[Changes of Toll-like Receptor mRNA and Related Cytokines in Patients with Hepatic Alveolar Echinococcosis].

Objective: To investigate the expression of Toll-like receptor 2（TLR2） and TLR4 mRNA in peripheral blood mononuclear cells （PBMC） and in the liver of patients with hepatic alveolar echinococcosis (HAE), and their correlations with related cytokines in plasma. Methods: Twenty-eight HAE patients hospitalized in the First Affiliated Hospital of Xinjiang Medical University during January 2012 and June 2015 and 28 healthy volunteers as a control were enrolled in this study. Plasma levels of interferon-γ （IFN-γ）, interleukin-5 （IL-5）, IL-23, and IL-10 were measured by ELISA. qRT-PCR was performed to detect TLR2 and TLR4 mRNA levels in PBMCs and hepatic tissues. The percentage of peripheral blood eosinophil （Eo%） was determined by a hematology analyzer. The correlations of TLR2 and TLR4 mRNA levels in PBMCs with levels of related cytokines and Eo% were analyzed with the Spearman Correlation method. Results: ELISA results showed that the plasma levels of IFN-γ, IL-5, IL-23, and IL-10 in the HAE group were （301.100±47.290）, （43.420±11.380）, （86.580±31.990） and （8.766±7.568） pg/ml respectively, which were higher than those in the control［（301.100±67.790）, （40.970±6.310）, （46.770±15.490） and （6.272±10.360） pg/mlï¼½ with a statistical significance for IL-23 （P<0.01）. Results of qRT-PCR showed that the expression level of TLR2 in the HAE group （0.100±0.084） was significantly higher than that in the control （0.055±0.040） （P<0.05）, while the expression level of TLR4 in the HAE group （0.004±0.003） was comparable to that in the control（0.003±0.002）（P>0.05）. The expression of TLR2 and TLR4 mRNA in HAE lesions in the HAE group（29.680±25.650 and 21.340±16.640, respectively） were both significantly higher than that in para-lesion regions（2.308±4.140 and 5.541±9.233） and that in tissues of the control （1.112±1.431 and 1.100±1.734）（P<0.01）. There was also a significant difference in Eo% between the HAE（0.448±0.240） and the control（0.110±0.100） groups. Spearman correlation coefficients revealed a positive correlation of TLR2 mRNA in PBMCs with plasma IL-23 level and peripheral blood Eo% in HAE subjects（r=0.368, r=0.382, respectively）. Conclusion: There are increases in TLR2 and TLR4 mNRA expression in PBMCs and in HAE lesions in HAE patients. The TLR2 mNRA expression in PBMCs positively correlates with plasma IL-23 level and peripheral Eo%.

TGF-ß1 signaling activates hepatic stellate cells through Notch pathway.

Hepatic stellate cells (HSCs), as the most important stromal cells in the liver microenvironment, play crucial roles in hepatic fibrosis, hepatocellular carcinoma, liver regeneration and fetal liver development after transdifferentiating into myofibroblasts (MFs). Transforming growth factor ß1 (TGF-ß1), as an important polyergic cytokine, is involved in HSCs activation process. However, the specific mechanisms of HSCs transdifferentiation process are not clearly demonstrated. Here we added exogenous recombinant TGF-ß1 protein and transforming growth factor ß receptor 1 (TGF-ßR1) inhibitor SB431542 into mouse HSCs to detect the detailed impact of TGF-ß1 signaling on HSCs activation. TGF-ß1 signaling significantly increased phosphorylated (P)-Smad2/3 level and promoted Smad2/3 translocation from the cytoplasm to the nucleus, which also caused transdifferentiation of HSCs into MFs. Importantly, TGF-ß1 signaling also resulted in high expression of Notch pathway markers Notch1, Jagged1, Hes1 in HSCs. In contrast, expression of those above markers in mouse HSCs were obviously decreased after hampering TGF-ß1 signaling via TGF-ßR1 inhibitor SB431542. To further examine the effect of Notch pathway on HSCs activation process, TGF-ß1-stimulated HSCs and control HSCs were treated with or without LY450139, a specific inhibitor of Notch pathway. LY450139 evidently decreased the expression of Notch1 and MFs marker α-smooth muscle actin (α-SMA) expression in HSCs. These above results may provide a novel insight that TGF-ß1 signaling controls HSCs activation process through regulating the expression of Notch pathway markers.