1-脱氧野尻霉素对高糖培养小鼠系膜细胞增殖及细胞外基质合成作用研究

Effects of 1- deoxynojirimycin on the proliferation and extracellular matrix synthesis of mouse mesangial cells cultured in high glucose

目的观察1-脱氧野尻霉素(DNJ)对高糖培养小鼠系膜细胞增殖、合成细胞外基质(ECM)的影响。方法体外培养小鼠系膜细胞,用MTT法检测细胞增殖,用ELISA法测定细胞上清液中Ⅳ型胶原、小鼠纤维连接蛋白(FN)含量,用半定量RT-PCR法检测TGF-β_1mRNA、PPARγmRNA表达。结果 (1)高糖组(HG)48 h促细胞增殖(P<0.05);HG 72 h、96 h均抑制细胞增殖(P<0.05)。甘露醇组(MG)与正常糖组(NG)比较差异无统计学意义(P>0.05)。(2)DNJ抑制高糖培养细胞增殖作用呈剂量和时间依赖性。选定HG+5 mmol/L DNJ作用48 h进一步实验。(3)高糖组Ⅳ型胶原、FN合成、TGF-β_1mRNA、PPARr mRNA表达均显著升高(P<0.05或P<0.01)。NG与MG相比差异无统计学意义(P>0.05)。(4)HG+DNJ 5 mmol/L组显著抑制高糖组Ⅳ型胶原、FN合成、TGF-β_1mRNA表达升高(P<0.05),对PPARγmRNA表达升高无影响。结论 DNJ可抑制高糖培养系膜细胞增殖及ECM合成增多,其机制与DNJ降低高糖培养系膜细胞TGFβ_1mRNA表达升高有关。

Objective To investigate the effects of 1- deoxynojirimycin（ DNJ） on the proliferation and the synthesis of extracellular matrix（ ECM） in mouse mesangial cells cultured in high glucose.Methods The mouse mesangial cell line was cultured. The proliferation of the cells was detected by MTT assay. The concentration of type IV collagen（ Col IV） and fibronectin（ FN） was examined with ELISA in cultured medium of mouse mesangial cells. The expression of the mRNA of TGF- β1and PPARγwas detected by RT- PCR. The expression of the protein of TGF- β1 was detected by Western Blot. Results（ 1） High glucose significantly stimulated the proliferation of the cells treated for 48 h as compared to normal glucose group（ NG） and mannitol group（ MG）（ P 〈 0. 05）. Compared with NG and MG,the remarkable inhibition effect was observed by high glucose for 72 h and 96 h（ P 〈 0. 05）. There was no significant difference between mannitol group（ MG） and NG（ P 〉 0. 05）.（ 2） DNJ significantly inhibited the proliferation of mouse mesangial cells cultured in high glucose. The effect of DNJ was time and dosedependent. The further research was made in the cells treated with high glucose ＋ 5 mmol / L DNJ for 48 h.（ 3） High glucose increased the expression level of TGF- β1mRNA and PPARγ mRNA and the synthesis of Col IV and FN（ P 〈 0. 05 或 P 〈 0. 01）. There was no significant difference between MG and NG（ P 〉0. 05）.（ 4） DNJ could decrease the expression level of TGF- β1mRNA and the synthesis of Col IV and FN of cells cultured in high glucose（ P 〈 0. 05）,while DNJ had no effect on increased expression level of PPARγ mRNA of cells cultured in high glucose. Conclusion DNJ inhibits proliferation and ECM synthesis of mesangial cells cultured in high glucose,which may be mediated by downregulating the expression of TGF- β1mRNA.