中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2011年
2期
147-150
,共4页
姚涛%吴小燕%於文丽%高苹%曾真
姚濤%吳小燕%於文麗%高蘋%曾真
요도%오소연%어문려%고평%증진
糖尿病肾病%大鼠%单核细胞趋化蛋白-1%脂联素%近端肾小管上皮细胞
糖尿病腎病%大鼠%單覈細胞趨化蛋白-1%脂聯素%近耑腎小管上皮細胞
당뇨병신병%대서%단핵세포추화단백-1%지련소%근단신소관상피세포
Diabetic nephropathies%Rats%Monocyte chemotactic protein 1%Adiponectin%Proximal renal tubular epithelial cell
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.
目的 體外實驗研究脂聯素對高糖刺激下的大鼠近耑腎小管上皮細胞(NRK-52E)單覈趨化蛋白-1(MCP-1)mRNA及蛋白錶達的影響.方法 用含不同濃度葡萄糖的DMEM培養基體外培養NRK-52E細胞,分5組(每組4箇樣本,此實驗重複4次):A組:含5 mmol/L葡萄糖培養基對照組;B組:含30 mmol/L葡萄糖培養基組;C組:含30 nmol/L葡萄糖培養基+1 mg/L脂聯素組;D組:含30 mmol/L葡萄糖培養基+5 mg/L脂聯素組;E組:含30 mmol/L葡萄糖培養基+10 mg/L脂聯素組.以逆轉錄-聚閤酶鏈反應(RT-PCR)、Western blot法榆測比較各組細胞MCP-1 mRNA及蛋白錶達的變化.組間比較採用t檢驗,多組間比較採用方差分析.結果 A組細胞MCP-1 mRNA錶達量為0.247±0.005,B組為0.691±0.009,顯著高于A組(t=72.03,P＜0.01);C組為0.425±0.013,顯著高于A組(t=46.31,P＜0.05);D組為0.307±0.012,與A組相近(t=73.24,P＞0.05);E組為0.253±0.011,與A組無差異.不同濃度脂聯素各組間比較差異亦具有統計學意義(F=37.15,P＜0.05).A組MCP-1蛋白錶達量為10.25±0.03,B組為58.47±0.02,顯著高于A組(t=35.21,P＜0.01);C組為35.86±0.05,較B組顯著下降(t=48.26.21,P＜0.05);D組為25.63±0.06,較B組顯著下降(t=32.34,P＜0.01);E組為21.53±0.03,較B組顯著下降(t=42.26,P＜0.05),但高于A組(t=64.28,P＜0.01).不同濃度脂聯素各組間比較差異亦具有統計學意義(F=53.15,P＜0.05).結論 脂聯素可呈劑量依賴性地抑製高糖環境下大鼠近耑腎小管上皮細胞MCP-1 mRNA及蛋白的高錶達.
목적 체외실험연구지련소대고당자격하적대서근단신소관상피세포(NRK-52E)단핵추화단백-1(MCP-1)mRNA급단백표체적영향.방법 용함불동농도포도당적DMEM배양기체외배양NRK-52E세포,분5조(매조4개양본,차실험중복4차):A조:함5 mmol/L포도당배양기대조조;B조:함30 mmol/L포도당배양기조;C조:함30 nmol/L포도당배양기+1 mg/L지련소조;D조:함30 mmol/L포도당배양기+5 mg/L지련소조;E조:함30 mmol/L포도당배양기+10 mg/L지련소조.이역전록-취합매련반응(RT-PCR)、Western blot법유측비교각조세포MCP-1 mRNA급단백표체적변화.조간비교채용t검험,다조간비교채용방차분석.결과 A조세포MCP-1 mRNA표체량위0.247±0.005,B조위0.691±0.009,현저고우A조(t=72.03,P＜0.01);C조위0.425±0.013,현저고우A조(t=46.31,P＜0.05);D조위0.307±0.012,여A조상근(t=73.24,P＞0.05);E조위0.253±0.011,여A조무차이.불동농도지련소각조간비교차이역구유통계학의의(F=37.15,P＜0.05).A조MCP-1단백표체량위10.25±0.03,B조위58.47±0.02,현저고우A조(t=35.21,P＜0.01);C조위35.86±0.05,교B조현저하강(t=48.26.21,P＜0.05);D조위25.63±0.06,교B조현저하강(t=32.34,P＜0.01);E조위21.53±0.03,교B조현저하강(t=42.26,P＜0.05),단고우A조(t=64.28,P＜0.01).불동농도지련소각조간비교차이역구유통계학의의(F=53.15,P＜0.05).결론 지련소가정제량의뢰성지억제고당배경하대서근단신소관상피세포MCP-1 mRNA급단백적고표체.
Objective 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.
