| Background:previous studies found that by inhibiting the NF-?B signaling pathways of inflammation activity,sinomenine has a very good effect for the treatment of rheumatoid arthritis, and the nf-kappa B signaling pathway is of vital importance to the development processin of Atherosclerosis (Atherosclerosis, AS), the emergence of the foam cell is a vital inflammatory cell mark to the formation of Atherosclerosis, iconic by animal experiment and in vitro experiments found that sinomenine has the function of anti inflammation of Atherosclerosis and maintain the cholesterol metabolism balance, but the mechanism of sinomenine ease AS foam cells in the inflammatory response is unclear.This topic proposed sinomenine can also through affecting the nf-kappa B signaling pathways in the foam cell, to treat atherosclerosis disease hypothesis.Objective:Detect the biological activities and inflammatory mediators include nuclear factor kappa B,IL-1,IL-6,TNF-a with different concentration of sinomenine in vitro,to explore the possible anti-inflammatory mechanism of sinomenine in anti atherosclerosis, and provide laboratory evidence for clinical application of sinomenine anti atherosclerosis related diseases.Method:Stimulate THP-1 mononuclear cells with RPMI-1640 complete medium which containing the final concentration of 160nM 12-O-tetradecanoylphorbol 13-acetate (PMA) for 24h,then stimulate with 50mg/L low density lipoprotein (ox-LDL) for 48h to establish the foam cell model. The experiment is divided into seven groups:control group,different concentrations Sinomenine group (1?g/ml, 20?g/ml,40?g/ml,80?g/ml),5?g/ml simvastatin group and 100?g/ml dexamethasone group.Groups were given different culture medium,measure the growth activity of the foam cell after 24h,after cultivating 12 h,24h,48h,collecting the cells supernatant and cells.Detecting the secretion of inflammatory factors NF-?B,IL-1, IL-6, TNF-? by ELISA,at the same time, measuring the content of intracellular cholesterol ester,data analysis and processing applications SPSS 17.0.Results:1-. after 24h PMA stimulation and 48h ox-LDL stimulation the THP-1 monocytes will become foam cells;2?the cholesterol content of the foam cells increased, and the cholesterol content accounted for about half of the total cholesterol;3?regular training of foam cells, the cells survival rate decreases with the time goes long,and after 48h,the volume of living cell fell sharply;4?sinomenine has a certain foam cell growth inhibition betweenlug/mL-100ug/mL,when the sinomenine concentration increased,the inhibition of sinomenine enhanced;there is a significant amount-effect relationship between sinomenine and foam cells;under the 40?g/mL concentration of sinomenine intervention,we found that with the extension of time,the cell survival rate decreased, hints of sinomenine on the growth of foam cell inhibition there is a certain-effect relationship;5?after the sinomenine intervention, the content of cholesterol ester in the foam cell were cut down, and there is a significant difference with the group not give medicine (P<0.01), but no significant difference with the positive dexamethasone group;6?sinomenine can inhibit the expression of nf-kappa B in foam cell, as well as the secretion of inflammatory molecules in the downstream of nf-kappa B signaling pathways,such as IL-1, IL-6, TNF alpha, and with the increase of concentration of sinomenine, the extension of time, the inhibition enhanced, there is a certain amount-and time-effect relationship.Conclusion:1?stimulate by PMA for 24h,THP-1 monocytes can be called macrophages,then stimulate by ox-LDL for 48h,they become foam cells;2?sinomenine could by lowering the content of the nf-kappa B, as well as the nf-kappa B signaling pathways downstream related inflammatory cytokine IL-1, IL-6, TNF alpha to suppress the inflammatory response, to achieve the effect of anti inflammation of atherosclerosis;3?sinomenine could inhibit the growth activity of the ox-LDL induced THP-1 monocytes source foam cell,to induce the apoptosis of foam cell, and like the nf-kappa B,sinomenine inhibit the growth activity of the foam cell depends on the concentration of the sinomenine, its mechanism may be related to sinomenine inhibiting the activity of the nf-kappa B signaling pathway.4?sinomenine could reduce the content of cholesterol ester in foam cells, alleviate the lipid metabolism load in the cells, reach the effect of resistance to atherosclerosis, and there is no obvious difference between various concentration of sinomenine, its mechanism may be related to sinomenine inhibiting the activity of the nf-kappa B signaling pathway. |
PDF Full Text Request