Jiaquan Huang

IL-17 contributes to autoimmune hepatitis

SummaryThe role of interleukin-17 (IL-17) in autoimmune hepatitis (AIH) was investigated. A mouse model of experimental autoimmune hepatitis was established, and the syngeneic S-100 antigen emulsified in complete Freud’s adjuvant was injected intraperitoneally into adult male C57BL/6 mice. The IL-17 expression in serum and the livers of the mice models was detected by using ELISA and immunohistochemistry, respectively. IL-17 neutralizing antibody was used to study the biological effect of IL-17 in the experimental AIH. IL-17 neutralizing antibody in vivo administration alleviated the hepatic inflammation and ALT level in the AIH model. IL-17 in the peripheral blood mononuclear cells (PBMC) of AIH patients was measured by using real-time PCR method. The results showed that IL-17 level was significantly up-regulated in AIH patients and mice models. It was concluded that IL-17 contributed to the development of AIH and might be a potential therapeutic target of AIH.The role of interleukin-17 (IL-17) in autoimmune hepatitis (AIH) was investigated. A mouse model of experimental autoimmune hepatitis was established, and the syngeneic S-100 antigen emulsified in complete Freud’s adjuvant was injected intraperitoneally into adult male C57BL/6 mice. The IL-17 expression in serum and the livers of the mice models was detected by using ELISA and immunohistochemistry, respectively. IL-17 neutralizing antibody was used to study the biological effect of IL-17 in the experimental AIH. IL-17 neutralizing antibody in vivo administration alleviated the hepatic inflammation and ALT level in the AIH model. IL-17 in the peripheral blood mononuclear cells (PBMC) of AIH patients was measured by using real-time PCR method. The results showed that IL-17 level was significantly up-regulated in AIH patients and mice models. It was concluded that IL-17 contributed to the development of AIH and might be a potential therapeutic target of AIH.

Expression of heat shock protein 47, transforming growth factor-beta 1, and connective tissue growth factor in liver tissue of patients with Schistosoma japonicum-induced hepatic fibrosis.

SUMMARY To detect the expression of pro-fibrotic molecules, such as heat shock protein 47 (Hsp47), transforming growth factor-beta 1 (TGF-β1) and connective tissue growth factor (CTGF) in liver specimens, and analyse their correlations with the progression of schistosomal hepatic fibrosis, liver biopsy was performed in 42 chronic schistosomiasis (CS) patients, 16 chronic hepatitis B (CHB) patients and five healthy individuals (HI). Immunohistochemistry (IHC) analyses displayed that the expression of Hsp47, TGF-β1 and CTGF was increased in CS and CHB patients compared with HI. Using real-time PCR, the mRNA levels of Hsp47, TGF-β1 and CTGF were higher in CS patients compared with HI. In CS patients, the mRNA levels of these genes were correlated with the stage of fibrosis, and TGF-β1 mRNA expression was associated with the grade of inflammation. Additional analyses indicated that the mRNA levels of Hsp47 and CTGF were highly correlated with liver stiffness value and spleen thickness diameter, both of which represented the severity of fibrosis. In conclusion, the three molecules are involved in the pathogenesis of hepatic fibrosis infected by Schistosoma japonicum. TGF-β1 participates not only in the inflammatory process, but also in the fibrotic process in which Hsp47 and CTGF probably play a key role.

Involvement of heat shock protein 47 in Schistosoma japonicum-induced hepatic fibrosis in mice

Chronic infection with the blood fluke Schistosoma japonicum is associated with both liver cirrhosis and liver cancer. Previously, heat shock protein 47, a collagen-specific molecular chaperone, was shown to play a critical role in the maturation of procollagen. However, less is known about the role of heat shock protein 47 in S. japonicum-induced hepatic fibrosis. We therefore investigated the expression of heat shock protein 47 in S. japonicum-induced liver fibrosis and attempted to determine whether inhibition of heat shock protein 47 could have beneficial effects on fibrosis in vitro and in vivo. In this study, we found that the expression of heat shock protein 47 was significantly increased in patients with Schistosoma-induced fibrosis, as well as in rodent models. Immunohistochemistry revealed heat shock protein 47-positive cells were found in the periphery of egg granulomas. Administration of heat shock protein 47-targeted short hairpin (sh)RNA remarkably reduced heat shock protein 47 expression and collagen deposition in NIH3T3 cells and liver tissue of S. japonicum-infected mice. Life-table analysis revealed a dose-dependent prolongation of survival rates with the treatment of heat shock protein 47-shRNA in murine fibrosis models. Moreover, serum alanine aminotransferase and aspartate transaminase activity, splenomegaly, spleen weight index and portal hypertension were also measured, which showed improvement with the anti-fibrosis treatment. The fibrosis-related parameters assessed were expressions of Col1a1, Col3a1, TGF-β1, CTGF, IL-13, IL-17, MMP-9, TIMP-1 and PAI-1 in the liver. This study demonstrated that heat shock protein 47-targeted shRNA directly reduced collagen production of mouse liver fibrosis associated with S. japonicum. We conclude that heat shock protein 47 plays an essential role in S. japonicum-induced hepatic fibrosis in mice and may be a potential target for ameliorating the hepatic fibrosis caused by this parasite.

MicroRNA-29b-3p prevents Schistosoma japonicum-induced liver fibrosis by targeting COL1A1 and COL3A1†

Schistosomiasis is one of the worlds major public health problems in terms of morbidity and mortality, causing granulomatous inflammation and cumulative fibrosis. This study explored in vivo and vitro effects of miR‐29b‐3p in granulomatous liver fibrosis by targeting COL1A1 and COL3A1 in Schistosoma japonicum infection. Thirty male Balb/c mice were assigned to normal control and model (percutaneous infection of cercariae of S. japonicum) groups. NIH‐3T3 mouse embryonic fibroblasts were designated into blank, NC, miR‐29b‐3p mimic, TGF‐β1, TGF‐β1 + NC, and TGF‐β1 + miR‐29b‐3p mimic groups. HE and Masson staining were employed to observe the pathological changes and collagenous fibrosis. The expression of α‐SMA, COL1A1, COL3A1, TIMP‐1 was determined by immunohistochemistry. The RT‐qPCR, Western blotting and immunofluorescence staining were conducted to determine expression of miR‐29b‐3p, COL1A1, and COL3A1. CCK‐8 assay and flow cytometry were performed to evaluate viability and apoptosis. The relative expression of miR‐29b‐3p decreased in the model group. The model group showed marked fibrosis in liver tissues. The expression of α‐SMA, COL1A1, COL3A1, TIMP‐1 was higher in the model group than that in the normal control group. Dual luciferase reporter gene assay revealed that miR‐29b‐3p directly targeted COL1A1 and COL3A1. Compared with the blank, NC, TGF‐β1 and TGF‐β1 + NC groups, the miR‐29b‐3p mimic group exhibited up‐regulated expression of miR‐29b‐3p and MMP‐9 but down‐regulated expression of TIMP‐1, HSP47, α‐SMA, COL1A1, and COL3A1; while lower cell viability but higher apoptosis rate showed. It indicated that miR‐29b‐3p prevents S. japonicum‐induced liver fibrosis by inhibiting COL1A1 and COL3A1.

Effect of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis

The therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis. Fifty Kunming mice were randomly divided into 5 groups: normal control group, infection control group, anluohuaxian tablet-treated group, γ-IFN-treated group and combined treatment (anluohuaian tablet+γ-IFN) group. Pathologic changes in liver, including hepatic pigmentation and the size of schistosomal egg granuloma, were observed by HE staining after treatment for 8 weeks. The expression of the type I and collagen III, and TIMP-1 was detected by immunohistochemistry. TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR. Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group. The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months. Before and after treatment, the changes of symptoms and signs, liver function, serum liver fibrosis indexes and imaging indexes were observed. The results showed that as compared with infection control group, all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation (P<0.05). Furthermore, the expression of collagen I and III, TIMP-1, and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups (P<0.05). In the clinical observation, the serum liver fibrosis indexes, the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group (P<0.05). It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function, alleviate liver fibrosis, and could be used as a choice in treating patients with schiatosomal liver fibrosis.SummaryThe therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis. Fifty Kunming mice were randomly divided into 5 groups: normal control group, infection control group, anluohuaxian tablet-treated group, γ-IFN-treated group and combined treatment (anluohuaian tablet+γ-IFN) group. Pathologic changes in liver, including hepatic pigmentation and the size of schistosomal egg granuloma, were observed by HE staining after treatment for 8 weeks. The expression of the type I and collagen III, and TIMP-1 was detected by immunohistochemistry. TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR. Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group. The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months. Before and after treatment, the changes of symptoms and signs, liver function, serum liver fibrosis indexes and imaging indexes were observed. The results showed that as compared with infection control group, all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation (P<0.05). Furthermore, the expression of collagen I and III, TIMP-1, and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups (P<0.05). In the clinical observation, the serum liver fibrosis indexes, the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group (P<0.05). It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function, alleviate liver fibrosis, and could be used as a choice in treating patients with schiatosomal liver fibrosis.

IL-33 protects murine viral fulminant hepatitis by targeting coagulation hallmark protein FGL2/fibroleukin expression.

HighlightsMHV‐3 infection causes IL‐33 levels increasing and developing of fulminant hepatitis.IL‐33 treatment leads to attenuation of the disease and remarkable reduction of FGL2.In vitro IL‐33 administration prevents FGL2 secretion by macrophages.IL‐33 had marked effects on treating viral fulminant hepatitis by targeting FGL2. Abstract Fulminant hepatitis (FH) is characterized by rapid liver failure and high mortality. The pathogenesis of viral FH includes virus‐induced immune activation, inflammation, and subsequent hepatic apoptosis and necrosis. However, the mechanisms that underlie FH progression are unclear. IL‐33 is a member of the IL‐1‐related cytokines, considered to be an “alarmin” that participates in various diseases, but its precise role in the coagulation of FH is not very clear. In our study, we found that IL‐33 is significantly elevated in mice infected with murine hepatitis virus strain 3 (MHV‐3). This is accompanied by an increase in pro‐coagulant fibrinogen‐like protein 2 (FGL2) in the liver. Previous studies have suggested that an increase in FGL2 is diagnostic of FH and liver necrosis, and animals with no FGL2 had better survivorship during FH. Our studies showed that IL‐33 administration in a MHV‐3 infection promoted survival during FH, with a significant reduction in FGL2 expression and liver inflammation. In vitro IL‐33 treatment abrogated MHV‐3 and IFN‐&ggr; induced FGL2 expression in RAW264.7 and THP‐1 cells, respectively. In conclusion, our research suggests that IL‐33 protects against viral fulminant hepatitis in mice by antagonizing expression of the pro‐coagulant protein FGL2.

Effect of miR-182 on hepatic fibrosis induced by Schistosomiasis japonica by targeting FOXO1 through PI3K/AKT signaling pathway

The study aimed to investigate the impact of miR‐182 and FOXO1 on S. japonica‐induced hepatic fibrosis. Microarray analysis was performed to screen out differential expressed miRNAs and mRNAs. Rat hepatic fibrosis model and human hepatocellular cell line LX‐2 were used to study the effect of miR‐182 and FOXO1. qRT‐PCR and Western blot were used to detect the expression of miR‐182, FOXO1 or other fibrosis markers. The targeting relationship between FOXO1 and miR‐182 was verified by luciferase reporter assay. Immunohistochemistry or immunofluorescence staining was conducted to detect FOXO1 or α‐SMA in rat hepatic tissues. Cell viability and apoptosis were detected by MTT assay and flow cytometry. The expression of PI3K/AKT pathway‐related proteins was detected by Western blot. miR‐182 was highly expressed in liver fibrosis samples, and FOXO1 expression was negatively correlated with miR‐182 expression. After transfection of miR‐182, FOXO1 expression was down‐regulated, with the results of LX‐2 cells proliferation inhibition and apoptosis induction, as well as the aggravation of rat hepatic fibrosis. The expression of p‐AKT/AKT and p‐S6/S6 was increased, meaning that the PI3K/AKT signal pathway was activated. The results were reversed when treated with Wortmannin (PI3K inhibitor). After transfection of miR‐182 inhibitor, FOXO1 expression was up‐regulated, LX‐2 cell proliferation was inhibited, and apoptosis rate was increased. High‐expressed miR‐182 and low‐expressed FOXO1 promoted proliferation and inhibiting apoptosis on liver fibrosis cells, stimulating the development of S. japonica‐induced hepatic fibrosis through feeding back to PI3K/AKT signaling pathway.

Soluble Fgl2 restricts autoimmune hepatitis progression via suppressing Tc17 and conventional CD8+ T cell function

Autoimmune hepatitis (AIH) is an inflammatory disease caused by an aberrant immune response to hepatic self‐antigens in which regulatory T cells (Tregs) are critical for maintaining immunosupression. The soluble form of fibrinogen‐like protein 2 (sFGL2), a novel effector molecule of Treg, is rarely investigated in AIH. In the present study, we dissected the role of sFGL2 in autoimmune hepatitis and its potential mechanism underlying AIH progression.

A mixed analysis comparing nine minimally invasive surgeries for unresectable hepatocellular carcinoma patients

Hepatocellular carcinoma (HCC) is usually managed by the transcatheter arterial chemoembolization (TACE). However, this technique has been challenged since severe complications have been observed in clinical practices. As a result, clinicians have started to seek other minimally invasive surgeries with equivalent efficacy. The corresponding surgeries were assessed by the five outcomes: complete response (CR), partial response (PR), stable disease (SD), progression disease (PD) and objective response rate (ORR). Direct meta-analysis and network meta-analysis were performed and the results were represented by odds ratios (OR), 95% confidence and credential intervals. Furthermore, the value of surface under the cumulative ranking curve (SUCRA)was calculated to provide corresponding rankings.Seventeen studies were incorporated into the network meta-analysis which indicated that TACE + external-beam radiation therapy (EBRT) and drug-eluting beads (DEB) were better than TACE at controllingPD. TACE + EBRT demonstrated their advantages compared to TARE-90Y.However, network meta-analysis comparison showed no significant difference between the corresponding eight treatments with respect to CR, PR, SD and ORR. Moreover, the SUCRA suggested that TACE+EBRT were better than other treatments at treating unresectableHCC.Based on the present results of this network meta-analysis, TACE + EBRT was more effective than the other seven minimally invasive surgeries and therefore it is considered as the optimal treatment for HCC.

The expression of ETAR in liver cirrhosis and liver cancer

ABSTRACT Background: To investigate the expression of endothelin receptors in liver diseases and discuss its role in the process of liver cirrhosis and liver cancer. Research design and methods: We examined the expressions of ETAR, ETBR and α-SMA in tissue samples using western blotting analysis. Furthermore, immunofluorescence was used to locate ETAR expression in hepatic stellate cells (HSCs) and hepatic sinusoidal endothelial cells (HSECs), we calculated the percentage of positive cells and then analyzed its relation with clinical indexes. Results: According to the western blotting analysis, the expression of ETAR was high in hepatic hemangioma and liver cancer tissues and ETBR was highly expressed in cirrhosis tissues. The immunofluorescence results demonstrated that the expression of ETAR was elevated in hepatic hemangioma and liver cancer tissues. Moreover, ETAR expression was found in both HSCs and HSECs. Finally, the statistical analysis revealed that the number of positive ETAR cells was correlated with the clinical index platelets (PLT), alanine transaminase (ALT) and diameter of portal vein. Conclusion: Endothelin receptors express differently in liver cirrhosis and liver cancer tissues and play a role in hepatic diseases by affecting HSCs and HSECs.