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Novel RAS inhibitor 25-O-methylalisol F attenuates epithelial-to-mesenchymal transition and tubulo-interstitial fibrosis by selectively inhibiting TGF-β-mediated Smad3 phosphorylation.
Phytomedicine. 2018 Mar 15; 42:207-218.P

Abstract

BACKGROUND

Tubulo-interstitial fibrosis (TIF) is the common pathway in the chronic kidney disease (CKD). Epithelial-to-mesenchymal transition (EMT) is a major contributor to the TIF by the increased myofibroblasts. Renin-angiotensin system (RAS) is critical mediator on EMT in progressive CKD. Angiotensin II (ANG) mediates EMT and causes TIF by stimulating transforming growth factor-β1 (TGF-β1). RAS activation could further activate TGF-β1. Inhibition of the RAS is one of the most powerful therapies for progressive CKD. 25-O-methylalisol F (MAF) is a new tetracyclic triterpenoid compound isolated from the Alismatis rhizoma, which is extensively used for anti-hypertensive, diuretic and anti-hyperlipidemic effects.

METHODS

Inhibitory effect of MAF on EMT is investigated in both TGF-β1- and ANG-induced tubular epithelial cells (NRK-52E) and fibroblasts (NRK-49F). Western blot analysis, qRT-PCR, siRNA, immunofluorescence staining and co-immunoprecipitation techniques were used to evaluate the inhibition of MAF on EMT and further revealed the intervention effects on RAS, TGF-β/Smad and Wnt/β-catenin pathways.

RESULTS

MAF treatment significantly inhibited TGF-β1 and ANG-induced expressions of collagen I, fibronectin, α-SMA, vimentin and E-cadherin at both mRNA and protein levels in the NRK-52E and NRK-49F cells. The action mechanism revealed that MAF significantly ameliorated upregulation of angiotensinogen, renin, ACE and AT1R expressions. Further, MAF attenuated upregulation of Smad3 phosphorylation and downregulation of Smad7, but did not affect the phosphorylation of Smad2, PI3K, ERK1/2 and p38 expressions and Smad4 expression in NRK-52E cells. Co-immunoprecipitation analysis indicated that MAF selectively blocked the combination of Smad3 with TGFβRI and Smad3 with SARA without interfering with the Smad2, TGFβRI and SARA interaction. Additionally, MAF suppressed the expressions of Wnt1 and β-catenin as well as its downstream target Snail1, Twist, MMP-7, PAI-1 and FSP1 expressions in NRK-52E cells.

CONCLUSIONS

MAF simultaneously targeted multiple RAS components and it was a novel RAS inhibitor. MAF inhibited EMT by Smad3-specific signaling in the TGF-β/Smad-dependent pathway and Wnt/β-catenin pathway. MAF has an important effect on crosstalk between the TGF-β/Smad and Wnt/β-catenin pathway in EMT process by activation of RAS.

Authors+Show Affiliations

Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Life Science, Northwest University, No. 229 Taibai North Road, Xi'an, Shaanxi 710069, China.Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Life Science, Northwest University, No. 229 Taibai North Road, Xi'an, Shaanxi 710069, China.Xi'an Modern Chemistry Institute, Xi'an, Shaanxi 710065, China.Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Life Science, Northwest University, No. 229 Taibai North Road, Xi'an, Shaanxi 710069, China.Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Life Science, Northwest University, No. 229 Taibai North Road, Xi'an, Shaanxi 710069, China.Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Life Science, Northwest University, No. 229 Taibai North Road, Xi'an, Shaanxi 710069, China. Electronic address: zyy@nwu.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29655688

Citation

Chen, Hua, et al. "Novel RAS Inhibitor 25-O-methylalisol F Attenuates Epithelial-to-mesenchymal Transition and Tubulo-interstitial Fibrosis By Selectively Inhibiting TGF-β-mediated Smad3 Phosphorylation." Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, vol. 42, 2018, pp. 207-218.
Chen H, Yang T, Wang MC, et al. Novel RAS inhibitor 25-O-methylalisol F attenuates epithelial-to-mesenchymal transition and tubulo-interstitial fibrosis by selectively inhibiting TGF-β-mediated Smad3 phosphorylation. Phytomedicine. 2018;42:207-218.
Chen, H., Yang, T., Wang, M. C., Chen, D. Q., Yang, Y., & Zhao, Y. Y. (2018). Novel RAS inhibitor 25-O-methylalisol F attenuates epithelial-to-mesenchymal transition and tubulo-interstitial fibrosis by selectively inhibiting TGF-β-mediated Smad3 phosphorylation. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, 42, 207-218. https://doi.org/10.1016/j.phymed.2018.03.034
Chen H, et al. Novel RAS Inhibitor 25-O-methylalisol F Attenuates Epithelial-to-mesenchymal Transition and Tubulo-interstitial Fibrosis By Selectively Inhibiting TGF-β-mediated Smad3 Phosphorylation. Phytomedicine. 2018 Mar 15;42:207-218. PubMed PMID: 29655688.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Novel RAS inhibitor 25-O-methylalisol F attenuates epithelial-to-mesenchymal transition and tubulo-interstitial fibrosis by selectively inhibiting TGF-β-mediated Smad3 phosphorylation. AU - Chen,Hua, AU - Yang,Tian, AU - Wang,Min-Chang, AU - Chen,Dan-Qian, AU - Yang,Yang, AU - Zhao,Ying-Yong, Y1 - 2018/03/19/ PY - 2017/11/08/received PY - 2018/01/31/revised PY - 2018/03/17/accepted PY - 2018/4/16/entrez PY - 2018/4/16/pubmed PY - 2018/8/16/medline KW - 25-O-methylalisol F KW - Alismatis rhizoma KW - Epithelial-to-mesenchymal transition KW - Kidney proximal tubular epithelial cells KW - Renal fibrosis KW - Renin-angiotensin system SP - 207 EP - 218 JF - Phytomedicine : international journal of phytotherapy and phytopharmacology JO - Phytomedicine VL - 42 N2 - BACKGROUND: Tubulo-interstitial fibrosis (TIF) is the common pathway in the chronic kidney disease (CKD). Epithelial-to-mesenchymal transition (EMT) is a major contributor to the TIF by the increased myofibroblasts. Renin-angiotensin system (RAS) is critical mediator on EMT in progressive CKD. Angiotensin II (ANG) mediates EMT and causes TIF by stimulating transforming growth factor-β1 (TGF-β1). RAS activation could further activate TGF-β1. Inhibition of the RAS is one of the most powerful therapies for progressive CKD. 25-O-methylalisol F (MAF) is a new tetracyclic triterpenoid compound isolated from the Alismatis rhizoma, which is extensively used for anti-hypertensive, diuretic and anti-hyperlipidemic effects. METHODS: Inhibitory effect of MAF on EMT is investigated in both TGF-β1- and ANG-induced tubular epithelial cells (NRK-52E) and fibroblasts (NRK-49F). Western blot analysis, qRT-PCR, siRNA, immunofluorescence staining and co-immunoprecipitation techniques were used to evaluate the inhibition of MAF on EMT and further revealed the intervention effects on RAS, TGF-β/Smad and Wnt/β-catenin pathways. RESULTS: MAF treatment significantly inhibited TGF-β1 and ANG-induced expressions of collagen I, fibronectin, α-SMA, vimentin and E-cadherin at both mRNA and protein levels in the NRK-52E and NRK-49F cells. The action mechanism revealed that MAF significantly ameliorated upregulation of angiotensinogen, renin, ACE and AT1R expressions. Further, MAF attenuated upregulation of Smad3 phosphorylation and downregulation of Smad7, but did not affect the phosphorylation of Smad2, PI3K, ERK1/2 and p38 expressions and Smad4 expression in NRK-52E cells. Co-immunoprecipitation analysis indicated that MAF selectively blocked the combination of Smad3 with TGFβRI and Smad3 with SARA without interfering with the Smad2, TGFβRI and SARA interaction. Additionally, MAF suppressed the expressions of Wnt1 and β-catenin as well as its downstream target Snail1, Twist, MMP-7, PAI-1 and FSP1 expressions in NRK-52E cells. CONCLUSIONS: MAF simultaneously targeted multiple RAS components and it was a novel RAS inhibitor. MAF inhibited EMT by Smad3-specific signaling in the TGF-β/Smad-dependent pathway and Wnt/β-catenin pathway. MAF has an important effect on crosstalk between the TGF-β/Smad and Wnt/β-catenin pathway in EMT process by activation of RAS. SN - 1618-095X UR - https://www.unboundmedicine.com/medline/citation/29655688/Novel_RAS_inhibitor_25_O_methylalisol_F_attenuates_epithelial_to_mesenchymal_transition_and_tubulo_interstitial_fibrosis_by_selectively_inhibiting_TGF_β_mediated_Smad3_phosphorylation_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0944-7113(18)30069-2 DB - PRIME DP - Unbound Medicine ER -