蒙药清咽六味散对巨噬细胞的免疫调节作用研究OA

Objective It is to investigate the effect of Mongolian medicine Qingyanliuwei powder on(QP)macrophage proliferation,migration and its immunomodulatory effects under inflammation status.Methods RAW 264.7 cells at loga-rithmic growth phase were taken and their proliferation rates after treatment with different concentrations of QP were as-sessed using the CCK-8 assay to screen out the optimal concentration for subsequent experiments.3 groups were established in the scratch experiment:a control group,a 25 μg/mL QP group,and a 50 μg/mL QP group,the scratch healing rates of cells in each group were calculated.4 groups were established in the lipopolysaccharide(LPS)stimulation experiment:the control group was treated with culture medium containing a small amount of PBS,the LPS group was treated with culture me-dium containing LPS at a final concentration of 100 ng/mL,the LPS+25 μg/mL QP group was treated with culture medium containing LPS at a final concentration of 100 ng/mL and QP at a concentration of 25 μg/mL,and the LPS+50 μg/mL QP group was treated with medium containing LPS at a final concentration of 100 ng/mL and QP at a concentration of 50 μg/mL.After 24 hours of culture,the mRNA expressions of macrophage polarization markers including inducible nitric oxide syn-thase(iNOS),arginase-1(Arg-1)and the inflammatory factors interleukin(IL)-1β and IL-6 in the cells were detected by qRT-PCR,the positive expression of iNOS in the cells was detected by immunofluorescence staining,and the levels of IL-1β and IL-6 in the cells were measured by ELISA.Results The cell proliferation rates in the 12.5 μg/mL,25 μg/mL,and 50 μg/mL QP groups were all significantly higher than that in the control group(all P<0.05);the cell scratch heal-ing rate in the 50 μg/mL QP group was significantly higher than that in the control group(P<0.05).Compared with the control group,the relative mRNA expressions of iNOS,IL-1β and IL-6,as well as the levels of IL-1β and IL-6 in the cells of the LPS group were significantly increased(all P<0.05),while the relative mRNA expression of Arg-1 was significantly reduced(P<0.05),and the fluorescence intensity of iNOS-positive expression in the cells was significantly enhanced.Compared with the LPS group,the relative mRNA expressions of iNOS and IL-1β in the cells of the LPS+25 μg/mL QP group were both significantly reduced(both P<0.05);the relative mRNA expressions of iNOS,IL-1β and IL-6,as well as the levels of IL-1β and IL-6 in the cells of the LPS+50 μg/mL QP group were significantly reduced(all P<0.05),while the relative mRNA expression of Arg-1 was significantly increased(P<0.05);the fluorescence intensities of iNOS-positive expressions were both significantly reduced in the LPS+25 μg/mL QP group and LPS+50 μg/mL QP group,and the relative mRNA expressions of iNOS and IL-1β in the cells of the LPS+50 μg/mL QP group were significantly lower than those in the cells of the LPS+25 μg/mL QP group(both P<0.05).Conclusion Qingyanliuwei powder can enhance macrophage proliferation and migration,and inhibit polarization towards the M1 phenotype and the secretion of pro-inflam-matory cytokines under inflammatory status.