ABSTRACT
Studies have shown that human umbilical cord mesenchymal stem cells (hUCMSCs) could ameliorate liver fibrosis (LF) through inhibiting the activation of hepatic stellate cells (HSCs). However, the specific mechanisms have not been studied clearly. The purpose of this study was to explore the possible mechanism of hepatocyte growth factor (HGF)-transfected hUCMSCs in inhibiting the proliferation and activation of HSCs-T6. The upper and lower double-cell coculture system was established among HGF-hUCMSCs, LV5-NC-hUCMSCs, hUCMSCs, and HSCs-T6 in experimental groups; HSCs-T6 were cultured alone as control group. After coculturing for 1, 2, and 3 days, results showed that HGF-transfected hUCMSCs could decrease cell viability of HSCs-T6 and promote apoptosis; inhibit their activation and reduce the expression of Collagen I, Collagen III, TGF-ß1, Smad2 and Smad3, which may be related to inhibiting the activation of TGF-ß1/Smads signaling pathway. These findings suggested that HGF-transfected hUCMSCs may be used as an alternative and novel therapeutic approach for the treatment of LF.
Subject(s)
Hepatic Stellate Cells , Hepatocyte Growth Factor , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Smad Proteins , Transforming Growth Factor beta1 , Cell Proliferation , Hepatic Stellate Cells/metabolism , Hepatocyte Growth Factor/administration & dosage , Hepatocyte Growth Factor/metabolism , Humans , Liver Cirrhosis/metabolism , Liver Cirrhosis/therapy , Signal Transduction , Smad Proteins/metabolism , Transforming Growth Factor beta1/metabolism , Umbilical CordABSTRACT
The research on human umbilical cord-derived mesenchymal stem cells (hUCMSCs) suggests promising therapeutic strategy for ameliorating liver fibrosis and it can be an effective alternative method of orthotopic liver transplantation. Hepatocyte growth factor (HGF) is the most basic cytokine involved in the inhibition of liver fibrosis and promotion of hepatocyte proliferation and regeneration. The objective of this study was to determine the possible mechanism about how the microencapsulated hUCMSCs made by alginate-poly-lysine-alginate (A-P-A) transfected with HGF could ameliorate liver fibrosis through the TGF-ß1/Smad signaling pathway. The microencapsulated cells were divided into four groups: hUCMSC (microcapsules of hUCMSCs), HGF (microcapsules of HGF+hUCMSCs), LV5-NC (microcapsules of LV5-NC, an rLV-EF1a-EGFP+Puro control lentiviral vector+hUCMSCs), and empty microcapsule (microcapsules without any hUCMSCs), and then transplanted by intraperitoneal injection into carbon tetrachloride (CCl4)-induced liver fibrosis rats, respectively. The results showed that the fibrosis in the hUCMSC, LV5-NC, and HGF groups was significantly alleviated. Moreover, the messenger RNA (mRNA) and protein levels of collagen I, collagen III, α-SMA, TGF-ß1, Smad2, and Smad3 were significantly decreased compared with the empty microcapsule group and these indices in HGF group were more decreased compared with hUCMSC and LV5-NC groups. This study indicated that microencapsulated hUCMSCs transfected with HGF could effectively improve CCl4-induced rat liver fibrosis and the possible mechanism was closely related to the inhibition of TGF-ß1/Smad signaling pathway.
