2025 年第 4 期 第 20 卷

夏枯草醇提取物木犀草素抑制磷脂酰肌醇-3-激酶/蛋白激酶B通路进而抑制视神经胶质瘤细胞增殖和迁移的观察研究

Observational study on the inhibitory effect of luteolin, an alcohol extract from Prunella vulgaris, on the phosphatidylinositol 3-kinase/protein kinase B pathway, subsequently suppressing the proliferation and migration of optic glioma cells

作者：杨宝义1 叶向梅2 吴限1 孙河1

英文作者：Yang Baoyi1 Ye Xiangmei2 Wu Xian1 Sun He1

单位：1黑龙江中医药大学附属第一医院外三科，哈尔滨150040；2哈尔滨医科大学附属第一医院血液肿瘤中心，哈尔滨150001

英文单位：1Third Department of Surgery First Affiliated Hospital Heilongjiang University of Chinese Medicine Harbin 150040 China; 2Blood Tumor Center the First Affiliated Hospital Harbin Medical University Harbin 150001 China

关键词：视神经胶质瘤；木犀草素；细胞增殖；细胞迁移；蛋白表达

英文关键词：Opticnerveglioma;Luteolin;Cellproliferation;Cellmigration;Proteinexpression

• 摘要：

• 目的 探讨夏枯草醇提取物中活性成分木犀草素对视神经胶质瘤（ONG）细胞增殖和迁移能力的影响，以揭示其潜在治疗肿瘤的作用机制。方法 培养 U87-MG 胶质瘤细胞，进行分组处理。对照组给予二甲亚砜处理，10、20、40、60 μmol/L木犀草素处理组分别给予10、20、40及60 μmol/L木犀草素处理，替莫唑胺处理组给予100 μmol/L替莫唑胺处理。利用划痕试验、克隆形成实验和细胞计数盒8实验检测不同处理组 U87-MG 细胞的增殖、迁移和克隆形成能力；利用实时荧光定量聚合酶链反应、蛋白质印迹法检测磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B(AKT)通路相关基因PI3K催化亚基γ（PIK3CG）、AKT、波形蛋白、N-钙黏蛋白的mRNA及蛋白表达。结果 木犀草素能显著抑制U87-MG胶质瘤细胞的迁移、克隆形成及细胞的增殖能力，与对照组比较差异均有统计学意义（均P<0.05），并且随着木犀草素浓度的增加抑制性逐渐增强；替莫唑胺组抑制作用明显低于木犀草素组（均P<0.05）。10、20、40、60 μmol/L木犀草素处理组及替莫唑胺处理组处理24 h后胶质瘤U87-MG细胞PI3K/AKT通路基因PIK3CG mRNA表达明显低于对照组［（0.236±0.016）、（0.199±0.016）、（0.143±0.058）、（0.103±0.014）、（0.458±0.014）比（0.977±0.023）］（均P<0.05），随着木犀草素浓度的增加PIK3CG mRNA表达逐渐降低，替莫唑胺组虽然没有木犀草素组降低幅度大，但与对照组的差异也超过2倍；而AKT、波形蛋白、N-钙黏蛋白 mRNA表达受影响不大。 蛋白质印迹实验结果显示与对照组相比，各浓度木犀草素组PIK3CG蛋白表达水平也发生了显著降低，且木犀草素组随着浓度的增加PIK3CG蛋白表达逐渐降低，替莫唑胺组虽然也降低，但降低幅度没有木犀草素组明显，此蛋白检测结果与mRNA结果一致。结论 本研究证实了木犀草素能够有效抑制胶质瘤细胞的增殖和迁移，且可能通过PIK3CG低表达调控PI3K/AKT通路，进而对ONG细胞增殖和迁移产生影响。这一研究结果为夏枯草醇提取物木犀草素治疗ONG提供了实验依据，揭示了作为治疗ONG潜在药物的木犀草素的应用价值，为其临床转化提供了较为扎实的理论基础。

• Objective To investigate the effect of luteolin, the active ingredient in the alcohol extract of Prunella vulgaris, on the proliferation and migration of optic nerve glioma (ONG) cells, and to reveal its potential mechanism of action for tumor treatment. Methods The cultured U87-MG glioma cells were grouped for treatment. The control group was treated with dimethyl sulfoxide, and the 10, 20, 40 and 60 μmol/L luteolin treatment groups were treated with 10, 20, 40 and 60 μmol/L luteolin, respectively. The temozolomide treatment group was treated with 100 μmol/L temozolomide. The proliferation, migration and colony formation ability of U87-MG cells in different treatment groups were detected by scratch assay, colony formation assay and cell counting kit-8 assay. Real-time fluorescent quantitative polymerase chain reaction and Western blotting were used to detect the mRNA and protein expression of phosphatidylinositol-3-kinase (PI3K)/protein kinase B(AKT) pathway-related genes, PI3K catalytic subunit γ (PIK3CG), AKT, vimentin and N-cadherin. Results Luteolin could significantly inhibit the migration, clonogenesis and proliferation of U87-MG glioma cells, and the differences were statistically significant compared with the control group (all P<0.05). With the increase of luteolin concentration, the inhibitory effect was gradually enhanced, and the inhibitory effect of temozolomide group was significantly lower than that of luteolin group (all P<0.05). The PI3K/AKT pathway gene PIK3CG mRNA expressions in glioma U87-MG cells treated with 10, 20, 40, 60 μmol/L luteolin and temozolomide for 24 h were significantly lower than those in the control group ［(0.236±0.016), (0.199±0.016), (0.143±0.058), (0.103±0.014), (0.458±0.014) vs (0.977±0.023)］(all P<0.05). With the increase of luteolin concentration, the expression of PIK3CG mRNA decreased gradually. Although the decrease of temozolomide group was not as great as that of luteolin group, the difference between temozolomide group and control group was more than 2 times. However, the expressions of AKT, vimentin and N-cadherin mRNA were not affected. Western blot results  showed that compared with the control group, the expression level of PIK3CG protein in luteolin groups was also significantly decreased. The expression of PIK3CG protein in luteolin group was gradually decreased with the increase of luteolin concentration, and the expression of PIK3CG protein in temozolomide group was also decreased, but the reduction ratio was not as obvious as that in luteolin group. The results of this protein detection were consistent with the results of mRNA. Conclusion This study confirmed that luteolin can effectively inhibit the proliferation and migration of glioma cells, and may regulate the PI3K/AKT pathway through the low expression of PIK3CG, thereby affecting the proliferation and migration of ONG cells. This study provides an experimental basis for the treatment of ONG with luteolin from the alcohol extract of Prunella vulgaris, reveals the application value of luteolin as a potential drug for the treatment of ONG, and provides a solid theoretical basis for its clinical transformation.