归芪益元膏联合顺铂通过内质网应激途径和线粒体凋亡途径对Lewis肺癌小鼠的影响  

Effect of Guiqi Yiyuan Ointment Combined with Cisplatin on Mice with Lewis Lung Cancer Through Endoplasmic Reticulum Stress Pathway and Mitochondrial Apoptosis Pathway

作　　者：孔思琪 李金田 李娟[3] 梁建庆 张毅 张悦 袁超 KONG Siqi;LI Jintian;LI Juan;LIANG Jianqing;ZHANG Yi;ZHANG Yue;YUAN Chao(The Clinical College of Traditional Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730101,China;The Key Laboratory of the Ministry of Education of Dunhuang Medicine and Transformation,Lanzhou 730000,China;School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730101,China)

机构地区：[1]甘肃中医药大学中医临床学院,兰州730101 [2]敦煌医学与转化教育部重点实验室,兰州730000 [3]甘肃中医药大学基础医学院,兰州730101

出　　处：《中国实验方剂学杂志》2024年第20期54-61,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82160872);甘肃省教育厅“双一流”科研重点项目(GSSYLXM-05);甘肃省中医药研究中心开放课题(zyzx-2020-zx17);2022年度中医学一级学科“岐黄英才”导师专项基金博导项目(ZYXKBD-202202)。

摘　　要：目的:探讨归芪益元膏联合顺铂通过内质网应激途径和线粒体凋亡途径对Lewis肺癌小鼠的影响。方法:将50只SPF级雄性C57BL/6小鼠,随机分为模型组、顺铂组(0.005 g·kg^(-1))、归芪益元膏联合顺铂(联合)低、中、高剂量组(0.005+1.6 g·kg^(-1)、0.005+3.3 g·kg^(-1)、0.005+6.6 g·kg^(-1))。各组小鼠腋下接种Lewis细胞悬液,构建Lewis肺癌移植瘤小鼠模型。连续给药14 d后取材,称量小鼠体质量,剥离瘤体称量瘤体质量,计算脏器指数和抑瘤率;苏木素-伊红(HE)染色观察肿瘤组织病理形态变化;流式细胞仪检测肿瘤细胞凋亡率和活性氧(ROS)比率;蛋白免疫印迹法(Western blot)检测内质网应激途径中葡萄糖调节蛋白78(GRP78)、磷酸化蛋白激酶R样内质网激酶(p-PERK)、活化转录因子4(ATF4)、凋亡蛋白C/EBP同源蛋白(CHOP)和线粒体凋亡途径中胱天蛋白酶-9(Caspase-9)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达情况。结果:与模型组比较,归芪益元膏联合顺铂组小鼠毛色较光泽,精神反应状态较好;各给药组肿瘤质量均降低(P<0.05),抑瘤率均升高(P<0.05);联合组胸腺、脾脏指数均升高(P<0.05);各给药组肿瘤组织病理形态改变明显,异型性逐渐降低,联合高剂量组肿瘤组织被大量破坏;各给药组肿瘤细胞凋亡率及ROS生成率均升高(P<0.05);各给药组肿瘤组织中Bcl-2蛋白表达水平逐渐降低(P<0.05),而GRP78、p-PERK、ATF4、CHOP、Bax、Caspase-9蛋白表达水平明显升高(P<0.05)。与顺铂组比较,联合组肿瘤质量均降低(P<0.05),联合中、高剂量组抑瘤率均升高(P<0.05);联合组胸腺指数和脾脏指数、肿瘤细胞凋亡率、ROS比率明显升高(P<0.05);联合中、高剂量组GRP78、p-PERK、Bax、Caspase-9蛋白表达水平明显升高(P<0.05);联合组ATF4、CHOP蛋白表达水平升高(P<0.05),Bcl-2蛋白表达水平逐渐降低(P<0.05)。结论:归芪益元膏联合顺铂可对Lewis肺癌小鼠产生抑瘤作用,Objective:To explore the effects of Guiqi Yiyuan ointment combined with cisplatin on mice with Lewis lung cancer through the endoplasmic reticulum stress pathway and mitochondrial apoptosis pathway.Method:Fifty SPF male C57BL/6 mice were randomly divided into the model group,cisplatin group(0.005 g·kg^(-1)),and low,medium,and high dose groups of Guiqi Yiyuan ointment combined with cisplatin(0.005+1.6 g·kg^(-1),0.005+3.3 g·kg^(-1),and 0.005+6.6 g·kg^(-1)).Lewis cell suspension was inoculated under the axilla of mice in each group to construct the Lewis lung cancer xenograft mouse model.After continuous administration for 14 days,the mice were sacrificed.The body weight of the mice was measured,and the tumor weight was measured after the tumors were removed.The organ index and tumor inhibition rate were calculated.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in tumor tissue.Flow cytometry was used to detect the apoptosis rate of tumor cells and the ratio of reactive oxygen species(ROS).Western blot was used to detect the expression of glucose-regulated protein 78(GRP78),phosphorylated activated protein kinase R-like endoplasmic reticulum kinase(p-PERK),activated transcription factor 4(ATF4),and apoptosis protein C/EBP homologous protein(CHOP)in the endoplasmic reticulum stress pathway,as well as cysteine aspartate protease-9(Caspase-9),B-cell lymphoma-2(Bcl-2),and Bcl-2 associated X protein(Bax)in the mitochondrial apoptosis pathway.Result:Compared with those in the model group,the mice in the groups of Guiqi Yiyuan Ointment combined with cisplatin had shinier fur and better mental response status.Tumor mass was reduced in all treatment groups(P<0.05),and tumor inhibition rate was increased in all treatment groups(P<0.05).The thymus and spleen indices of the combined group were increased(P<0.05),and obvious pathological changes were observed in the tumor tissue of all treatment groups,with a gradual decrease in heteromorphism.Destruction of massive tumor tissue was observed in the

关 键 词：归芪益元膏 肺癌 顺铂 线粒体凋亡途径 内质网应激途径 

分 类 号：R2-0[医药卫生—中医学] R22R242R285.5R734.2