Using transwell insert establish co-culture system in the

plastic plate, the cells were observed dynamically under inverted phase contrast microscope after 24, 48 and 72 h. Expression of alpha smooth muscle actin (α-SMA) in HSCs were evaluated by immunohistochemistry. The best intervention concentrations of Y-27632 and PHA665752 were determined by MTT assay. The apoptosis rate of HSCs were measured by Annexin-V-FITC/propidium iodide (PI). RohA mRNA LBH589 clinical trial and protein levels were measured by quantitative real time polymerase chain reaction (Q-PCR) and Western blot, respectively. The concentration of HGF and HGFA were quantified by enzyme-linked immunosorbent assay (ELISA). Results: ○1Under Inverted phase contrast microscope cells were observe the good condition of BMSCs performance large cell body, refract well, a typical long spindle; GSI-IX nmr good condition of HSCs was membrane growth, typical star

or polygon, intracellular more grain. ○2Cultured for 48 h, brown granules were viewed in the cytoplasm within HSCs and light blue nuclear. The results show α-SMA(+) and More than 94% of activated HSCs positive. ○1The apoptosis rate of HSCs gradually increased at all time points examined, the apoptosis rate of the PHA665752 pretreated group was lowest, but the Y-27632 pretreated group was highest, most significant in 72 h (P < 0.05). ○2The expression of RhoA mRNA and proteins in Y-27632 pretreated group significant decrease over time (24,48,72 h) compared with other groups (P < 0.01) and the expression of RhoA mRNA and proteins increased over time (P < 0.01).○3The concentration of HGF in experimental

groups decrease over time (24 h,48 h,72 h), the PHA665752 pretreated group and the Y-27632 pretreated group were significant higher than the control group (P < 0.05). The concentration of HGFA increase over time (24 h,48 h,72 h), the concentration of HGFA in the PHA665752 pretreated group was higher than any other groups at any time (P < 0.01). ○6 Y-27632 at 30 μmol/L and Dolichyl-phosphate-mannose-protein mannosyltransferase PHA665752 at 3 μg/ml caused obviously HSCs apoptosis (P < 0.05). Conclusion: BMSCs promoted HSCs apoptosis by activating HGF and downregulating RhoA signaling pathway. Key Word(s): 1. HSC; 2. BMSC; 3. HGF; 4. RhoA; Presenting Author: 茜 Corresponding Author: 茜 Affiliations: none Objective: End-stage liver disease (ELD) is the common pathway of the acute or chronic liver disease in process. Hepatic stellate cells (HSCs) play a vital role in the development and progression of various liver disease. HSCs are the main extracellular matrix synthesis cells, which its activiation and transformation play an important role in liver cirrhosis. Currently, the treatment for ELD is limited, and orthotopic liver transplantation (OLT) may be the best choice. However, OLT has its limitation by that extreme short of donor liver, expensive cost of operation and severe rejection of transplantation.

No related posts.