. 05 ); it was 25.63 ± 0. 06 in group D and was significantly lower than that in group B ( t = 32. 34,P ＜ 0. 01 ); it was 21.53 ± 0. 03 in group E and was significantly lower than that in group B ( F= 42. 26,P ＜ 0. 05 ) and significantly higher than that in group A (t-=64. 28,P ＜0. 01 ). There was significant differences in expression of MCP-1 protein among the groups with different concentrations of adiponectin( F= 53. 15,P ＜ 0. 05 ). Conclusion Adiponectin could inhibit the highly expressed MCP-1 mRNA and protein induced by high glucose in rat proximal tubular epithelial cells in vitro in a dose-dependent manner.