湖南中医药大学学报2025，Vol.45Issue(5)：825-835,11.DOI:10.3969/j.issn.1674-070X.2025.05.007
熊果酸对非小细胞肺癌H1975细胞的体内外抑制作用研究
In vitro and in vivo inhibitory effects of ursolic acid on non-small cell lung cancer H1975 cells
摘要
Abstract
Objective To investigate the in vitro and in vivo inhibitory effects of ursolic acid(UA)on non-small cell lung cancer(NSCLC)H1975 cells,and explore its effects on endogenous metabolites in tumor-xenografted nude mice.Methods H1975 cells were used as the research object,and the optimal concentration of UA(0-75 μg/mL)for cell treatment was screened using the MTT assay.Cells were divided into control group,and low-,medium-,and high-dose UA(9.27 μmol/L,18.53 μmol/L,27.8 μmol/L)groups and all groups were treated for 48 hours.Cell migration was assessed by the scratch assay,cell cycle distribution and apoptosis were analyzed by flow cytometry,and protein expression levels of tissue inhibitor of metalloproteinases-2(TIMP-2),pro-apoptotic Bcl-2-associated X protein(Bax),cleaved-caspase-3,matrix metalloproteinases(MMP)-2,MMP-9,and anti-apoptotic Bcl-2 were determined by Western blot.A xenograft tumor model was established by subcutaneous injection of H1975 cells into the back of nude mice,which were randomly divided into control,gefitinib(50 mg/kg),and low-,medium-,and high-dose UA(100 mg/kg,200 mg/kg,400 mg/kg)groups.Drugs were administered every three days for 18 consecutive days,and nude mouse body weight and tumor growth were recorded.UPLC-MS/MS was used to determine endogenous metabolite levels in nude mouse serum.Results The IC50 value of UA for inhibiting H1975 cell proliferation at 48 h was 27.8 μmol/L.Compared with the control group,medium-and high-dose UA groups arrested the cell cycle at the G0/G1 phase,and showed increased apoptosis rate and reduced cell migration ability(P＜0.05).Additionally,the protein expression levels of TIMP-2,Bax,and Cleaved Caspase-3 were upregulated,while those of MMP-2,MMP-9,and Bcl-2 were downregulated(P＜0.05).Compared with the low-dose UA group,the high-dose UA group arrested the cell cycle at the G0/G1 phase,and exhibited elevated apoptosis rate,reduced cell migration ability(P＜0.05),and upregulated TIMP-2,Bax,and Cleaved-Caspase-3 protein expression levels,as well as downregulated MMP-2,MMP-9,and Bcl-2 protein expression levels(P＜0.05).In vivo,compared with the control group,all dose groups of UA and the gefitinib group significantly inhibited tumor growth in tumor-xenografted nude mice(P＜0.05).Furthermore,the high-dose UA group demonstrated enhanced tumor growth inhibition compared with the low-and medium-dose UA groups(P＜0.05).Metabolomics analysis identified 22 differential metabolites,including 5S,15S-dihydroxy-6E,8Z,10Z,13E-eicosatetraenoic acid,4-hydroxybenzyl alcohol,and trimethylamine N-oxide,primarily involving pathways related to unsaturated fatty acid biosynthesis,linoleic acid metabolism,and tyrosine metabolism.Conclusion UA exhibits significant inhibitory effects on NSCLC H1975 cells both in vitro and in vivo.Its in vivo inhibitory effects may be associated with the regulation of unsaturated fatty acid biosynthesis,linoleic acid metabolism,and tyrosine metabolism pathways.关键词
熊果酸/H1975细胞/细胞凋亡/细胞增殖/细胞迁移/血清代谢组学Key words
ursolic acid/H1975 cell/apoptosis/cell proliferation/cell migration/serum metabolomics分类
中医学引用本文复制引用
何甜甜,吴柯楠,朱洁,黄成银,侯宝龙,王征,梁艳妮..熊果酸对非小细胞肺癌H1975细胞的体内外抑制作用研究[J].湖南中医药大学学报,2025,45(5):825-835,11.基金项目
陕西省重点产业创新链项目(2018ZDCXL-SF-01-02-02) （2018ZDCXL-SF-01-02-02）
陕西省教育厅创新团队项目(22JP019,24JP047) （22JP019,24JP047）
陕西高校青年创新团队项目(陕教[2019]90号). （陕教[2019]90号）
