Abstract:We observed the impact of the expression of TGF-β1 and collagen Ⅰ, Ⅲ in hepatic stellate cells in ICOS transgenic mice infected with Schistosoma japonicum, and explored the new effective way for controlling the development of hepatic fibrosis. ICOS transgenic (ICOS-Tg) mice and wild-type mice as experimental schistosomiasis model for Schistosoma japonicum were established. The liver of ICOS-Tg mice was perfused in situ successively under agitation. HSCs were then separated by density gradient centrifugation through 16% and 12% OptiPrep on day before infection 0 week, and at 4, 6 and 9 weeks post-infection. Cells viability was tested by erythrosin exclusion. The purity of HSCs was assessed by fluorescence of retinoid-containing vacuoles under ultraviolet excitation and immunocytochemisy of GFAP, which was a specific marker of HSCs not expressed in other liver cell types. Total RNA of primary HSCs cultured for 7 days was extracted by using TRIzol reagent. Real-time quantitative PCR was performed using SYBR Premix Ex Taq to assess the expression levels in primary HSCs Collagen-Ⅰ, Collagen-Ⅲ, α-SMA, TGF-β1, and other genes in the development of schistosomiasis. Purity of HSCs isolates was higher than 90%, and cell viability exceeded 95%. Cells were in good condition during the primary culture. With the develop of schistosomiasis, the degree of activation of HSCs was increasing, and the expression of Collagen-Ⅰ, Collagen-Ⅲ, α-SMA, and TGF-β1 in HSCs were enhancing. The level of HSCs activation in ICOS-Tg mice was significantly higher than that in wild-type mice (P<0.01). Also, the expression on Collagen-Ⅰ, Collagen-Ⅲ, α-SMA, and TGF-β1 in HSCs of ICOS-Tg mice were higher than that in wild-type mice (P<0.01-0.05). The co-stimulatory molecules ICOSL/ICOS signaling pathway has an important impact on the process of HSCs activation and fibrosis formation in mice infected with Schistosoma japonicum.
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