| Objective: To observe the effect of Transforming Growth Factor-β1(TGF-β1) on the expression of tissue Transglutaminase (tTG) in culturedhuman renal interstitial fibroblasts. Methods: 1.Human renal fibroblasts were successfully cultured from isolatedpapilla of human kidney and identified by morphologic examination,immunocytochemistry staining and cultured in D-valine selective MEMmedium. 2.The fibroblasts were incubated with different concentrations ofTGF- β 1 (1ng/ml, 2ng/ml, 5ng/ml) for 24 hours respectively, tTGexpression was determined by RT-PCR and Western blot. 3.The fibroblasts were cultured with TGF-β1 for different time(8hr, 24hr, 36hr) at the concentration of 2ng/ml, tTG expression wasdetermined by RT-PCR and Western blot. 4.The fibroblasts were incubated with TGF-β1(2ng/ml). Theexpression of tTG was detected by immunocytochemistry staining. Results: 1.Human renal interstitial fibroblasts were showed as shuttle-shapedcells. They had positive staining with anti-vimentin antibody. They couldnot grow in the selective medium and died partly in a week. 2.There was basic expression of tTG mRNA in normal human renalinterstitial fibroblasts, and TGF-β1 was found to upregulate the mRNAexpression of tTG significantly. 3.There was low level expression of tTG protein in normal humanrenal interstitial fibroblasts. TGF-β1 can up-regulate the expression of tTGsignificantly ,and the increase of tTG is time-dependent anddose-dependent . Conclusions: There was basic expression of tTG in the normal human renalinterstitial fibroblasts. TGF-β1 can up-regulate the expression of mRNAand protein of tTG. The interaction between TGF-β1 and tTG may beinvolved in the renal interstitial fibrosis . |
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