The influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats.
To investigate the influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats and explore the mechanisms of this effect. Wistar rats were randomly divided into the following three groups: control, diabetes and fasudil-treatment. All rats were sacrificed after three months of feeding with or without fasudil treatment. Pathological changes to the glomeruli and renal interstitium were studied using Periodic acid-Schiff's staining and Masson staining, respectively. Expression of ROCK1, alpha-SMA, E-cadherin and the distribution of beta-catenin in rat renal cortex were revealed by immunohistochemistry. Changes in the MYPT1 phosphorylation profile and alpha-SMA, E-cadherin and membrane beta-catenin expression were revealed by western blot. Changes in the levels of ROCK1, E-cadherin and total beta-catenin mRNA expression were analyzed by real-time PCR. Fasudil treatment notably attenuates renal interstitial fibrosis in diabetic rats. Compared to the control rats, diabetic rats showed elevated phosphorylation of MYPT1, increased expression of ROCK1 and alpha-SMA, decreased expression of E-cadherin and membrane beta-catenin, and increased expression of ROCK1 and total beta-catenin mRNA, decreased expression of E-cadherin mRNA. Fasudil treatment of diabetic rats resulted in attenuated MYPT1 phosphorylation, decreased ROCK1 and alpha-SMA expression, increased E-cadherin and membrane beta-catenin expression, and reduced ROCK1 and total beta-catenin mRNA expression, increased expression of E-cadherin mRNA. In conclusion, fasudil may reduce the epithelial-mesenchymal transdifferentiation and renal interstitial fibrosis in diabetic rats through a mechanism by which ROCK activity is inhibited, which further facilitates the recovery of the cell-cell adhesions among renal tubular epithelial cells and adhesion complex formation.
Wu
Ganlin
G
Nephrology Department, Renmin Hospital, Wuhan University, China.
Tu
Yafang
Y
Jia
Ruhan
R
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D000117:Acetylglucosamine / Q000652:urine; D000199:Actins / Q000378:metabolism; D000818:Animals; D001786:Blood Glucose / Q000187:drug effects; D015820:Cadherins / Q000378:metabolism; D054338:Cell Transdifferentiation / Q000187:drug effects*; D003404:Creatinine / Q000097:blood; D003921:Diabetes Mellitus, Experimental / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D005355:Fibrosis / Q000188:drug therapy; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007684:Kidney Tubules / Q000187:drug effects; D008297:Male; D054645:Protein Phosphatase 1 / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects; D051176:beta Catenin / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000117:Acetylglucosamine; D000199:Actins; D001786:Blood Glucose; D015820:Cadherins; D003404:Creatinine; D054645:Protein Phosphatase 1; D051176:beta Catenin; D054460:rho-Associated Kinases; C498862:Acta2 protein, rat; C499850:Ppp1r12a protein, rat; C049347:fasudil