他克莫司对肾小管上皮细胞转分化早期CTGF、VEGF表达的影响

Effects of Tacrolimus on transdifferentiation and CTGF,VEGF early expression of human renal tubular epithelial cells

目的探讨不同剂量的他克莫司(FK506)对肾小管上皮细胞(HKC)转分化的作用及其对早期结缔组织生长因子(CTGF)、血管内皮生长因子(VEGF)表达的影响。方法以体外培养的HKC为研究对象,分4组:(1)对照组;(2)转化生长因子-β1(TGF-β1)(8ng/mL)组;(3)FK506(10、30、60ng/mL)组;(4)TGF-β1(8ng/mL)加FK506(10、30、60ng/mL)组。应用形态学、免疫组化技术、和半定量反转录PCR(RT-PCR)观察FK506对TGF-β1诱导的HKC转分化的作用及对α-SMA、CTGF、VEGF表达的影响。结果与对照组比较,48h后TGF-β1组HKC细胞呈现出长梭形外观,α-平滑肌肌动蛋白(α-SMA)、CTGF、VEGF的表达明显增多(P<0.01),FK506组α-SMA、VEGF及CTGF的表达差异无统计学意义(P>0.05)。TGF-β1加FK506组比TGF-β1组α-SMA、VEGF及CTGF随FK506的浓度增加表达明显降低(P<0.05)。结论 FK506对TGFβ1诱导的肾小管上皮细胞转分化早期可能通过下调VEGF、CTGF的表达起抑制作用。

Objective To explore different doses of tacrolimus（FK506） to renal tubular epithelial cells（HKC） transdifferentia tion and its early action on connective tissue growth factor（CTGF）,vascular endothelialgrowth factor （VEGF） expressing change. Methods The human kidney cells（HKC） were cultured for 48 hours in different conditions： （1）Serum free as control, （2）Treated with TGF-β1（8 ng/mL） ;（3）Treated with FK506 at different concentration（10,30,60 ng/mL） ; （4）Treated with FK506 at different concentration（10,30,60 ng/mL） plus TGF-β1 （ng/mL）. The expression of α-SMA was assessed with RT-PCR,The expression of CTGF,VEGF was assessed with RT-PCR and immunohistochemistry after 48 hours. Results Compared with the control group, The expression of CTGF, VEGF and α-SMA mRNA was markedly increased in HCK cultured with TGF-βI group（P〈0.05）, which were suppressed significantly in HKC cultured with TGF-β1 plus FK506 group（P〈0.05）. Conclusion The inhibition of FK506 on transdifferentiation of tubular epithelial ceils may be related to the down-regulation of VEGF,CTGF.