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The Pro-apoptotic Effects of Berberine on Mouse Bone Marrow Derived Dendritic Cells

Author: JiaoQingQing
Tutor: WangWeiYun；ZhangWeiDong；HuZhenLin
School: Anhui Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Berberine Dendritic cells Apoptosis Rheumatoid arthritis Anti-inflammatory Immunosuppressive
CLC: R285.5
Type: Master's thesis
Year: 2010
Downloads: 59
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Abstract

Berberine (berberine, Ber) as detoxification drugs and antibiotics have been applied in clinical practice for many years. With the deepening of the research Ber initially identified berberine has anti-inflammatory and immunosuppressive effects, but relevant mechanisms been fully elucidated. Dendritic cells (dendritic cells, DC) are professional antigen-presenting cells, which can activate lymphocytes and initiate the immune response, while maintaining immune tolerance also plays a key role. Current studies suggest that DC maturation status may be an important immunoregulatory control point determines the direction of the immune response: the immune response or immune tolerance. On the other hand, DC's timely removal of preventing overreaction of the immune system is very important. DC continued survival will lead to the accumulation of a large number of DC and eventually lead to chronic lymphocyte activation and autoimmunity. Thus, DC maturation status and survival time required to ensure precise adjustment of the normal adaptive immune response, while avoiding the occurrence of autoimmune response. Rheumatoid arthritis (RA) is a kind of synovitis characterized by chronic systemic autoimmune disease, the current findings indicate that DC is caused by chronic proliferative synovitis and joint destruction process of immune response important role. For a long time, some of the classic immunosuppressants (eg corticosteroids, cyclosporine A, rapamycin and 1,25 (OH) 2D3, etc.) produced by immune play is considered primarily through inhibition of T cell activation. But the latest research shows that these drugs could inhibit the varying degrees of DC activation induces DC maturation or apoptosis. Thus, DC may be an important target for prevention and treatment of RA cells. Previous studies have shown anti-inflammatory and immunosuppressive effects Ber, while toxicity is low, but so far there has not been affected berberine on DC or RA reported. The purpose of this study proved Ber on DC as well as on RA for potential therapeutic role in order to investigate the mechanism of anti-inflammatory and immunosuppressive effects, as well as the prevention and treatment of RA Ber develop drugs for the foundation. The main contents and results are as follows: In vitro: By AnnxinV / PI staining to detect changes in nuclear morphology, mitochondrial membrane potential, reactive oxygen species, cell cycle changes of berberine on cultured mouse bone marrow-derived dendritic cells (BMDC), peritoneal macrophages, RAW264.7, Jurkat T cells as well as other types of tumor cell apoptosis. The results show that BMDC culture within the system, different doses of berberine (2-50μM) can induce DC apoptosis; BMDC growth and differentiation in the first three days to detect berberine-induced apoptosis; mature DC ( mDC) than non-mature DC (imDC) for inducing apoptosis in Ber more sensitive; while freshly isolated bone marrow cells, peritoneal macrophages, RAW264.7 and Jurkat T cells inducing apoptosis in Ber insensitive. Although berberine in some tumor cell lines showed apoptotic role, but DC than berberine these tumors more sensitive cell lines. Vivo test: the collagen Ⅱ-induced arthritis in mice (CIA) for the model to detect anti-inflammatory and immunosuppressive effects of berberine. By serum antibody levels, cytokine production, lymphocyte assay value on collagen Ⅱ mice investigated the immune response. On the spleen and lymph nodes draining DC DC cell surface markers and TUNEL staining and flow cytometry and immunohistochemistry method to detect apoptosis. The results show that the CIA model, after intraperitoneal injection of berberine rheumatoid arthritis incidence and severity of anti-collagen antibody levels Ⅱ, lymphocytes and spleen cells added IFN-γ and IL-17 were significantly lower water In addition within the spleen and lymph nodes also significantly reduced the number of DC, and the DC apoptosis rate increased significantly. These results show that: berberine selectively induce apoptosis in DC, on collagen Ⅱ-induced arthritis mouse model showed significant anti-inflammatory and immunosuppressive effects.

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