摘要
目的:探讨车前子苷(PMS)对肝细胞癌的治疗效果并对其作用机制进行研究。方法:将人肝细胞癌细胞系HepG2和正常肝细胞系LO-2分别加入160μmol/L的PMS,在37℃,5%CO_(2)的细胞培养箱中培养48 h后,通过BD流式细胞计数仪检测细胞的凋亡情况,通过CCK8实验在0 h、24 h、48 h、72 h检测药物对细胞的毒性情况。通过尾静脉注射HepG2细胞构建小鼠肝细胞癌动物模型,分为HepG2组和HepG2+PMS组,HepG2+PMS组在小鼠发病后每3天灌胃160μmol/LPMS,持续治疗21 d,并通过小鼠活体荧光成像系统(IVIS)记录小鼠肿瘤负荷情况。统计两组小鼠的生存结局,并根据结果绘制Kaplan-Meier曲线。在治疗结束后取两组小鼠肝脏组织切片通过免疫荧光显微镜检测CD206、CD86阳性细胞数,统计M1/M2型巨噬细胞比例。通过体外使用THP1细胞系诱导M1和M2型巨噬细胞,与HepG2细胞孵育后使用免疫荧光检测,并加入PMS观察其对巨噬细胞转化的影响。结果:PMS对正常人肝细胞无细胞毒作用,对人肝癌细胞具有细胞毒作用,可促进肝癌细胞的凋亡,且程度随剂量增加。PMS治疗后的肝细胞癌小鼠生存结局较未治疗组有明显改善(P<0.05)。小鼠肝细胞切片中M2型巨噬细胞的比例极显著降低,M1型巨噬细胞比例显著升高(P<0.05)。THP1诱导后M1型巨噬细胞在PMS的作用下比例较未使用PMS组中升高,M2型巨噬细胞在PMS作用较未使用PMS组中下比例降低(P<0.05)。结论:PMS对肝癌细胞具有细胞毒作用,其主要通过影响肿瘤微环境中的M2型巨噬细胞转化而发挥抗肿瘤效应。
Objective:This study investigates the therapeutic effect of Plantamajoside(PMS)on hepatocellular carcinoma(HCC)and elucidates its underlying mechanism of action.Methods:HepG2 human HCC cells and LO2 normal liver cells were treated with 160μM Plantamajoside and cultured for 48 hours at 37℃in a 5%CO_(2) cell culture incubator.Cell apoptosis was assessed using a BD flow cytometer,and cell toxicity was evaluated at 0 h、24 h、48 h and 72 h using the CCK8 assay.An HCC mouse model was established by tail vein injection of HepG2 cells,divided into the HepG2 group and HepG2+PMS group.The HepG2+PMS group received 160μM Plantamajoside via oral gavage every three days after disease onset,with treatment lasting for 21 days.Tum or burden in mice was monitored using the In Vivo Imaging System(IVIS).Survival outcomes were analyzed,and Kaplan-Meier curves were generated.Post-treatment liver tissues from the two groups of mice were collected,and immunofluorescence microscopy was used to quantify CD206 and CD86-positive cells and determine the M1/M2 macrophage ratio.THP1 cells were induced to differentiate into M1 and M2 macrophages,cocultured with HepG2 cells,and subjected to immunofluorescence staining to assess the effect of Plantamajoside on macrophage polarization.Results:Plantamajoside exhibited no cytotoxic effects on normal liver cells but demonstrated cytotoxicity against HepG2 cells,inducing apoptosis in a dose-dependent manner.Treatment with Plantamajoside significantly improved survival outcomes in HCC mice compared to the untreated group(P<0.05).Immunofluorescence analysis of liver tissue revealed a marked decrease in M2 macrophage proportion and a significant increase in M1 macrophage proportion post-treatment(P<0.05).Plantamajoside treatment led to an elevated proportion of M1 macrophages and a decreased proportion of M2 macrophages compared to the untreated group in the THP1 coculture system(P<0.05).Conclusion:Plantamajoside exerts cytotoxic effects on HCC cells,primarily by influencing the transformation of M2 macrophages within the tumor microenvironment to exert antitum or effects.
作者
李畅
陈晋平
岳阳
李金岩
郑俊华
焦莹
LI Chang;CHEN Jin-Ping;YUE-yang(Department of Radiology,Langfang Hospital of Traditional Chinese medicine(Hebei Langfang,065000),China;不详)
出处
《中西医结合肝病杂志》
2025年第11期1369-1373,共5页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
河北省中医药管理局中医药类科研计划(No.2022298)。
关键词
车前子苷
肝细胞癌
肿瘤微环境
巨噬细胞
plantamajoside
hepatocellular carcinoma
tumor microenvironment
macrophages
