摘要
目的观察Smad2反义寡核苷酸(ODN)对高糖培养的系膜细胞分泌纤连蛋白(FN)、Ⅳ型胶原(ColⅣ)的影响,探讨Smad2在肾小球硬化中的作用,寻找延缓肾小球硬化的新方法。方法设计、合成Smad2起始密码子部位的正义、反义和随机ODN,用脂质体法将ODN瞬时转染大鼠系膜细胞,并进行高糖培养。观察Smad2mRNA及蛋白的表达及其对系膜细胞细胞外基质(ECM)分泌的影响。结果转染Smad2反ODN义的大鼠系膜细胞,其Smad2mRNA及蛋白表达均降低,与高糖组、正义组和随机组相比差异有统计学意义(P<0郾05),与正常对照组相比无统计学意义(P>0.05)。高糖组、反义组、正义组和随机组培养上清液中FN含量(ng/ml)分别为84.19±6.81、57.65±5.05、78.72±9.13、77.13±7.36;ColⅣ含量(ng/ml)分别为55.27±4.75、22.31±3.24、46.23±2.02、45.92±1.21。反义ODN组FN、ColⅣ的含量均减少,与高糖、正义、随机ODN组相比差异有统计学意义(P<0.05)。结论反义Smad2ODN可抑制高糖刺激引起的系膜细胞Smad2FN和Col的表达上调,该研究结果为延缓肾脏纤维化提供了新的思路和治疗途径。
Objective To investigate the effects of antisense Smad2 oligodeoxynudeotides(ODN) on fibronetin(FN) and collagen Ⅳ(ColⅣ) secretion of rat mesangial cells cultured with high glucose, explore the action of Smad2 in the glomerulosclerosis and to find a new method to retard the progress of glomerular fibrosis. Methods 20-mer antisense, sense and random ODNs were designed and synthesized that were phosphorothioate modified to increase stability. The antisense ODN encompassed the ATG of the rat Smad2 gene. ODN was tranferred transiently into rat mesangial cells through liposome. Rat cells were treated with high glucose. mRNA and protein of Smad2 were detected by RT-PCR and cytochemistry. FN and ColⅣ were examined by ELISA. Results Antisense ODN significantly decreased mRNA and protein expression of Smad2 in rat mesangial cells treated with high glucose(P < 0.05). FN concentrations in the culture medium of high glucose group, antisense group, sense group and random group were (84.19±6.81) ng/ml, (57.65±5.05) ng/ml, (78.72±9.13) ng/ml and (77.13±7.36) ng/ml respectively. ColⅣ concentrations were (55.27±4.75) ng/ml, (22.31±3.24) ng/ml, (46.23±2.02) ng/ml and (45.92±1.21) ng/ml respectively. Antisense ODN significantly decreased the secretion of FN and ColⅣ in the culture medium (P < 0.05). Conclusion Antisense Smad2 ODN can reduce the FN and Col IV secretion in rat mesangial cells cultured with high glucose.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2005年第2期103-107,共5页
Chinese Journal of Nephrology
基金
上海市科学委员会博士后资助计划科学基金(2001-1178)
