Augmenter Of Liver Regeneration On Renal Tubular Epithelial Cell Transdifferentiation Of Rat Study On The Effect

Objective: To express rALR in GS115 with constructed pichia pastoris expression plasmid of rat augmenter of liver regeneration (rALR) and purify it by ultrafiltration and identify its bioactivity in vitro, for studying its biological functions.Methods: The recombinant plasmid pPIC9K-rALR was transformed into GS115 by electroporation. The rALR was expressed in GS115 under the induction of methanol and purified with ultrafiltration. The purified product was identified by 15% SDS-PAGE and Western blot. The effects of rALR on the proliferations of QGY were evaluated by 3H-TdR methods in vitro.Results: The rALR was efficiently expressed as a secretive protein in GS115. Its molecular weight(1.5×104 dalton) was in correspondance with theoretic values. Single band of rALR was observed in SDS-PAGE electrophoresis and in western blot after ultrafiltration. The rALR promoted the proliferation of QGY by a dose-dependent way in vitro. Conclusion: The rALR was efficiently expressed in GS115 and successfully purified by ultrafiltration. The proliferation of QGY was stimulated by rALR in vitro with a dose-dependent way.Objective: To investigate the effect of recombination rat augmenter of liver regeneration(rrALR) on TGF-β1-induced renal tubular epithelial cells transdifferentiation(TEMT). The cells were cultured for 48 hours. Morphological change of cellsMethods: The cells of normal rat kidney tubular epithelial cell line(NRK-52E) were divided into four groups, normal control cells, rrALR treated cells, TGF-β1 treated cells, TGF-β1 plus rrALR treated cells. was observed and expressions ofα-SMA, CTGF, E-cadherin protein and mRNA were analysed. Results: The results showed that TGF-β1 induced cell morphological change of NRK-52E as the development of an elongated and fusiforill appearance, and up-regulated protein and mRNA expressions ofα-SMA, CTGF and down-regulated E-cadherin expression. Transformation of tubular epithelial cells was inhibited in TGF-β1 plus rrALR group as compared with TGF-β1 treated group. The protein and mRNA expression of E-cadherin was increased and levels ofα-SMA and CTGF were decreased in cells exposed TGF-β1 plus rrALR medium.Conclusion: 1.There was no transdifferentiating effect when NRK52E cells cultured with rALR alone. 2. rrALR may inhibit transdifferentiation of NRK52E cells into myofibroblast-like cells and CTGF protein and mRNA expression induced by TGF-β1.