However whether these cells also interact with hepatic stellate cells (HSCs) and contribute to liver fibrosis remains unknown. In the current study, we evaluated the effect of EPC-secreted angiogenic factors on the functions of HSCs under in vitro conditions and also the effect of EPC transplantation on liver fibrosis in bile duct ligated (BDL) cirrhotic rat models in vivo. Methodology: For the in vitro studies, HSCs were isolated from normal rats after liver perfusion, cultured and characterized by alpha-SMA staining. Circulating human EPCs were isolated from blood and characterized by CD34 and

vegfr2 staining. HSCs were then co-cultured with conditioned this website media (CM) obtained from EPCs. The

proliferation of HSCs was analyzed by MTT assay and the level of an important angiogenic factor secreted by HSCs, vascular endothelial growth factor (VEGF) was evaluated by ELISA in presence of EPC-CM. For in vivo studies, rat models (n= 1 0) were Selleck R788 prepared by identifying and ligating the bile duct. EPCs isolated from human blood, were cultured ex vivo and transplanted in treated group of BDL rats (n=5) via the tail vein, three weeks after bile duct ligation. The untreated group of rats (n= 5) received only saline. Rats were sacrificed one week after the transplantation of cells. Fibrosis was evaluated by histopathology of the liver tissues from untreated and EPC-treated rats. The expression of an important fibrogenic marker, alpha-SMA was analyzed in EPC-treated and untreated animals by western blotting. Results: HSCs co-cultured with EPC-CM showed significantly increased proliferation in comparison to that observed in HSCs alone (P< 0.05).

However, VEGF levels in HSCs didn’t show a significant change in presence of EPC-CM. The in vivo histopathology studies revealed an increase of fibrosis from stage 2 to stage 3 (bridging fibrosis) in EPC-treated rats as compared to the untreated rats. Also, the expression of the fibrosis marker, alpha-SMA present on activated HSCs was about 1.2 fold higher in EPC-treated rats as compared to the untreated rats (p<0.05). Conclusion: The study suggests that EPCs may contribute to liver fibrosis by enhancing the proliferation of HSCs. Further Afatinib purchase studies are underway to evaluate the association of EPC-mediated angiogenesis with liver fibrosis. Disclosures: The following people have nothing to disclose: Skand Gupt, Mohsin Hasan, Neha Sharma, Vaibhav Dixit, Deepti Vyas, Dinesh M. Tripathi, Savneet Kaur, Nirupma Trehanpati, Shiv K. Sarin Background and aims: 1,25(OH)2D3, the active form of vitamin D has anti-proliferative and anti-fibrotic effect on hepatic stellate cells. Our aim was to investigate the potential of 1,25(OH)2D3 to inhibit the development of liver fibrosis and to ameliorate established fibrosis in vivo.