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.
NETL
NSTL
万方数据
