摘要
目的:观察不同相对分子质量的一年生膜荚黄芪多糖(APSⅠ)对脂多糖(LPS)诱导的RAW264.7炎症模型分泌促炎因子和抗炎因子的影响,探讨APSⅠ的抗炎作用。方法:体外培养RAW264.7细胞,APSⅠ作用于未经刺激的RAW264.7细胞及LPS(1mg·L-1)刺激后RAW264.7细胞,将细胞分为空白对照组、LPS模型组、APSⅠ组及LPS+APSⅠ不同相对分子质量组(APSⅠ-A,APSⅠ-B,APSⅠ-C),采用CCK-8法检测不同相对分子质量、不同浓度的APSⅠ组RAW264.7细胞增殖活力,硝酸还原酶法测定细胞上清液中一氧化氮(NO)的含量,酶联免疫吸附法(ELISA)检测细胞上清中肿瘤坏死因子α(TNF-α)、白细胞介素10(IL-10)的分泌量。结果:与空白对照组比较,APSⅠ单独处理组RAW264.7细胞增殖活性降低(P<0.05或P<0.01),NO及IL-10表达水平明显增加(P<0.05),TNF-α的分泌量降低(P<0.05);与LPS模型组比较,LPS+APSⅠ组RAW264.7细胞增殖活力显著升高(P<0.05或P<0.01),NO、TNF-α的表达明显降低(P<0.05或P<0.01),IL-10的分泌增加(P<0.05或P<0.01);3种不同相对分子质量APSⅠ之间比较,APSⅠ-C对NO、TNF-α的抑制更为明显,且APSⅠ-C促进IL-10分泌的作用强于APSⅠ-A及APSⅠ-B。结论:APSⅠ可以拮抗LPS所致的RAW264.7细胞炎症反应;不同相对分子质量APSⅠ的抗炎作用有差异,APSⅠ的生物活性与其相对分子质量存在一定的关联性。
Objective To investigate the effects of different molecule weight astragalus polysacharin isolated from the annual astragalus membra-neaceus(APSⅠ)on the expressions of pro-inflammatory and anti-inflammatory cytokine in lipopolysaccharides(LPS)-stimulated RAW264.7 cells,and explore the anti-inflammatory effects of APSⅠ.Methods The RAW264.7 cells were cultivated in vitro,RAW264.7 cells or LPS-induced(1 mg·L-1) RAW264.7 cells were stimulated by APS Ⅰ,then divided into control group,LPS model group,APS Ⅰ group,LPS + different molecule weight APS Ⅰgroups(APS Ⅰ-A,APSⅠ-B,APSⅠ-C).The effects of different molecule weight and different concentrations of APSⅠon the proliferation of RAW264.7 cells were examined by CCK-8 method.The content of NO in the RAW264.7 cells supernatant fluid was detected by means of nitrate reductase.The concentrations of TNF-α and IL-10 in the supernatant fluid was determined with ELISA.Results Compared with control group,the proliferation of RAW264.7 cells was inhibited significantly after treated with APS Ⅰalone(P<0.05 or P<0.01),the levels of NO and IL-10 expressions were increased apparently(P<0.05) while the secretion of TNF-α was decreased(P<0.05);Compared with LPS model group,APS Ⅰ combined with LPS significantly increased the proliferation activity of RAW264.7 cells in LPS + APS Ⅰ groups(P<0.05 or P<0.01),the levels of No and TNF-α expressions were decreased significantly(P<0.05 or P<0.01),and the secretion of IL-10 was increased notably(P<0.05 or P<0.01).The effects of APS varied by different molecular weights,the inhibition of APSⅠ-C on the expressions of NO and TNF-α was more obvious,which had a stronger promotion on the secretion of IL-10 than APSⅠ-A and APS-B.Conclusion APSⅠhas anti-inflammatory effect on the LPS-stimulated RAW264.7 cells,the anti-inflammatory activation of APSⅠvaries by different molecule weights.APSⅠwith small molecule weight has more significant activation.So the biological activity of APSⅠmay be associated with molecule weight.
出处
《吉林大学学报(医学版)》
CAS
CSCD
北大核心
2011年第6期1051-1056,共6页
Journal of Jilin University:Medicine Edition
基金
长白山生物资源与功能分子教育部重点实验室资助课题[长生实合字(2008)第05号]