Alum Borneol Nanoemulsion Promotes M2 Type Polarization of Macrophages and Inhibits the Expression of Inflammatory Factors
Objective To observe the effects of alum borneol nanoemulsion on the secretion of inflammatory factors by lipopolysac-charide(LPS)induced mononuclear phagocyte(RAW264.7)in mice.Methods Using LPS to induce RAW264.7 as an M1 pheno-type,the changes of cell morphological were observed under microscope.The effects of different concentrations of alum borneol nanoemul-sion on RAW264.7 cell viability was detected by methylthiazolyldiphenyl-tetrazolium bromide(MTT)colorimetry.The relative mRNA expression of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and interleukin-10(IL-10)were detectd by real-time quantitative polymerase chain reaction(RT-PCR).The positive expression rates of M1/M2 polarized surface markers CD68 and CD206 were measured by flow cytometry The contents of IL-6,TNF-α and IL-10 in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Results The morphology of RAW264.7 cells changed obviously when cultured with LPS and alum bor-neol nanoemulsion in vitro.Low,medium,and high concentrations(8.15,16.30,32.60mg/ml)of alum borneol nanoemulsion had no toxic effects on RAW264.7.Compared with the blank group without any treatment,the expression of CD68 was higher(P＜0.001),the expression of CD206 was lower(P＞0.05)in the model group after 24hours of LPS stimulation of RAW264.7 cells with good growth,in-dicating that M1 macrophage modeling was successfully.After drug intervention,compared with the model group,the expression of CD68membrane surface molecules was lower than before(P＜0.001),and CD206 was higher than before(P＜0.001),indicating that alum borneol nanoemulsion could increase the expression of M2macrophage membrane protein.Compared with the model group(LPS group),alum borneol nanoemulsion could inhibit the secretion of TNF-α and IL-6 in LPS-induced macrophage inflammation model(P＜0.01,P＜0.05),but promoted the secretion of IL-10(P＜0.001).Compared with the model group(LPS group),the results of RT-qPCR showed that alum borneol nanoemulsion significantly inhibited the expression of TNF-α and IL-6mRNA(P＜0.01,P＜0.05),but promoted the expression of IL-10mRNA(P＜0.01).Conclusion The alum borneol nanoemulsionhas has good anti-in-flammatory activity on LPS-induced inflammatory responses in RAW264.7 cells,which may be achieved by inhibiting the secretion and expression of inflammatory cytokines.
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