J Wen1, H Zhu1, X Li1, J Huang1, Y Chen1, Q Yang1. 1. Department of Neurology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Abstract
OBJECTIVE: To investigate the effect of TGF-β1 on Shh signaling pathway during the transformation of meningeal fibroblasts into myofibroblasts. METHODS: Primary meningeal fibroblasts were isolated from neonatal (24 h) SD rats and purified using type Ⅳ collagenase. The isolated cells were treated with 10 ng/mL TGF-β1 alone or in combination with 20 μmol/L SB-431542 (a TGF-β1 receptor inhibitor) for 72 h, and the changes in proliferation and migration abilities of the fibroblasts were assessed with CCK-8 assay and cell scratch test. The expression of fibronectin (Fn) was detected with immunofluorescence assay, and Western blotting was performed to examine the expressions of Fn, α-SMA and Shh protein in the cells; the expression of Shh mRNA was detected with real-time fluorescence quantitative PCR. RESULTS: TGF-β1 treatment obviously enhanced the proliferation and migration of primary meningeal fibroblasts (P < 0.05), and promoted the transformation of meningeal fibroblasts into myofibroblasts and the secretion of Fn (P < 0.05). TGF-β1 treatment also upregulated the expression of Shh at both protein and mRNA levels (P < 0.05). Treatment with SB-431542 partially blocked the effect of TGF-β1 on the transformation of meningeal fibroblasts (P < 0.05). CONCLUSION: TGF-β1 can induce the transformation of meningeal fibroblasts into myofibroblasts by up-regulating Shh expression in Sonic Hedgehog signaling pathway.
OBJECTIVE: To investigate the effect of TGF-β1 on Shh signaling pathway during the transformation of meningeal fibroblasts into myofibroblasts. METHODS: Primary meningeal fibroblasts were isolated from neonatal (24 h) SD rats and purified using type Ⅳ collagenase. The isolated cells were treated with 10 ng/mL TGF-β1 alone or in combination with 20 μmol/L SB-431542 (a TGF-β1 receptor inhibitor) for 72 h, and the changes in proliferation and migration abilities of the fibroblasts were assessed with CCK-8 assay and cell scratch test. The expression of fibronectin (Fn) was detected with immunofluorescence assay, and Western blotting was performed to examine the expressions of Fn, α-SMA and Shh protein in the cells; the expression of Shh mRNA was detected with real-time fluorescence quantitative PCR. RESULTS: TGF-β1 treatment obviously enhanced the proliferation and migration of primary meningeal fibroblasts (P < 0.05), and promoted the transformation of meningeal fibroblasts into myofibroblasts and the secretion of Fn (P < 0.05). TGF-β1 treatment also upregulated the expression of Shh at both protein and mRNA levels (P < 0.05). Treatment with SB-431542 partially blocked the effect of TGF-β1 on the transformation of meningeal fibroblasts (P < 0.05). CONCLUSION: TGF-β1 can induce the transformation of meningeal fibroblasts into myofibroblasts by up-regulating Shh expression in Sonic Hedgehog signaling pathway.
Authors: Maria J Guillen-Sacoto; Ariel F Martinez; Yu Abe; Paul Kruszka; Karin Weiss; Joshua L Everson; Ramon Bataller; David E Kleiner; Jerrold M Ward; Kathleen K Sulik; Robert J Lipinski; Benjamin D Solomon; Maximilian Muenke Journal: J Hepatol Date: 2017-06-21 Impact factor: 25.083
Authors: Thomas D Arnold; Richard Daneman; Cayce E Dorrier; Dvir Aran; Ezekiel A Haenelt; Ryan N Sheehy; Kimberly K Hoi; Lucija Pintarić; Yanan Chen; Carlos O Lizama; Kelly M Cautivo; Geoffrey A Weiner; Brian Popko; Stephen P J Fancy Journal: Nat Neurosci Date: 2021-02-01 Impact factor: 24.884