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目的:观察体外转染结缔组织生长因子(CTGF)反义寡核苷酸(ASODN)对高糖培养的人肾小球系膜细胞(HMC)细胞外基质表达的影响。方法:体外培养HMC,予以高糖(30mmol/L)和正常糖(5mmol/L)培养液培养,高糖培养后应用脂质体Lipofectamine2000转染CTGFASODN,在不同时间收集培养液上清。用ELISA检测培养液上清中的纤连蛋白(FN)和层黏连蛋白(LN)含量。结果:高糖呈时间依赖促进体外培养的HMC表达FN和LN,而转染CTGFASODN可抑制高糖所致的FN和LN分泌增加。结论:高糖培养可促进HMC细胞外基质的表达,转染CTGFASODN可抑制上述高糖的作用。故调控HMC的CTGF表达水平可能成为防治糖尿病肾病(DN)、特别是早期病变的有效途径之一。
OBJECTIVE: To observe the effects of antisense oligonucleotide (CTGF) antisense oligonucleotide (ASODN) transfected in vitro on the expression of extracellular matrix in human mesangial cells (HMC) cultured in high glucose. Methods: HMC was cultured in vitro and cultured in high glucose (30 mmol / L) and normal glucose (5 mmol / L). Lipofectamine 2000 was used to transfect CTGFASODN in high glucose and supernatants were collected at different times. The content of fibronectin (FN) and laminin (LN) in the culture supernatant was detected by ELISA. Results: High glucose showed a time-dependent effect on FN and LN expression in HMC cultured in vitro. Transfection of CTGFASODN inhibited the increase of FN and LN secretion induced by high glucose. Conclusion: High glucose can promote the expression of extracellular matrix of HMC. Transfection of CTGFASODN can inhibit the above high glucose. Therefore, the regulation of CTGF expression in HMC may be one of the effective ways to prevent and treat diabetic nephropathy (DN), especially early lesions.