高糖对大鼠结肠平滑肌细胞表达内源性胰岛素样生长因子1的影响  

High glucose down-regulates the expression of endogenous insulin-like growth factor-1 in rat colonic smooth muscle cells

作　　者：徐新寓[1] 王云[1] 杨微微[1] 林琳[1] 

机构地区：[1]南京医科大学第一附属医院消化科,江苏省南京市210029

出　　处：《世界华人消化杂志》2012年第12期998-1003,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30971354;江苏省国际科技合作计划基金资助项目;No.BZ2011044;江苏省研究生教育创新工程基金资助项目;No.CXZZ11_0704~~

摘　　要：目的:探讨高糖对大鼠结肠平滑肌细胞(SMCs)表达内源性胰岛素样生长因子1(IGF-1)的影响.方法:酶解法分离培养SD大鼠结肠SMCs,α-actin免疫荧光鉴定,然后将大鼠结肠SMCs随机给予葡萄糖不同浓度(5.5mmol/L和25mmol/L)组及甘露醇对照组(5.5mmol/L葡萄糖+19.5mmol/L甘露醇)刺激,CCK8实验检测SMCs增殖情况;流式细胞术检测SMCs细胞周期;ELISA检测培养液上清中IGF-1的含量;Western blot、Real-time PCR法检测SMCs合成内源性IGF-1的表达变化.结果:高糖(25mmol/L)抑制大鼠结肠SMCs的增殖,在24h与正常糖浓度间差异最大(0.494±0.003vs0.597±0.044,P<0.05);高糖使约90%的结肠SMCs停滞在G1期(90.850%±0.706%vs55.202%±3.807%,P<0.05),进入S期的SMCs明显减少(3.622%±0.156%vs30.780%±3.808%,P<0.05);高糖环境中,结肠SMCs合成分泌的IGF-1减少(208.000ng/L±31.443ng/Lvs265.750ng/L±26.538ng/L,P<0.05),SMCs表达内源性的IGF-1 mRNA和蛋白也均减少(2.037±0.196vs2.257±0.273;0.247±0.045vs0.906±0.103,P<0.05).结论:高糖抑制大鼠结肠SMCs增殖,使SMCs内源性IGF-1表达减少.AIM：To investigate the effect of high glucose on the expression of endogenous insulin-like growth factor-1（IGF-1）in rat colonic smooth muscle cells（SMCs）.METHODS：Rat colonic SMCs were separated,cultured,identified by immunofluorescence staining of？-actin,and divided into three groups：normal glucose group（5.5 mmol/L glucose）,mannitol control group（5.5 mmol/L glucose plus 19.5 mmol/L mannitol）and high glucose group（25 mmol/L glucose）.After treatment,cell proliferation was determined using Cell Counting Kit-8,and cell cycle analysis was performed by flow cytometry.ELISA was designed to measure the content of IGF-I in SMCs culture supernatants.Real-time quantitative-PCR and Western blotting were performed to analyze the mRNA and protein expression of IGF-1 in SMCs.RESULTS：Compared to the normal glucose group,treatment with high glucose significantly inhibited the proliferation of rat colonic SMCs（0.494±0.0030 vs 0.597±0.044,P0.05）,resulted in cell accumulation in the G1 phase（90.850%±0.706%vs 55.202%±3.807%,P0.05） and a significant decrease in the percentage of cells in the S phase（3.622%±0.156%vs 30.780% ±3.808%,P0.05）,and decreased the content of IGF-I in SMCs culture supernatants（208.000 ng/L±31.443 ng/L vs 265.750 ng/L±26.538 ng/L,P0.05）and the expression of IGF-I mRNA and protein（2.037±0.196 vs 2.257±0.273;0.247± 0.045 vs 0.906±0.103,both P0.05）.However,there were no significant differences in the above parameters between the normal glucose group and mannitol control group.CONCLUSION：High glucose inhibits the proliferation of rat colonic SMCs and decreases the expression of endogenous IGF-1 in SMCs.

关 键 词：高糖 结肠平滑肌细胞 细胞周期 胰岛素样生长因子1 

分 类 号：R587.1[医药卫生—内分泌]