Junfa Yang

and 9 more

Background and Purpose: Extracellular matrix (ECM) is mainly derived from activated hepatic stellate cells (HSC), and its excessive deposition is one of the characteristics of liver fibrosis. Monomer derivative of paeoniflorin (MDP) inhibits inflammatory responses. However, the role and fundamental mechanism of MDP in liver fibrosis was still unclear. Experimental Approach: The effect of MDP was evaluated on CCl4-induced C57BL/6J mice and TGF-β1-induced LX-2 cells. The level of SA-β-Gal was detected by SA-β-Gal kits. The cell cycle was evaluated by flow cytometry. The expression of p16, p21, α-SMA and Col. I proteins were analyzed by qRT-PCR, Western blots and immunofluorescence staining and IHC staining. The pathological changes of liver tissue were evaluated by histological analysis. Key Results: We demonstrated that MDP inhibited the progression of liver fibrosis in vivo and in vitro, concomitant with the elevated expression of Ago2, miR-708 and p53 as well as the downregulated expression of ZEB1. MDP could combined with Ago2. Upregulation of miR-708 and p53 and downregulation of ZEB1 increased the number of SA-β-Gal-positive HSCs and the expression levels of p16 and p21 as well as decreased the expression of α-SMA and Col. I in activated HSCs. Meanwhile, miR-708 directly targeted ZEB1, thus inhibiting the mRNA of ZEB1. ZEB1 could bind to the E‐box of p53 promoter and restrain its promoter activity as well as thus block the expression of p53. Conclusion and Implications: MDP could induce senescence of activated HSCs via regulating Ago2/miR-708/ZEB1/p53 aixs and may be applied to treat liver fibr

Xianzheng Zhang

and 16 more

Li Xu

and 11 more

Abstract Purpose. To investigate the mechanisms of macrophage pyroptosis mediated by TLR4/NLRP3/GSDMD signaling pathway in adjuvant arthritis (AA) rats and the role of Paeoniflorin-6′-O-benzene sulfonate (CP-25). Experimental Approach. AA model was induced in Wistar rats via complete Freund’s adjuvant. Normal group, AA model group, CP-25 (50 mg/kg) group and MTX (0.5 mg/kg) group were included in this experiment. The co-expression of TLR4 and NLRP3 and membrane expression of GSDMD and NLRP3 in macrophages were detected by immunofluorescence assay. The expression of TLR4, the ratio of macrophage pyroptosis and M1/2-type macrophages were detected by Flow Cytomery. Cell morphology was observed by scanning electron microscopy. The levels of IL-18 and IL-1β cytokines in plasma and supernatant of cultured macrophage were detected by ELISA. The expression of TLR4, MyD88, NLRP3, Caspase-1, ASC and GSDMD in macrophages was detected by Western Blot. Key Results. Macrophage pyroptosis was found in AA rats; CP-25 has a therapeutical effect on AA rats by improving the joint inflammation and reducing the pathological process of the joints of AA rats; CP-25 can inhibit the pyroptosis of macrophages by down-regulate the expression of TLR4, MyD88, NLRP3, Caspase-1, ASC and GSDMD of macrophages in vivo; CP-25 inhibits LPS and ATP-induced macrophages pyroptosis by inhibiting the activation of TLR4/NLRP3/GSDMD signaling pathway in vitro. Conclusion and Implications. Macrophage pyroptosis was mediated through TLR4/NLRP3/GSDMD signaling pathway, and CP-25 can regulate macrophage pyroptosis by inhibiting TLR4/NLRP3/GSDMD signaling pathway, thereby improving synovitis in AA rats.

Xiao xi Hu

and 7 more

Rheumatoid arthritis(RA) is a chronic systemic autoimmunediseasecharacterized by synovitis and the destruction of small joints.Emerging evidence had shown thatthe stimulation of immunoglobulin D (IgD) induced T cell activation which may contribute to diseases pathogenesis in RA.In this study we demonstrated that IgD could induce the activation of T cells through affecting IgDR-Lck-ZAP70- PI3K-NF-κB signaling, IgD-induced CD4+T cells promoted the proliferation of CD19+B cells in RA patients. IgD-Fc-Ig fusion protein (composed of human IgD Fc domain and IgG1 Fc domain, specifically blocks the IgD-IgDR pathway)inhibited the co-expression of IgDR and p-Lck and the expressions of p-Lck, p-ZAP70, p-PI3K on CD4+T cells, and decreased NF-κB nuclear translocation in Jurkat cells. Meanwhile, IgD-Fc-Ig down-regulated the protein expressions of CD40L on CD4+T cells and CD40, CD86 on CD19+B cells in RA patients and healthy controls. It also decreased the protein expressions of CD40L on CD4+T cellsand CD40 on CD19+B cells from spleens of CIA mice and reduced IL-17A level in mouse serum. Moreover, in vivo, IgD-Fc-Ig administration dose-dependently down-regulated the protein expressions of CD40, CD40L and IgD in spleens from CIA mice. IgD-Fc-Ig restrains the activations of T cells through inhibiting IgD-IgDR-Lck-ZAP70- PI3K-NF-κB signaling, thus inhibiting the activation of B cells.Our data provides experimental evidence for application prospect of IgD-Fc-Ig as a highly selective targeting T cell treatment for RA.

Xianzheng Zhang

and 15 more

Abstract Background and purpose: To investigate the effect of hIgDFc-Ig (DG), a new biological agent targets competition for binding to IgD receptors, on collagen-induced arthritis and its potential mechanism in regulating B cell antigen-receptor signaling pathway. Experimental approach: DBA1 mice were used to establish collagen-induced arthritis model, three doses DG were administered by intraperitoneal injection. Clinical assessment of CIA, histopathological examination, flow cytometry, western blotting, immunofluorescence staining, protein chips and so on were used to evaluated therapeutic effects. The competitive effects on BCR-NF-κB signaling pathway were also evaluated in Daudi cell lines in vitro. Key results: We found that DG has a obvious therapeutic effect on CIA mice. DG relieved the clinical assessment of CIA mice and improved the pathology of joints and spleen. In addition, regulated B cell subsets in the PBMC and spleen of CIA mice, and decreased level of immunoglobulins. DG can inhibit the over-activation of BCR signal by increasing p-Lyn level, Co-treatment with DG (0.1-10 μg·ml-1) dose-dependently down-regulated the BCR signaling and decreased the iteraction between Syk and Btk stimulated by IgD in Daudi cell. Conclusion and Implications: DG may play a therapeutic role in CIA mice by regulating BCR-Lyn-Syk-NF-KB signaling pathway, and may be a new promising biological agent for rheumatoid arthritis. Key words: hIgDFc-Ig; B cell antigen receptor; Collagen-induced arthritis; NF-κB signaling pathway