| Due to the stimulation of a variety of cytokines, particularly in chronic hepatitis stage causing by the hepatitis B virus infection, intrahepatic stellate cells are activated, besides, the liver extracellular matrix is synthesized in large quantities, resulting in a large number of fibrous tissue deposition and the formation of liver fibrosis. Hepatitis B virus itself does not cause damage to the liver, what mainly cause liver damage is the host-specific immune responses. Cytotoxic T lymphocyte not only can clear the Hepatitis B virus, but also can kill the infected hepatocyte.We combined mesenchymal stem cells transplantation with RNA interference to treat liver injury. RNA interference (RNAi), by which a small interfering RNA (siRNA) induces the gene silence at a post-transcriptional level, has the potential of treating HBV infection. Mesenchymal stem cells (MSCs) have a wide rang of sources and no immunogen, which are easy to obtain. It has been reported both in vivo and in vitro MSCs have the potential to differentiate into hepatocytes. MSCs can also migrate to the injured liver tissue to repair it. After transplantation, mesenchymal stem cells can secrete some cytokines which can also help to recovery the host liver injury. Therefore, MSCs are the ideal donor of liver transplantation.It has been reported at home and abroad that RNA interference technology can inhibited HBV replication. However, for patients with chronic HBV infection in midanaphase, although HBV replication was inhibited by RNA interference, the degree of liver damage is very serious and liver cells can’t repair themselves. On the one hand, we use targeting shRNA to inhibit HBV replication in vivo. On the other hand, transplantation of MSCs by intrasplenic injection can repair liver injury. Thus, we not only inhibited HBV replication, but also improved liver injury.First, according to literatures we chose different HBV DNA targets. Lx-shRNA157expression plasmids target on the site of HBV DNA genome S open reading frame157i. Lx shRNA1694expression plasmids target on the site of HBV DNA genome X open reading frame1694i. Lx-shRNA157-1694expression plasmids target on double sites. We have synthesized three shRNA expression plasmids, which all have good inhibition effects on HBV replication. And no matter in vivo or in vitro Lx-shRNA157-1694have better inhibition effects on HBV replication than a single target. Secondly, through the special culture mediums, we successfully differentiated HUC-MSCs and mi-MSCs into bone cells and liver cells in vitro. As the same time, we have proved that after MSCs differentiated into liver cells, they have the functions of mature hepatocytes, such as glycogen storage and urea synthesis.It’s difficult to research HBV, because the host range of HBV is narrow. It is difficult to find a suitable animal model. Finally, we have successfully constructed the hepatitis B virus liver fibrosis model in mice. The mice model both have HBV replication and liver fibrosis, in order to simulate the intracorporal environment of chronic HBV infected patients. We combined shRNA and MSCs transplantation technology for the treatment of liver injury, as shRNA can specifically inhibit HBV replication and MSCs can repair the liver tissue injury. Then we inhibited HBV replication and restored normal function of liver cells, so as to treat the chronic HBV infected patients.This paper have verified the functions of shRNA and MSCs through the above experiments. The combination of these two technologies can be more effective in treating liver injury caused by HBV infection, which can provide a new therapeutic strategy for the treatment of HBV infection. |
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