RESUMO
BACKGROUND: Pancreatic stellate cells (PSCs) play a critical role in the development of pancreatic fibrosis. In this study we used a novel method to isolate and culture rat PSCs and then investigated the inhibitory effects of adipose-derived stem cells (ADSCs) on activation and proliferation of PSCs. METHODS: Pancreatic tissue was obtained from Sprague-Dawley rats for PSCs isolation. Transwell cell cultures were adopted for co-culture of ADSCs and PSCs. PSCs proliferation and apoptosis were determined using CCK-8 and flow cytometry, respectively. alpha-SMA expressions were analyzed using Western blotting. The levels of cytokines [nerve growth factor (NGF), interleukin-10 (IL-10) and transforming growth factor-beta1 (TGF-beta1)] in conditioned medium were detected by ELISA. Gene expression (MMP-2, MMP-9 and TIMP-1) was analyzed using qRT-PCR. RESULTS: This method produced 17.6+/-6.5X10(3) cells per gram of the body weight with a purity of 90%-95% and a viability of 92%-97%. Co-culture of PSCs with ADSCs significantly inhibited PSCs proliferation and induced PSCs apoptosis. Moreover, alpha-SMA expression was significantly reduced in PSCs+ADSCs compared with that in PSC-only cultures, while expression of fibrinolytic proteins (e.g., MMP-2 and MMP-9) was up-regulated and anti-fibrinolytic protein (TIMP-1) was down-regulated. In addition, NGF expression was up-regulated, but IL-10 and TGF-beta1 expressions were down-regulated in the co-culture conditioned medium compared with those in the PSC-only culture medium. CONCLUSIONS: This study provided an easy and reliable technique to isolate PSCs. The data demonstrated the inhibitory effects of ADSCs on the activation and proliferation of PSCs in vitro.
Assuntos
Tecido Adiposo/citologia , Apoptose , Proliferação de Células , Células Estreladas do Pâncreas/fisiologia , Células-Tronco , Actinas/metabolismo , Animais , Técnicas de Cocultura , Meios de Cultivo Condicionados , Regulação para Baixo/genética , Expressão Gênica , Interleucina-10/metabolismo , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Fator de Crescimento Neural/metabolismo , Ratos , Ratos Sprague-Dawley , Inibidor Tecidual de Metaloproteinase-1/genética , Fator de Crescimento Transformador beta1/metabolismo , Regulação para Cima/genéticaRESUMO
BACKGROUND: Extensive controversies exist over the use of preoperative biliary drainage preceding radical resection of hilar cholangiocarcinoma. This study assessed the effectiveness and safety of percutaneous transhepatic cholangiodrainage (PTCD) with bile re-infusion in the preoperative optimization of hilar cholangiocarcinoma patients. METHODS: Eligible hilar cholangiocarcinoma patients received preoperative PTCD with bile re-infusion (treatment group, n = 56) through a nasoduodenal tube for 2 weeks, and the control group (n = 60) received conservative treatment alone. Operable patients were assigned to undergo either a radical or palliative resection. The outcome measures included the overall resection rate, R0 resection rate, surgical morbidity rate and 1-year and 5-year overall survival rates. RESULTS: The treatment group exhibited a significant decrease in serum bilirubin levels after PTCD with bile re-infusion. The overall resection rate was significantly higher in the treatment group than in the control group (85.5% vs. 65.0%, P < 0.05), and the palliative resection rate was also significantly higher in the treatment group (53.5% vs. 35.0%, P < 0.05). However, the R0 resection rate was comparable between the 2 groups (32.1% vs. 30.0%, P > 0.05). The morbidity rate was significantly lower in the treatment group than in the control group (29.1% vs. 51.3%, P < 0.05). One-year and 5-year survival rates were similar between the 2 groups (69.6% vs. 66.7%, P > 0.05; 5.3% vs. 3.6%, P > 0.05). CONCLUSIONS: Preoperative PTCD with bile re-infusion improves the resection rate and shows a good safety profile in patients with hilar cholangiocarcinoma.