Effects Of Spironolactone On Albumin Induced Transdifferentiation Of Human Renal Tubular Epithelial Cell

Objective:To discuss the effects of spironolactone on albumin induced transdifferentiationof human kidney proximal renal tubular epithelial cells(HK-2), and whether theprocess through TGF-β1/Smads signaling pathways.Methods:After be covered about80%~85%and synchronized24hours, HK-2wererandomly divided into the following seven groups in vitro：Group A (withoutstimulus)、Group B (10-7mol/L SPI)、Group C (5mg/ml Alb)、Group D (5mg/mlAlb+10-8mol/l SPI)、Group D (5mg/ml Alb+10-7mol/l SPI)、Group D (5mg/ml Alb+10-6mol/l SPI)、Group G (10umol/l TGF-β1antagonist+30min later adding5mg/ml Alb).Using RT-PCR to detect TGF-β1mRNA and ILKmRNA expression;Immunofluorescence detection in E-cadherin andα-SMA expression; ELISA assayfor detection of FN levels.Results:RT-PCR detection show：In group C TGF beta1mRNA and ILK mRNAexpression was significantly higher than other groups (P <0.01)；In group D, groupE, group F TGF beta1mRNA and ILK mRNA expression gradually declined,between the two groups had significant difference (P <0.05)；Compared with groupC, group G’s TGF beta1mRNA expression had no significant difference (P>0.05),while the expression of ILK mRNA decreased significantly (P <0.01).Cell immunefluorescence detection show that：Group C alpha SMA protein expression wassignificantly higher than other groups (P <0.01), while the expression of E-cadherindecreased obviously (P <0.01)；In group D, group E, group F alpha SMA proteinexpression decreased, E-cadherin protein expression increased gradually (P <0.05)；Compared with group C, group G E-cadherin and alpha SMA protein expression alsohad significant difference (P<0.05).ELISA detection show that：Group C FN proteinlevels in culture medium supernatant higher than other groups (P <0.01)；In group D, group E, group F,the FN protein levels decreased gradually (P <0.05)；Comparedwith group C, group G FN protein level had significant difference (P<0.05).Conclusions:A certain concentration of albumin can be induced epithelial-mesenchymaltransition in vitro;Spironolactone inhibits this process in a dose-dependent manner;TGF-β1/Smads signaling pathway may be play an important role in this process.