地榆槐花汤通过调控p38 MAPK/p53通路对人结直肠癌HCT116细胞增殖和凋亡的影响  

Effects of Diyu Huaihua decoction on the proliferation and apoptosis of Human Colorectal Cancer HCT116 Cells by Regulating the p38 MAPK/p53 Pathway

作　　者：于春雪 陈楠 丛珊 禹晶[1] 王静 毛颖 梁雪琦 YU Chunxue;CHEN Nan;CONG Shan;YU Jing;WANG Jing;MAO Ying;LIANG Xueqi(The Second Affiliated Hospital of Qiqihar Medical College,Qiqihar 161006,China;Traditional Chinese Medicine Room of Daqing Drug Inspection and Testing Center,Daqing 163311,China;Institute of Traditional Chinese Medicine,Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150001,China)

机构地区：[1]齐齐哈尔医学院附属第二医院,齐齐哈尔161006 [2]大庆市药品检验检测中心中药室,大庆163311 [3]黑龙江省中医药科学院中药所,哈尔滨150001

出　　处：《世界科学技术-中医药现代化》2024年第2期294-301,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(2021-KYYWF-0370):地榆槐花汤对人结肠癌HCT116细胞的影响;负责人:于春雪;黑龙江省中医药科研项目(ZHY2020-045):复合纳豆山楂固体饮料对小儿厌食症动物模型大鼠的影响;负责人:王静;黑龙江省卫生健康委科研项目(20221313010628):MiRNA和lncRNA遗传多态性与阿司匹林治疗后脑梗死预后的关系;负责人:丛珊。

摘　　要：目的探讨地榆槐花汤对人结直肠癌HCT116细胞增殖和凋亡的影响及机制。方法将HCT116细胞分为空白对照组(空白血清)、阳性对照组(奥沙利铂含药血清)、地榆槐花汤低、中、高剂量组(10%、15%、20%地榆槐花汤含药血清),给予相应含药血清干预,MTT检测细胞增殖活性;平板克隆形成实验检测各组细胞克隆形成能力;Annexin V-FITC/PI法检测细胞的凋亡水平;Hoechst 33258染色观察细胞形态变化;Western blot检测细胞中Bax、Bcl-2、Cleaved caspase-3、p38MAPK、p-p38MAPK(Thr180/Tyr182)、MDM2和p53等蛋白表达水平。进一步将HCT116细胞分为空白对照组(空白血清)、地榆槐花汤组(20%地榆槐花汤含药血清)、p38MAPK抑制剂TAK-715组(10 mmol·L^(-1) TAK-715)和联合组(20%地榆槐花汤含药血清+10 mmol·L^(-1) TAK-715),采用20%地榆槐花汤含药血清和10 mmol·L^(-1) TAK-715进行干预,MTT检测各组细胞增殖活性;AnnexinV-FITC/PI法检测细胞凋亡水平;Westernblot检测细胞中p38MAPK、pp38MAPK、MDM2、p53等蛋白表达水平。结果与空白对照组相比,地榆槐花汤中、高剂量组及阳性对照组HCT116细胞的增殖活性和克隆形成能力显著降低(P<0.01),细胞核呈皱缩、碎块状变化,细胞凋亡水平显著增加(P<0.01),Bax/Bcl-2、Cleaved caspase-3、p-p38MAPK/p38MAPK和p53等蛋白表达水平显著上调(P<0.05),MDM2蛋白表达水平显著下调(P<0.01)。然而,联合TAK-715处理可明显降低地榆槐花汤对HCT116细胞增殖的抑制作用和凋亡的诱导作用(P<0.05)。结论地榆槐花汤可通过调控p38 MAPK/p53通路抑制HCT116细胞的增殖,并诱导细胞凋亡。Objective To explore the effects and mechanism of Diyu Huaihua decoction on the proliferation and apoptosis of human colorectal cancer HCT116 cells.Methods HCT116 cells were divided into blank control group(blank serum),positive control group(oxaliplatin-containing serum),Diyu huaihua decoction low-dose,medium-dose and high-dose groups(10%,15%,20%Diyu Huaihua decoction-containing serum).The corresponding drug-containing serum were given for intervention,the cell proliferation activity was detected by MTT.The clone formation ability of each group of cells was detected by the monoclonal formation assay Annexin V-FITC/PI method was used to detecte the level of apoptosis;Hoechst 33258 staining was used to observe cell morphological changes;Western blot was used to detect the protein expression levels of Bax,Bcl-2,cleaved caspase-3,p38MAPK,p-p38MAPK(Thr180/Tyr182),MDM2 and p53 in cells.HCT116 cells were further divided into blank control group(blank serum),Diyu Huaihua decoction group(20%Diyu Huaihua decoction containing serum),p38MAPK inhibitor TAK-715 group(10 mmol·L^(-1) TAK-715)and combination group(20%Diyu Huaihua decoction containing serum+10 mmol·L^(-1) TAK-715).The 20%Diyu Huaihua decoction-containing serum and 10 mmol·L^(-1) TAK-715 were used for intervention.The proliferation activity of cells in each group was detected by MTT;the apoptosis level was detected by Annexin V-FITC/PI method;the protein expression levels of p38MAPK,p-p38MAPK,MDM2 and p53 in the cells were detected by Western blot.Results Compared with the blank control group,the proliferation activity and colony formation ability of HCT116 cells in the Diyuhuaihua decoction medium and high dose groups and the positive control group were significantly decreased(P<0.01).The nucleus showed the changes of shrinking and fragment,the level of apoptosis was significantly increased(P<0.01),and the protein expression levels of Bax/Bcl-2,cleaved caspase-3,p-p38MAPK/p38MAPK and p53 were significantly increased(P<0.05).The protein expression levels of MDM

关 键 词：地榆槐花汤 结直肠癌 细胞增殖 细胞凋亡 p38 MAPK/p53通路 

分 类 号：R285.5[医药卫生—中药学]