摘要
目的探讨党参多糖对胃癌细胞AGS增殖、凋亡和炎症因子表达的影响和机制。方法采用CCK-8实验、流式细胞术评估党参多糖(10、20、40μmol/L)对胃癌细胞AGS活力和凋亡的影响。RT-qPCR检测miR-361-5p表达;ELISA法检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-1β分泌。蛋白质免疫印记法检测TLR4和磷酸化核因子(NF)-κBp65(p-p65)和磷酸化NF-κB抑制蛋白-α(p-IκBα)蛋白的表达。将miR-361-5p模拟物、miR-361-5p抑制物分别转染AGS细胞,经40μmol/L党参多糖处理后,用上述方法检测AGS细胞活力、凋亡率、Toll样受体4(TLR4)、p-p65和p-IκBα蛋白表达以及TNF-α、IL-6、IL-1β分泌。结果10、20、40μmol/L党参多糖显著降低AGS细胞活力(P<0.05),增加细胞凋亡率(P<0.05),抑制TNF-α、IL-6、IL-1β分泌(P<0.05),并上调miR-361-5p表达水平(P<0.05)。40μmol/L党参多糖明显降低TLR4、p-p65和p-IκBα蛋白表达水平(P<0.05)。过表达miR-361-5p联合40μmol/L党参多糖能够降低AGS细胞活力(P<0.05),增加细胞凋亡率(P<0.05),抑制TNF-α、IL-6、IL-1β分泌(P<0.05),抑制TLR4、p-p65和p-IκBα蛋白表达(P<0.05)。抑制miR-361-5p后显著减弱40μmol/L党参多糖对AGS细胞活力、凋亡率、促炎因子分泌以及TLR4、p-p65和p-IκBα蛋白表达的影响(P<0.05)。结论党参多糖可有效抑制胃癌AGS细胞增殖,诱导细胞凋亡,抑制促炎因子分泌,其机制可能与调控miR-361-5p/TLR4/NF-κB通路有关。
This study was designed to investigate the effect of Codonopsis pilosum polysaccharide on the proliferation,apoptosis and inflammatory factor expression of gastric cancer cells AGS and its mechanism.CCK-8assay and flow cytometry were applied to evaluate the effects of Codonopsis pilosum polysaccharides(10,20,40μmol/L)on the viability and apoptosis of AGS cells;RT-qPCR was used to detect the expression of miR-361-5p;corresponding kits were selected to detect the secretion of tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-1β;Western blotting was used to detect the protein expression of TLR4,phosphorylated nuclear factor(NF-κB p65,p-p65)and phosphorylated NF-κB inhibitory protein-α(p-IκBα).After the transfection of miR-361-5p mimics or miR-361-5p inhibitor,the AGS cells were treated with 40μmol/L Codonopsis pilosum polysaccharides,and then the indicators mentioned above were measured.Data showed that Codonopsis pilosum polysaccharides(10,20,40μmol/L)significantly reduced the viability of AGS cells(P<0.05),increased cell apoptosis rate(P<0.05),inhibited the secretion of TNF-α,IL-6,and IL-1β(P<0.05),and up-regulated the expression level of miR-361-5p(P<0.05).Furthermore,Codonopsis pilosum polysaccharide of40μmol/L significantly reduced the expression levels ofTLR4,p-p65 and p-IκBαproteins(P<0.05).mi R-361-5 p overexpression combined with 40μmol/L Codonopsis pilosum could reduce the viability of AGS cells(P<0.05),increased cell apoptosis rate(P<0.05),and inhibited thesecretion of TNF-α,IL-6,and IL-1β(P<0.05),inhibited the expression of TLR4,p-p65 and p-IκBαproteins(P<0.05).mi R-361-5 p inhibition significantly reduced the effects of 40μmol/L Codonopsis pilosum polysaccharide oncell viability,apoptosis rate,pro-inflammatory factors secretion,as well as TLR4,p-p65 and p-IκBαproteinexpression of AGS cells(P<0.05).In conclusion,Codonopsis pilosum polysaccharide can effectively inhibit gastriccancer AGS cells proliferation,induce cell apoptosis,and inhibit pro-inflammatory factors secretion,throughregulating miR-361-5 p/TLR4/NF-κB pathway.
作者
杨自豪
周亨露
YANG Zihao;ZHOU Henglu(Department of pharmacy,the Second Affiliated Hospital of PLA Army Medical University,Chongqing 400037,China;Department of pharmacy,Chongqing Jiulongpo District People's Hospital,Chongqing 400050,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2022年第4期347-353,共7页
Immunological Journal
