Angiotensin Ⅱ Type 2 Receptor Gene Transfection Inhibit Maturation of Bone Marrow Derived Dendritic Cells from Mice in Vitro
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    Abstract:

    Aim To investigate the influence of angiotensin Ⅱ type 2 receptor (AT2R) gene transfection on the maturation and some immune functions of bone marrow derived dendritic cells (BMDC) from mice and explore the mechanism of AT2R-induced immunologic function in atherogenesis. Methods The cell suspension was made from C57BL/6J mice marrow,purified and differentiated to BMDC. First the BMDC were transfected with pAdCMV/AT2R or control virus pAd-GFP,containing green fluorescent protein reporter gene Then were stimulated by 100 μg/L lipopolysaccharide (LPS) into mature DC In the same time were added to antagonist of AT2R as pAdCMV/AT2R+PD123319 group PBS as negative control and LPS as positive control. The expressions of AT2R in DC were evaluated by RT-PCR,immunofluorescence staining and confocal microscope. The CD86 and MHCⅡ expressing rates were detected by fluorescence-activated cell sorting (FACS). Liquid scintillation counting(LSC) was used in mixed lymphocyte reactions (MLR) to reflect the ability of BMDC to stimulating homologous T cells proliferation. Cytokines IL-12 and IFN-γ were detected by ELISA.Results After pAdCMV/AT2R was transduced into BMDC,the expressions of AT2R mRNA significantly increased at 48 hours and 72 hours,green fluorescence localized to cell nuclei and plasm,red fluorescence labling AT2R was present in cell plasm. Compared with PBS negative control,the expressions of CD86,MHCⅡ was significantly up-regulated in pAdCMV/AT2R group and pAdCMV/AT2R+PD123319 group (P<0.01),the stimulating capacity of BMDC obviously enhanced (P<0.01),levels of IFN-γ and IL-12 in the supernatant increased(P<0.01) But compared with LPS positive control,the changes of pAdCMV/AT2R group were significantly decreased(P<0.01 or P<0.05), the changes of pAdCMV/AT2R+PD123319 group were not significantly different(P>0.05). Conclusion BMDC can express AT2R stably in vitro,AT2R can inhibit the maturation and immunologic action of BMDC and antagonist of AT2R can hold up the effect,so AT2R may induce inhibition of atherosclerosis.

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TANG Bing, SU Xiao-Hua, CHEN Jing-Song, LI Gang, YANG Da-Chun, YANG Yong-Jian, ZHU Jun, LI Xing-Ke, and LI De. Angiotensin Ⅱ Type 2 Receptor Gene Transfection Inhibit Maturation of Bone Marrow Derived Dendritic Cells from Mice in Vitro[J]. Editorial Office of Chinese Journal of Arteriosclerosis,2015,23(09):881-886.

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  • Received:November 14,2014
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  • Online: July 21,2015
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