红毛五加多糖单一组分AHP-Ⅲ对巨噬细胞的免疫调节作用及其机制

Immunomodulatory effect and its mechanism of Acanthopanax giraldii Hams polysaccharideⅢon macrophages

探讨红毛五加多糖(Acanthopanax giraldii Hams polysaccharide)单一组分AHP-Ⅲ(Acanthopanax giraldii Hams polysaccharideⅢ)对小鼠巨噬细胞RAW 264.7的激活作用及机制。不同浓度AHP-Ⅲ作用RAW 264.7细胞,中性红试验检测细胞吞噬能力;ELISA和Griess法检测其IL-6、TNF-α和NO的释放量;RT-qPCR检测iNOS、TNF-α和IL-6 mRNA相对表达水平;Western blot检测NF-κB信号通路相关蛋白磷酸化水平。在实验浓度范围内,AHP-Ⅲ可显著增强RAW 264.7细胞的吞噬能力(P<0.05);促进RAW 264.7分泌NO、TNF-α和IL-6(P<0.05或P<0.001);并显著增加RAW 264.7细胞中IL-6、TNF-α和iNOS mRNA的表达量,呈剂量依赖性;Western blot结果表明,AHP-Ⅲ作用RAW 264.7细胞后,NF-κB中的p65、IKKβ、IκBα磷酸化水平明显升高。结果显示红毛五加多糖AHP-Ⅲ对小鼠巨噬细胞RAW 264.7具有显著激活作用。

To study the activation and mechanism of Acanthopanax giraldii Hams polysaccharideⅢ(AHP-Ⅲ)on mouse macrophage RAW 264.7.After RAW 264.7 macrophages were stimulated with the different concentrations of AHP-Ⅲ,the phagocytic ability was conducted by neutral red method.The release capacity of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and NO under different concentrations of AHP-Ⅲstimulation were tested by the ELISA and the Griess,and the transcription level of inducible NO synthase(iNOS),TNF-αand IL-6 was determined by RT-qPCR,then were detected by Western blot phosphorylation of NF-κB signaling pathway related proteins.Compared with normal control group,AHP-Ⅲcan significantly improve the phagocytic ability of RAW 264.7(P<0.05)within the test concentration range.Also,it can promote RAW 264.7 to secrete NO,TNF-αand IL-6(P<0.05 or P<0.001).According to the result of RT-qPCR,in a dose-dependent manner,AHP-Ⅲcan significantly improve the expression level of IL-6,TNF-αand iNOS.Western blot suggested that after stimulation of AHP-Ⅲ,the phosphorylation level of p65,IKKβand IκBαin NF-κB were significantly improved.The results showed that AHP-Ⅲcould significantly activate RAW 264.7 mouse macrophage.