不同相对分子质量一年生膜荚黄芪多糖对RAW264.7细胞炎症相关因子表达的影响

Effects of different molecule weight astragalus polysacharin isolated from annual astragalus membra-neaceus on experssions of inflammatory cytokines in RAW264.7 cells

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.