摘要
目的探讨丹皮酚调控核因子κB(NF-κB)/缺氧诱导因子1α(HIF-1α)信号通路介导的有氧糖酵解从而抑制膀胱癌T24细胞迁移的作用及机制。方法将T24细胞分为对照组、顺铂组(阳性对照,3.001μg/mL)和丹皮酚低、中、高剂量组(100、200、400μg/mL),给药干预24 h后,检测丹皮酚对T24细胞迁移能力的影响(以细胞划痕愈合率表示),检测丹皮酚对T24细胞线粒体膜电位的影响(以红绿荧光强度比值表示),检测T24细胞中腺苷三磷酸(ATP)浓度和乳酸含量,检测细胞中NF-κB/HIF-1α信号通路、迁移相关蛋白以及有氧糖酵解关键酶的表达水平。结果与对照组比较,丹皮酚中、高剂量组和顺铂组细胞划痕愈合率均显著降低(P<0.01);丹皮酚各剂量组细胞中NF-κB/HIF-1α信号通路相关蛋白NF-κB、HIF-1α,迁移相关蛋白基质金属蛋白酶2(MMP2)、MMP9、血管内皮生长因子,有氧糖酵解关键酶葡萄糖转运蛋白1、己糖激酶2、丙酮酸激酶M2表达水平均有不同程度降低,大部分差异有统计学意义(P<0.05或P<0.01);丹皮酚中、高剂量组细胞线粒体红绿荧光强度比值均显著降低(P<0.01);丹皮酚高剂量组细胞ATP浓度以及丹皮酚各剂量组细胞乳酸含量均显著降低(P<0.05或P<0.01)。结论丹皮酚可抑制膀胱癌T24细胞的迁移,其作用机制可能与抑制NF-κB/HIF-1α信号通路、下调有氧糖酵解关键酶活性有关。
OBJECTIVE To investigate the role and mechanism of paeonol in inhibiting the migration of bladder cancer T24 cells by regulating nuclear factorκB(NF-κB)/hypoxia-inducible factor-1α(HIF-1α)-mediated aerobic glycolysis.METHODS T24 cells were divided into control group,cisplatin group(positive control,3.001μg/mL),and paeonol low-,medium-and high-dose groups(100,200,400μg/mL),respectively.After 24 h of administration intervention,the effect of paeonol on the migration ability of T24 cells was detected(expressed by the cell scratch wound healing rate).The effect of paeonol on the mitochondrial membrane potential of T24 cells was detected(expressed by the ratio of red/green fluorescence intensity).Cellular adenosine triphosphate(ATP)levels and lactate content in T24 cells were measured.The levels of NF-κB/HIF-1αsignaling pathway,the expression of migration-related proteins,and key enzymes involved in aerobic glycolysis in the cells were all determined.RESULTS Compared with the control group,the cell scratch wound healing rates in the paeonol medium-and high-dose groups and the cisplatin group were decreased significantly(P<0.01);in the paeonol groups,the expression levels of NF-κB/HIF-1αsignaling pathway-related proteins such as NF-κB and HIF-1α,migration-related proteins such as matrix metalloproteinase 2(MMP2),MMP9,and vascular endothelial growth factor,as well as key enzymes involved in aerobic glycolysis such as glucose transporter 1,hexokinase 2 and pyruvate kinase isozyme type M2,were all reduced to varying degrees in the cells,most of these reductions showed statistically significant differences(P<0.05 or P<0.01);the ratio of red/green fluorescence intensity in mitochondria of cells in the medium-and high-dose paeonol groups were significantly decreased(P<0.01);the ATP concentration in cells of the paeonol high-dose group,and the lactate content in cells across all paeonol groups were significantly decreased(P<0.05 or P<0.01).CONCLUSIONS Paeonol significantly inhibits the migration of bladder cancer T24 cells,and its mechanism of action may be related to the inhibition of the NF-κB/HIF-1αsignaling pathway,and the down-regulation of key enzyme activities involved in aerobic glycolysis.
作者
艾欣瑶
陈雯佳
陈曦
王颖峥
王英豪
黄美霞
AI Xinyao;CHEN Wenjia;CHEN Xi;WANG Yingzheng;WANG Yinghao;HUANG Meixia(School of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)
出处
《中国药房》
北大核心
2025年第15期1871-1875,共5页
China Pharmacy
基金
国家中医药管理局临床中药学高水平中医药重点学科建设项目(No.zyyzdxk-2023107)
福建省自然科学基金项目(No.2023J01857)
福建中医药大学基础类学科科研提升计划(No.XJC2022013)
福建中医药大学省级财政资助专项(No.X2023005-财政专项)。
作者简介
第一作者:艾欣瑶,硕士研究生。研究方向:中药药理与临床。E-mail:1791876695@qq.com;通信作者:黄美霞,副教授,硕士生导师。研究方向:中药药理与临床。E-mail:2008009@fjtcm.edu.cn。
