Differentiation of hepatocytoid cell induced from whole-bone-marrow method isolated rat myeloid mesenchymal stem cells.

AIM: To explore the expansion and differentiation of hepatocytoid cell induced from myeloid mesenchymal stem cell (MSC) in vitro, in order to find suitable resource of hepatocytes for bioartificial liver or liver transplantation. METHODS: The rat myeloid MSC was isolated and divided into three groups which were cultured by Friedensteion method, and then were induced by culture fluid, culture fluid plus cholestatic serum and culture fluid plus hepatocyte growth factor (HGF), respectively. Hepatocytoid cell as well as expression of CK18 and AFP was observed by immunohistochemistry. RESULTS: After the induction for 21 d, hepatocytoid cell was observed, and its expression of CK18 and AFP was detected by immunohistochemistry in MSC cultured with cholestatic serum. Furthermore, on the 35th d, albumin mRNA was expressed in the cell, suggesting the inducing effect was similar to that by HGF. CONCLUSION: Rat myeloid MSC can differentiate into hepatocyte lineage under appropriate condition. This method is easy to operate.

[Study on the effect of heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice].

OBJECTIVE: To investigate the effect of Chinese traditional medicine heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice. METHODS: Liver fibrosis in mice was established by Schistosoma japonicum infection in 6 weeks. Suspension of heluoshugan prepared with normal saline was given orally to the mice, 2 capsules for 20 mice daily for 8 weeks. The level of vascular endothelial growth factor (VEGF), focal adhesion kinase (FAK) and type I, III collagen in liver tissue were detected by immuno-histochemistry. RESULTS: The results showed that heluoshugan improved the pathological change of the liver tissue, decreased the level of type I, III collagen, especially type III collagen (P < 0.01). The level of VEGF and FAK expression was inhibited after the administration of heluoshugan, though the level usually increased in liver fibrosis due to the infection. CONCLUSIONS: The result suggests that heluoshugan capsule might have therapeutic effect on liver fibrosis induced by the infection of Schistosoma japonicum in mice, by inhibiting the activation of hepatic stellate cells and the pathological change of liver blood vessel.

[In vitro study on Hep G2 cell infected by hepatitis C virus].

OBJECTIVE: To establish hepatitis C virus (HCV) infected cell model which is similar to the infection in vivo and can support HCV to replicate for a long time. METHODS: After infected with HCV-positive serum, Hep G2 cells were cultured for 60 days. Nested RT-PCR was used to detect plus and minus HCV RNA in cultured cells and supernatants. RESULTS: Plus HCV RNA was detected intermittently in Hep G2 cells during 2-30 days, minus HCV RNA was detected during 3-30 days after infection, the detection rate was similar to plus HCV RNA. Plus and minus HCV RNA can be still intermittently detected during 31-60 days after infection. However, the detection rate gradually declined. Plus HCV RNA was also found intermittently positive in the supernatant, and the detection rate was consistent to that in cells. Minus HCV RNA was not detected in the supernatant. CONCLUSION: Hep G2 cells were susceptible to HCV, and could support HCV to replicate for a relatively long time. Hep G2 is an ideal HCV infection cell model.

[Expression of hepatic Bcl-2 and Bax proteins in schistosome-infected mice and the role of pentoxifylline].

OBJECTIVE: To study the expression of hepatic Bcl-2 and Bax proteins in mice infected with Schistosoma japonicum and the role of pentoxifylline (PTX) in the expression. METHODS: Forty mice were randomly divided into 4 groups: one normal control group,mice in the other three groups were all infected each with 25 cercariae, the infected control group was fed for 10 weeks after infection, and 2 weeks after infection, the high dose PTX group was given PTX 360 mg/(kg x d) for 8 weeks and the low dose PTX group was given PTX 180 mg/(kg x d)also for 8 weeks. At the end of 10 weeks all the mice were killed. Bcl-2 and Bax proteins expression was detected by immunohistochemisty. RESULTS: Compared with the normal control group, the expression of Bcl-2 and Bax was significantly higher in the infected control group (P < 0.05). Bcl-2 was significantly higher in high (dose PTX group than in the infected control group and in low dose PTX group (P < 0.05). However there was no significant difference in the expression of Bax among the groups (P > 0.05). CONCLUSION: PTX treatment can significantly increase the expression of Bcl-2 in liver tissue of schistosome-infected mice in a dose-dependent manner, and may play a role against liver inflammation and schistosomiasis-related liver fibrosis.

[Expression of IL-2 and TNF-alpha in the liver and the effect of injection of these cytokines on liver fibrosis in mice infected with Schistosoma japonicum].

OBJECTIVE: To detect the expression of IL-2 and TNF-alpha in the liver at different period postinfection of Schistosoma japonicum and their effect on liver fibrosis after supplementary injection of these cytokines. METHODS: Mice were infected with schistosome cercariae and divided into 3 groups. Two groups were injected (i.p.) every other day with IL-2 and TNF-alpha respectively for consecutive 4 wk. The third group and an uninfected group of normal mice were regarded as control. The ABC immunohistochemistry and pathologic image multimedia quantification system were applied to detect activity of IL-2 and TNF-alpha. RESULTS: The level of IL-2 and TNF-alpha in the liver in infected but untreated group slowly decreased (from 8, 11, 14 to 18 wk). The supplementary injection of the cytokines at 6 wk postinfection in the two groups increased the cytokines significantly, the level of IL-2 or TNF-alpha was higher at 1-8 wk after the last injection than that of both infected and uninfected control groups (P < 0.01). The granulomatous inflammation and fibrosis in the livers of the two groups were slighter than that of the control. CONCLUSION: At the 6th wk postinfection with egg deposition, exogenous supplementation with TNF-alpha or IL-2 induces enhanced expression of the two kinds of cytokines, corresponding to a diminished degree of the liver granulomatous inflammation and fibrosis.

Effects of pentoxifylline on the hepatic content of TGF-beta1 and collagen in Schistosomiasis japonica mice with liver fibrosis.

AIM: To study the effects of pentoxifylline (PTX) on the content of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis and its mechanism of anti-fibrosis. METHODS: Forty mice with schistosomiasis were divided into four groups: one group as control without any treatment, other three were treated with Praziquantel 500 mg/(kg x d)for 2 d, high dose PTX 360 mg/(kg x d) for 8 wk, and low dose PTX 180 mg/(kg x d) for 8 wk respectively. Immunohistochemical technique and multimedia color pathographic analysis system were applied to observe the content change of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis before and after PTX treatment. RESULTS: Effects of PTX on the content change of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis were related to the dosage of PTX, high dose PTX treated group could significantly reduce the content of TGF-beta1 (0.709+/-0.111), type I (0.644+/-0.108) and type III (0.654+/-0.152) collagen compared with those of control group (0.883+/-0.140, 0.771+/-0.156, 0.822+/-0.129) with statistical significance (P<0.05). Low dose PTX could also reduce the hepatic content of TGF-beta1 (0.752+/-0.152), type I (0.733+/-0.117) and type III (0.788+/-0.147) collagen, but without statistical significance (P>0.05). Both high dose and low dose PTX groups have significant differences on the content of TGF-beta1, type I and type III collagen (P<0.05, P<0.05, P<0.01, respectively). CONCLUSION: High dose of PTX treatment could reduce the content of hepatic TGF-beta1, type I and type III collagen significantly in schistosomiasis japonica mice with liver fibrosis, and thus plays its role of antifibrosis.