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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 effect of Smad2 antisense oligonucleotide (ODN) on the secretion of fibronectin (FN) and type Ⅳ collagen (Ⅳ) in mesangial cells cultured in high glucose and to explore the role of Smad2 in glomerulosclerosis A new method of glomerulosclerosis. Methods The sense, antisense and random ODNs of Smad2 start codon were designed and synthesized. ODN was transiently transfected into rat mesangial cells by lipofectamine and cultured in high glucose. To observe the expression of Smad2 mRNA and protein and its effect on the secretion of extracellular matrix (ECM) of mesangial cells. Results The Smad2 antisense ODN transfected rat mesangial cells showed a decrease in Smad2 mRNA and protein expression compared with the high glucose group, the normal group and the randomized group (P <0 郾 05) Group compared with no significant (P> 0.05). The content of FN (ng / ml) in high glucose group, antisense group, sense group and randomized supernatant were 84.19 ± 6.81,57.65 ± 5.05,78.72 ± 9.13 and 77.13 ± 7.36, respectively. Col Ⅳ content (ng / ml) Respectively, 55.27 ± 4.75, 22.31 ± 3.24, 46.23 ± 2.02, 45.92 ± 1.21. Antisense ODN group FN, ColⅣ content were reduced, and high glucose, sense, random ODN group compared with the difference was statistically significant (P <0.05). Conclusion Antisense Smad2ODN can inhibit the up-regulation of Smad2FN and Col in mesangial cells induced by high glucose. The results provide a new way of thinking and treatment for delaying renal fibrosis.