摘要
目的探究健脾益气方对脂多糖(LPS)联合三磷酸腺苷(ATP)刺激下的构造的炎症微环境下人肝癌Huh-7细胞的增殖、侵袭及凋亡作用影响,并检测细胞中白细胞介素(IL)-1β、IL^(-1)8的表达水平。方法构建健脾益气方含药血清,采用向细胞培养基中加入LPS与ATP共培养的方法刺激肝癌细胞并构建炎症微环境。将细胞分为空白组、模型组、VX-765组(10μmol·L^(-1))、低浓度中药组(15%健脾益气方低剂量含药血清)、中浓度中药组(15%健脾益气方中剂量含药血清)、高浓度中药组(15%健脾益气方高浓度含药血清)。细胞干预后采用CCK-8法检测各组细胞的增殖水平;通过Annexin V-FITC/PI法检测各组细胞的凋亡率;采用Transwell法检测各组细胞侵袭水平;采用PI单染色法检测各组细胞的周期水平;采用ELISA法、Western blot法检测Huh-7细胞中IL^(-1)b及1L^(-1)8表达水平。结果与空白组相比,采用LPS与ATP刺激后的模型组Huh-7细胞的增殖水平、侵袭水平与IL^(-1)β及1L^(-1)8表达水平更高(P<0.05),凋亡水平更低(P<0.05)。与其他组别相比,中浓度中药组与高浓度中药组能有效抑制Huh-7细胞的增殖水平与侵袭水平,阻滞细胞增殖周期,降低细胞存活率(P<0.05),并能显著诱导Huh-7细胞凋亡(P<0.05),并且降低Huh-7细胞中IL^(-1)β及1L^(-1)8表达水平(P<0.05)。结论中浓度及高浓度的健脾益气方含药血清能够通过改善肝癌Huh-7细胞中的炎症微环境实现对人肝癌Huh-7细胞的抑制增殖与侵袭,阻滞细胞周期和诱导凋亡作用,其作用机制可能与抑制肝癌细胞分析炎症因子IL^(-1)β及1L^(-1)8水平以实现对原发性肝癌的治疗目的。
Objective To explore the effect of JianpiYiqi Decoction on the proliferation,invasion and apoptosis of Human hepatoma Huh-7 cells under the constructed inflammatory microenvironment stimulated by lipopolysaccharide(LPS)and adenosine triphosphate(ATP),and to detect the interleukin-1β(IL^(-1)β)and interleukin-18(IL^(-1)8)Expression level in cells.Methods Construction drug-containing serum of JianpiYiqi.Adopt LPS and ATP were added to the cell culture medium to stimulate hepatoma cells and construct the inflammatory microenvironment.The cells were divided into blank group,model group,VX-765 group(10μmol·L^(-1)),low concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction low dosedrug-containing serum),Medium concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction Medium dose drug-containing serum),and high concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction high dose drug-containing serum).Adopt CCK-8 method was used to detect the proliferation level of cells in each group after cell intervention;The apoptosis rate of cells in each group was detected by Annexin V-FITC/PI method;Adopt Transwell method was used to detect the level of cell invasion in each group;adopt PI single staining was used to detect the cell cycle level in each group;Adopt ELISA method and Western blot method were used to detection of IL^(-1)βand 1l-18 expression level in Huh-7 cells.Results Compared with the blank group,in the model group stimulated by LPS and ATP,the proliferation level,invasion level,IL^(-1)βand IL^(-1)8 expression level of Huh-7 cells were higher(P<0.05),and the apoptosis level was lower(P<0.05).Compared with other groups,the Medium concentration of traditional Chinese medicine group and the high concentration of traditional Chinese medicine group could effectively inhibit the proliferation and invasion level of Huh-7 cells,block the cell proliferation cycle,reduce the cell survival rate(P<0.05),significantly induce apoptosis of Huh-7 cells(P<0.05),and reduce the expression levels of IL^(-1)βand IL^(-1)8 in Huh-7 cells(P<0.05).Conclusion The drug-containing serum JianpiYiqi.The medium and high concentrations of JianpiYiqi Decoction drug-containing serum can inhibit the proliferation and invasion,block the cell cycle and induce apoptosis of human liver cancer Huh-7 cells by improving the inflammatory microenvironment in liver cancer Huh-7 cells.Its mechanism may be related to the inhibition of liver cancer cells.Analyze the levels of inflammatory factors IL^(-1)βand IL^(-1)8 to achieve the therapeutic purpose of primary liver cancer.
作者
陈琳
蒋锐沅
欧少甫
梁天坚
罗淑娟
刘磊
张萌
蒋筱
Chen Lin;Jiang Ruiyuan;Ou Shaofu;Liang Tianjian;Luo Shujuan;Liu Lei;Zhang Meng;Jiang Xiao(Graduate School of Guangxi University of Chinese Medicine,Nanning 530000,China;Cancer Hospital Affiliated to Chinese Academy of Sciences,Hangzhou 310000,China;Second Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou 310000,China;Sains New Medical College of Guangxi University of Traditional Chinese Medicine,Nanning 530222,China;School of Basic Medicine,Guangxi University of Chinese Medicine,Nanning 530000,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第11期3704-3714,共11页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
广西高校中青年教师科研基础能力提升项目(2022CX001):基于“脾虚”理论探讨健脾益气法促进消化道肿瘤细胞焦亡的机制研究,负责人:梁天坚
广西中医药大学博士科研启动基金项目(2020BS010):《伤寒杂病论》“药-方-证-机”理论指导下健脾益气方干预肝癌的分子机制预测与验证,负责人:张萌
广西自然科学基金项目面上项目(2018GXNSFAA281045):Caspase-1/IRAKs/NF-kB信号通路在肝癌细胞中的作用及健脾治法干预研究,负责人:蒋筱
关键词
肝癌Huh-7细胞
炎症微环境
脾气亏虚
健脾益气方
Liver cancer Huh-7 cells
Inflammatory microenvironment
Deficiency of temper
JianpiYiqi decoction
作者简介
通讯作者:蒋筱,硕士,教授,硕士研究生导师,广西名中医,主要研究方向:中医治则治法理论与应用研究;通讯作者:张萌,博士,讲师,主要研究方向:中医治则治法理论与应用研究。
