高糖对人肾近端小管细胞TGF-β mRNA表达的影响

Effect of high concentration of glucose on the expression of TGF-β mRNA and its relationship with the effect of angiotensin Ⅱ in human proximal tubular cells

目的 观察高糖对人近端肾小管细胞 (HPTC)转化生化因子 (TGF β)mRNA表达的影响以及血管紧张素Ⅱ (ATⅡ )受体 1拮抗剂Irbesartan(Irb)对其干预作用。方法 无血清培养HPTC后 ,(1)用不同浓度葡萄糖 (5 5、2 2 2、30、4 0mmol/L)培养 4 8h ,(2 )用高糖 (2 2 2mmol/L)培养 6、12、2 4、4 8、72h ,(3)高糖培养液中加入不同浓度的Irb(10 -3 、10 -5、10 -7mol/L)作用 4 8h ;采用逆转录聚合酶链反应 (RT PCR)技术测定HPTC表达TGF βmRNA的水平变化。 结果 高糖能促进HPTCTGF βmRNA的表达 ,且呈浓度和时间依赖性。Irb能部分降低高糖诱导HPTCTGF βmRNA的表达 ,且呈剂量依赖性。结论 高糖可刺激肾小管细胞表达TGF β增加 。

Objective To observe the effect of high concentration of glucose on transforming growth factor-β(TGF-β)mRNA expression and the influence of Irbesartan in human proximal tubular cells(HPTC).Methods After 24 h of serum depletion,cells were incubated in different concentration of D-glucose(5 5,22 2,30,40 mmol/L)for 48 hours and were incubated in 22 2 mmol/L D-glucose for times up to 72 hours.Growth-arrested HPTC was exposed to serum-free medium containing 22 2 mmol/L D-glucose and different concentrations of Irbesartan(Irb)(10 -3,10 -5,10 -7 mol/L)for 48 hours.Total cellular RNA was isolated from cultured HPTC by TRIzol method,and expression of TGF-βmRNA was assessed by RT-PCR,which was performed by using access RT-PCR kit(Promega).Results D-glucose stimulated the expression of TGF-β mRNA in HPTC in time and dose dependent manner.Irb(10 -3,10 -5,10 -7 mol/L)could dose dependently attenuate the mRNA expression stimulated by D-glucose(22 2 mmol/L)for 48 h.Conclusion Our results suggest that high concentration of glucose may stimulate the expression of TGF-β through activation of the intrarenal rennin angiotensin system.