Alpha1-Antitrypsin Attenuates Renal Fibrosis by Inhibiting TGF-β1-Induced Epithelial Mesenchymal Transition.
Alpha1-antitrypsin (AAT) exerts its anti-inflammatory effect through regulating the activity of serine proteinases. This study evaluated the inhibitory effects of AAT against the transforming growth factor (TGF)-β1 induced epithelial-to-mesenchymal transition (EMT) in unilateral ureter obstruction (UUO) mice and Madin-Darby canine kidney (MDCK) cells. C57BL/6 mice with induced UUO were injected intraperitoneally with AAT (80 mg/Kg) or vehicle for 7 days. MDCK cells were treated with TGF-β1 (2 ng/mL) for 48 hours to induce EMT, and co-treated with AAT (10 mg/mL) to inhibit the EMT. Masson's trichrome and Sirius red staining was used to estimate the extent of renal fibrosis in UUO mice. The expression of alpha-smooth muscle actin (α-SMA), vimentin, fibronectin, collagen I, and E-cadherin in MDCK cells and kidney tissue were evaluated. Masson's and Sirius red staining revealed that the area of renal fibrosis was significantly smaller in AAT treated UUO group compared with that of UUO and vehicle treated UUO groups. AAT treatment attenuated upregulation of Smad2/3 phosphorylation in UUO mouse model. Co-treatment of MDCK cells with TGF-β1 and AAT significantly attenuated the changes in the expression of α-SMA, vimentin, fibronectin, collagen I, and E-cadherin. AAT also decreased the phosphorylated Smad3 expression and the phosphorylated Smad3/Smad3 ratio in MDCK cells. AAT treatment inhibited EMT induced by TGF-β1 in MDCK cells and attenuated renal fibrosis in the UUO mouse model. The results of this work suggest that AAT could inhibit the process of EMT through the suppression of TGF-β/Smad3 signaling.
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Jang-Hee
JH
Cho
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Hye-Myung
HM
Ryu
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Eun-Joo
EJ
Oh
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Ju-Min
JM
Yook
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Ji-Sun
JS
Ahn
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Hee-Yeon
HY
Jung
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Ji-Young
JY
Choi
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Sun-Hee
SH
Park
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Yong-Lim
YL
Kim
Department of Internal Medicine, Pusan National University Hospital, Busan, Korea.
Ihm Soo
IS
Kwak
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Chan-Duck
CD
Kim
D000199:Actins / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D004285:Dogs; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis; D005455:Fluorescent Antibody Technique; D007668:Kidney / Q000473:pathology*; D061985:Madin Darby Canine Kidney Cells; D008297:Male; D008648:Mesoderm / Q000187:drug effects / Q000378:metabolism; D008810:Mice, Inbred C57BL; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D051785:Smad Proteins / Q000378:metabolism; D013194:Staining and Labeling; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects; D014517:Ureteral Obstruction / Q000188:drug therapy / Q000473:pathology; D014746:Vimentin / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000627:therapeutic use*
C558396:Acta2 protein, mouse; D000199:Actins; D015415:Biomarkers; D015820:Cadherins; D024042:Collagen Type I; D005353:Fibronectins; D012333:RNA, Messenger; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; D014746:Vimentin; D000515:alpha 1-Antitrypsin