中药新药与临床药理2025，Vol.36Issue(9)：1458-1470,13.DOI:10.19378/j.issn.1003-9783.2025.09.006
枸杞多糖通过MAP3K3通路降低PD-L1表达抗肺腺癌作用及机制
The Mechanism and Anti-Lung Adenocarcinoma Effects of Lycium barbarum Polysaccharide Suppressing PD-L1 Expression via the MAP3K3 Pathway
摘要
Abstract
Objective:This study aims to investigate the anti-lung adenocarcinoma effects and underlying mechanisms of Lycium barbarum polysaccharide(LBP)in suppressing PD-L1 expression via the MAP3K3 pathway.Methods(1)In vivo experiments:A subcutaneous lung adenocarcinoma xenograft model was established in mice to evaluate the effects of low-,medium-,and high-dose LBP(25,50,and 100 mg·kg-1)on tumor growth and immune organ indices.(2)In vitro experiments:H1299 and A549 cells were treated with LBP(0.25,0.5,and 1 mg·mL-1)for 48 hours.Cell proliferation was assessed using CCK-8 and colony formation assays,while PD-L1 protein and mRNA expression levels were measured via Western Blot and RT-qPCR.A co-culture system of activated peripheral blood mononuclear cells(PBMCs)from healthy donors and lung adenocarcinoma cells was established to examine the immunomodulatory effects of LBP on PBMC-mediated tumor cell killing.Transcriptome sequencing was performed on H1299 cells treated with 1 mg·mL-1 LBP to identify differentially expressed genes(DEGs),followed by KEGG enrichment analysis.Western Blot and RT-qPCR were used to validate MAP3K3 expression changes in lung adenocarcinoma cells post-LBP treatment.(3)Rescue experiments:A MAP3K3-overexpressing A549 cell line(A549 MAP3K3 OE)was constructed.PD-L1 protein expression was assessed by Western Blot after LBP treatment,and the impact of MAP3K3 overexpression and LBP intervention on PBMC-mediated cytotoxicity was evaluated.Results(1)Compared with the control group,LBP treatment significantly reduced tumor weight(P＜0.05,P＜0.01,P＜0.001)and decreased tumor cell density while increasing stromal lymphocyte infiltration.The spleen and thymus indices were elevated in LBP-treated mice(P＜0.05).(2)LBP did not significantly alter the proliferation of lung adenocarcinoma cells after 48 h ours of treatment but downregulated PD-L1 protein and mRNA expression(P＜0.05,P＜0.01,P＜0.001).LBP enhanced the cytotoxic activity of activated PBMCs against lung adenocarcinoma cells(P＜0.05,P＜0.01,P＜0.001).Transcriptome sequencing revealed DEGs primarily enriched in the MAPK pathway.MAP3K3 protein and mRNA expression levels were significantly downregulated in LBP-treated cells(P＜0.05,P＜0.01,P＜0.001).(3)A549 MAP3K3 OE cells exhibited elevated PD-L1 expression(P＜0.01)and resistance to PBMC-mediated killing(P＜0.001).However,LBP treatment partially restored PBMC cytotoxicity against A549 MAP3K3 OE cells(P＜0.01,P＜0.001).Conclusion LBP suppresses lung adenocarcinoma progression by downregulating MAP3K3 and PD-L1 expression,thereby enhancing PBMC-mediated immune cytotoxicity in vitro.This mechanism may contribute to the anti-tumor effects of LBP against lung adenocarcinoma.关键词
枸杞多糖/MAP3K3通路/PD-L1/肺腺癌/免疫杀伤/小鼠/H1299细胞/A549细胞/HBE细胞/Lewis细胞Key words
Lycium barbarum polysaccharide/MAP3K3 pathway/PD-L1/lung adenocarcinoma/immune cytotoxicity/mice/H1299 cells/A549 cells/HBE cells/Lewis cells分类
医药卫生引用本文复制引用
陈思柔,陈文雅,陈静芹,黄秋茵,李龙娟,张韧,何彦丽..枸杞多糖通过MAP3K3通路降低PD-L1表达抗肺腺癌作用及机制[J].中药新药与临床药理,2025,36(9):1458-1470,13.基金项目
国家自然科学基金资助项目(81873154). （81873154）
