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5‐ HT 2 and 5‐ HT 2B antagonists attenuate pro‐fibrotic phenotype in human adult dermal fibroblasts by blocking TGF ‐β1 induced non‐canonical signaling pathways including STAT 3 : implications for fibrotic diseases like scleroderma
Author(s) -
Chaturvedi Saurabh,
Misra Durga Prasanna,
Prasad Narayan,
Rastogi Kailash,
Singh Harshit,
Rai Mohit Kumar,
Agarwal Vikas
Publication year - 2018
Publication title -
international journal of rheumatic diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 41
eISSN - 1756-185X
pISSN - 1756-1841
DOI - 10.1111/1756-185x.13386
Subject(s) - ctgf , extracellular matrix , fibrosis , medicine , fibroblast , transforming growth factor , mapk/erk pathway , ed50 , pharmacology , microbiology and biotechnology , endocrinology , growth factor , andrology , chemistry , receptor , phosphorylation , biology , biochemistry , in vitro
Background Release of 5‐hydroxytryptamine (5‐ HT ; serotonin) from activated platelets following microvascular injury leads to tissue fibrosis. 5‐ HT strongly induces extracellular matrix synthesis in dermal fibroblasts in a transforming growth factor beta 1 ( TGF ‐β1)‐dependent manner. Aim To evaluate anti‐fibrotic properties of inhibitors of 5‐ HT 2 and 5‐ HT 2B (terguride, SB 204741) respectively in human adult dermal fibroblasts ( HADF ) derived from a patient with scleroderma. Methods Anti‐fibrotic efficacy of 5‐ HT 2 and 5‐ HT 2B inhibitors was evaluated as per two strategies: HADF were incubated with 5‐ HT (1 μM)/ TGF ‐β1 (10 ng/ mL ) for 1 hour followed by 5‐ HT (1 μM)/ TGF ‐β1 (10 ng/ mL ) and terguride or SB 204741 (1 μM, each) for 24 hours (post‐treatment strategy) and HADF were treated with terguride or SB 204741 (1 μM, each) for 1 hour followed by 5‐ HT (1 μM)/ TGF ‐β1 (10 ng/ mL ) for 24 hours (pre‐treatment strategy). Real time quantitative polymerase chain reaction for expression of pro‐fibrotic ( TGF Β1 , COL 1A1 , COL 1A2 , ACTA 2 , CTGF and FN 1 ) and anti‐fibrotic genes ( MMP 2 / TIMP 1 ) was performed. Expression of type I collagen, alpha smooth muscle actin (α‐ SMA ), phosphorylation of Smad3, ERK 1/2 and STAT 3 was examined by immunoblotting. Results Stimulation of HADF cells with 5‐ HT / TGF ‐β1 led to the increased expression of pro‐fibrotic genes which was significantly reduced by both terguride and SB 204741. Expression of anti‐fibrotic genes was not affected upon incubation with the inhibitors. In 5‐ HT ‐stimulated HADF , treatment with terguride and SB 204741 decreased type I collagen and α‐ SMA . In 5‐ HT / TGF ‐β1 stimulated HADF , terguride and SB 204741 treatment reduced ERK 1/2 and STAT 3 phosphorylation but did not influence Smad3 phosphorylation. Conclusion Terguride and SB 204741 reduce pro‐fibrotic potential of HADF cells and suppress TGF ‐β1‐mediated non‐canonical pathways, ERK 1/2 and STAT 3 which have been implicated in the regulation of pro‐fibrotic genes and in the development of fibrosis. Taken together, our data suggest that 5‐HT inhibitors might reduce fibrosis via suppression of TGF‐beta1‐mediated non‐canonical signaling pathways. These observations have important therapeutic implications for fibrotic disorders like scleroderma.