05), while other inhibitors (ERK inhibitor, JNK inhibitor and PKA

05), while other inhibitors (ERK inhibitor, JNK inhibitor and PKA inhibitor) had no effect on the induction of smad7 by exogenous TGF-β3 stimulation (P > 0.05). 6) In basal condition, exogenous TGF-β1 also increased smad7 mRNA expression in HSC (1.5-fold higher than control, P < 0.05), but this induction is lower AG-14699 than it by exogenous TGF-β3. Additionally, the inhibition and over-expression of CREB-1 had no effect on exogenous TGF-β1-induced smad7 expression in HSC (P > 0.05). Conclusion: 1) TGF-β3 increases smad7 expression in HSC. 2) smad3 is an important transcriptional regulator for smad7. 3) CREB-1 is critical for TGF-β3-induced samd7 in HSC. 4) TGF-β3 activates CREB-1

by p38 in HSC. Taken together, TGF-β3 might activate both smad3 and CREB-1, and CREB-1 is an important co-transcriptional factor which enhances the binding of smad3 with DNA, caused a continuous

induction of smad7 in HSC, and CREB-1 might contribute to resist liver fibrosis. Key Word(s): 1. Liver fibrosis; 2. CREB-1; 3. TGF-β3; 4. smad7; Presenting Author: YANHUA SHEN Additional Authors: HAIXING JIANG Corresponding Author: HAIXING JIANG Affiliations: 1st Affiliated hospital of Guangxi medical university Objective: To investigate the effect of activated hepatocyte growth factor (HGF) on hepatic stellate cells (HSCs) apoptosis and the regulation of Rho pathway. Methods: HSCs were divided into the following groups: check details ① the blank control group: HSCs were cultured alone; ② the control group: a. HSCs were cultured with exogenous HGF (50 ng/ml), b. HSCs were cultured with exogenous HGFA (70 ng/ml); ③ the experimental group: HSCs were co-cultured with exogenous HGF and HGFA; ④ HGF inhibitor groups: HSCs were incubated with c-met (500 ng/ml) blockers for 6 hours, and then with exogenous HGF and HGFA; ⑤ Rho pathway inhibitor groups: HSCs were cultured with Y-27632

(10 ng/ml), and then with exogenous HGF and HGFA. The activation of HSC was determined by analysis of alpha smooth muscle actin (α-SMA) expression. The best intervention concentration of Y – 27632 was detected by MTT assay; MCE公司 HSCs apoptosis was tested by Flow Cytometry; the expression of HGF alpha chain was determined by Immunofluorescence; RohA mRNA levels were evaluated by PCR. Protein expressions were evaluated by immunohistochemical staining and Western blot analysis. Results: ① Y-27632 at 10 μ mol/L caused obviously HSCs inhibition (P < 0.01) compared with other concentration groups. ② The expression of the HGF-α chain showed time-dependent increased manner (P < 0.01). However, there was no statistic difference (P > 0.05) in blank control group and control group. ③ The apoptosis rate increased over time (24 h, 48 h, 72 h) (P < 0.01). The experimental group caused the highest levels (P < 0.01). ④ The expression of RhoA mRNA in experimental group decreased over time (P < 0.01) and caused the lowest levels compared with othergroups (P < 0.01).

Leave a Reply

Your email address will not be published. Required fields are marked *


You may use these HTML tags and attributes: <a href="" title=""> <abbr title=""> <acronym title=""> <b> <blockquote cite=""> <cite> <code> <del datetime=""> <em> <i> <q cite=""> <strike> <strong>