摘要
目的研究注射用七叶皂苷钠(sodiumaescinate injection,SAI)及效应成分混合物SAK(SA-A和SA-C原比例混合)的体外抗炎作用及机制。方法体外培养小鼠单核巨噬细胞白血病细胞(RAW264.7),用脂多糖(lipopolysaccharides,LPS)诱导体外炎症模型。设正常对照组、模型对照组及SAI和SAK给药组(0.4~40μg/mL)。以细胞活力、NO释放、细胞内钙离子浓度、ROS水平和iNOS、COX-2mRNA表达等作为评价指标,探讨SAI和SAK对各项指标的影响。结果LPS可诱导RAW264.7细胞上清液中的NO释放,使胞内的Ca^(2+)浓度和ROS增加。而10μg/mL的SAI和SAK可显著抑制上述指标,且10μg/mL的SAK作用强于SAI;同时,SAI和SAK对LPS诱导RAW264.7细胞iNOS、COX-2mRNA的表达上调具有抑制作用。结论SAI和SAK均可以抑制LPS诱导的RAW264.7细胞炎性反应,其抗炎机制与抑制iNOS、COX-2mRNA的表达有关,且SAK与SAI比较,其在去除SA-B和SA-D的毒性基础上抗炎活性更强,对于制药生产具有一定借鉴意义。
Objective This study was designed to evaluate the anti-inflammatory effect and mechanisms of SAI and SAK(SA-A and SA-C)in vitro.MethodsRAW264.7 cells were stimulated by lipopolysaccharide(LPS)to establish an in vitro inflammatory model.Cells were divided into groups as follows:normal control group,model control group,SAI and SAK groups(0.4-40μg·mL^(-1)).Cell viability was measured.The nitric oxide(NO)release,intracellular calcium concentration and ROS level were measured.The expression levels of iNOS and COX-2 mRNA were detected.The effect of SAI and SAK on the above indicators were evaluated.ResultsLPS can induce NO release in the supernatant of RAW264.7 cells,increasing the Ca^(2+)+concentration and the ROS level in cells.A concentration of 10μg·mL^(-1) of SAI or SAK significantly inhibited the above indicators,and 10μg·mL^(-1) of SAK was more effective than that of SAI.Also,SAI and SAK inhibited LPSinduced up-regulation of iNOS and COX-2 mRNA expressions in RAW264.7 cells.Conclusion Both of SAI and SAK can inhibit the LPS-induced inflammatory response in RAW264.7 cells.The anti-inflammatory mechanism is related to the inhibition of iNOS and COX-2mRNA expressions.Moreover,SAK has more anti-inflammatory activity after removing the toxic SA-B and SA-D contents when compared with SAI.This study has certain significance for the drug production.
作者
黄先菊
申李丹
王大贵
柏远
李竣
武宙阳
HUANG Xianju;SHEN Lidan;WANG Dagui;BO Yuan;LI Jun;WU Zhouyang(School of Pharmacy,Central South Minzu University,Wuhan,430074,China;Department of Anesthesiology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,430030,China)
出处
《医学分子生物学杂志》
CAS
2022年第5期388-393,共6页
Journal of Medical Molecular Biology
基金
重点研发计划中医药现代化研究重点专项(No.SQ2017YFC17010086-4)
2018年度湖北省技术创新专项重大项目(No.2018ACA126)
湖北省支持企业技术创新发展项目(科技型中小企业)(No.2021BLB174)。
