世界科学技术-中医药现代化2026，Vol.28Issue(5)：1503-1516,14.DOI:10.11842/wst.20250328003
基于NF-κB信号通路对M1型巨噬细胞极化的影响探讨柴芪益肝颗粒调控对HepG2细胞的作用机制
Explore the Mechanism of Chaiqi Yigan Granule Regulation on HepG2 Cells Based on the Effect of NF-κB Signaling Pathway on the Polarization of M1-type Macrophages
摘要
Abstract
Objective To investigate the mechanism of ChaiQiYiGanKeLi(CQYGKL)modulating NF-κB signaling pathway mediating M1-type macrophage polarization on HepG2 cells.Methods Tissues from 20 patients with hepatocellular carcinoma(including hepatocellular carcinoma tissues and their paracancerous tissues)were collected,and the expression of M1-type macrophage-associated markers(CD16,CD68)in hepatocellular carcinoma tissues and their paracancerous tissues were detected by immunohistochemistry.Preparation of CQYGKL-containing serum was prepared and set aside.The survival rate of HepG2 cells after intervention with different concentrations of CQYGKL-containing serum was detected by CCK8 method.A cell co-culture system was established,and the effect of CQYGKL on the migration of HepG2 cells in the co-culture system was detected by cell scratch assay.The cells in the co-culture system were intervened with different factors and were divided into:control group,blank serum group,CQYGKL group,BAY 11-7082 group,PMA group and PMA+CQYGKL group,and the cells in the lower chamber of Transwell were collected after 48 h of intervention for the subsequent experiments.①RT-qPCR was used to detect the expression of Caspase-3,Bcl-2,N-cadherin,E-cadherin,Vimentin,MMP7,CD68,CD16,iNOS and NF-κBp65 mRNA in the cells of each group.②The expression of Caspase-3,Bcl-2,N-cadherin,E-cadherin,Vimentin,MMP7,CD68,CD16,P-NF-κBp65 and NF-κBp65 proteins were detected by Western blot in each group of cells.③The expression of iNOS in the cell supernatant of each group was detected by ELISA assay.Results The expression of M1-type macrophage markers(CD16 and CD68)was significantly lower in hepatocellular carcinoma tissues compared with that in paraneoplastic tissues(P<0.01).The results of CCK-8 experiment showed that after 24 h of intervention,the survival rate of HepG2 cells was significantly decreased when the concentration of CQYGKL-containing serum was more than 20%compared with the control group(P<0.01);And after 48 h of intervention,the survival rate of HepG2 cells was significantly decreased when the concentration of CQYGKL-containing serum was more than 10%compared with the control group(P<0.01).The results of the scratch experiment showed that compared with the control group,there was no significant decrease in the healing rate of the blank serum group for either 24 h or 48 h of intervention(P>0.05),whereas CQYGKL significantly reduced the healing rate of HepG2 cells for both 24 and 48 h of intervention(P<0.01).Following intervention with different factors in a co-culture system,no statistically significant differences in mRNA and protein expression levels of Caspase-3,Bcl-2,N-cadherin,E-cadherin,Vimentin,MMP7,CD68,CD16,iNOS,and P-NF-κBp65 were observed between the control group and the blank serum group,nor between the CQYGKL group and the BAY 11-7082 group(P>0.05).Compared with the control group,both CQYGKL and BAY 11-7082 significantly downregulated the mRNA and protein expression of Bcl-2,N-cadherin,Vimentin,MMP7,and P-NF-κBp65(P<0.01 or P<0.05),while upregulating the expression of Caspase-3,E-cadherin,CD68,CD16,and iNOS(P<0.01).In contrast,PMA promoted the expression of Bcl-2,N-cadherin,Vimentin,MMP7,and P-NF-κBp65(P<0.01 or P<0.05)and suppressed the expression of Caspase-3,E-cadherin,CD68,CD16,and iNOS(P<0.01 or P<0.05).Notably,compared with the PMA group,co-treatment with CQYGKL and PMA inhibited the expression of Bcl-2,N-cadherin,Vimentin,MMP7,and P-NF-κBp65(P<0.01 or P<0.05),and enhanced the expression of Caspase-3,E-cadherin,CD68,CD16,and iNOS(P<0.01 or P<0.05).Conclusion CQYGKL can promote the apoptosis of HepG2 cells in the co-culture system and inhibit their metastasis.The mechanism of action may be related to its ability to inhibit the activation of the NF-κB signaling pathway in the co-culture system and further promote the polarization of M0 macrophages to M1-type macrophages in the co-culture system.关键词
柴芪益肝颗粒/HepG2细胞/NF-κB信号通路/巨噬细胞极化/凋亡/转移Key words
ChaiQiYiGanKeLi/HepG2 cells/NF-κB signaling pathway/Macrophage polarization/Apoptosis/Metastasis分类
医药卫生引用本文复制引用
李峰,马贵萍,苏晓鹏,刘博文,李晓斌,吴芬芳,冉云,胡世平..基于NF-κB信号通路对M1型巨噬细胞极化的影响探讨柴芪益肝颗粒调控对HepG2细胞的作用机制[J].世界科学技术-中医药现代化,2026,28(5):1503-1516,14.基金项目
广东省科学技术厅广东省自然科学基金面上项目(2025A1515011796):基于mTORC1介导的肿瘤细胞铁死亡调控肿瘤相关巨噬细胞极化探讨柴芪益肝颗粒治疗肝细胞癌的作用机制,负责人:胡世平 （2025A1515011796）
深圳市科技计划面上项目(JCYJ20220530172812028):基于NF-KB信号通路对肿瘤微环境中M1-M2型巨噬细胞极化的影响探讨柴芪益肝颗粒防止乙肝肝癌转移的作用机制,负责人:胡世平 （JCYJ20220530172812028）
北京中医药大学研究生自主科研课题项目(ZJKT2023041):基于NF-KB信号通路对巨噬细胞极化的影响探讨柴芪益肝颗粒对肝癌的作用机制,负责人:李峰. （ZJKT2023041）
