文献类型：期刊文献

中文题名：顺铂联合归芪益元膏调节EGFR/MAPK通路对Lewis肺癌荷瘤小鼠的影响

英文题名：Effect of cisplatin combined with Guiqi Yiyuan Ointment on Lewis lung cancer-bearing mice by regulating EGFR/MAPK pathway

作者：张鹏飞[1];王金华[1];梁建庆[1,3];张惠娟[1];李金田[2,3]

第一作者：张鹏飞

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

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2025

卷号：50

期号：2

起止页码：472

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；

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);2022年度中医学一级学科“岐黄英才”导师专项基金硕导项目(ZYXKSD-202211);敦煌医学与转化教育部重点实验室开放课题(DHYX22-06)。

语种：中文

中文关键词：顺铂;归芪益元膏;EGFR/MAPK通路;增殖;周期阻滞

外文关键词：cisplatin;Guiqi Yiyuan Ointment;EGFR/MAPK pathway;proliferation;periodic blockade

摘要：基于表皮生长因子受体(EGFR)/丝裂原活化蛋白激酶(MAPK)通路介导的细胞增殖探讨顺铂联合归芪益元膏对Lewis肺癌荷瘤小鼠的影响。60只雄性C57BL/6小鼠随机选取10只作为空白组,剩余50只为造模组,造模成功后随机分为模型组、顺铂组以及顺铂联合归芪益元膏低、中、高剂量组,每组10只,给药14 d。观察小鼠一般情况,称重、计算脏器指数及抑瘤率;苏木素-伊红(HE)染色观察肿瘤组织病理学形态变化;免疫组化检测瘤组织中Ki-67抗原(Ki-67)和增殖细胞核抗原(PCNA)的阳性率;蛋白免疫印迹法和实时荧光定量聚合酶链式反应(qPCR)检测瘤组织中通路相关蛋白及mRNA表达;流式细胞术检测瘤组织中肿瘤细胞的周期。结果显示,与空白组比较,模型组小鼠一般情况有所恶化;给药14 d后,模型组小鼠体质量、胸腺和脾脏指数均降低。与模型组比较,顺铂组小鼠一般情况恶化,各联用组小鼠有所改善;顺铂组小鼠体质量、胸腺和脾脏指数均降低,各联用组小鼠体质量、胸腺和脾脏指数均升高;各给药组瘤重降低,抑瘤率升高;肿瘤细胞均有不同程度的坏死,且肿瘤细胞紧密度、细胞核和染色质增多情况、核分裂均降低;各给药组Ki-67和PCNA的阳性率,p-EGFR/EGFR、肉瘤病毒癌基因(Ras)、磷酸化Raf-1蛋白激酶(p-Raf-1)/Raf-1、磷酸化丝裂原活化蛋白激酶激酶(p-MEK)/MEK、磷酸化细胞外信号调节激酶(p-ERK)/ERK水平,EGFR、Ras、Raf-1、MEK、ERK mRNA的表达均降低;各给药组肿瘤细胞G0/G1期比例升高,S期比例降低,G2/M期无显著差异。与顺铂组比较,各联用组瘤重降低,抑瘤率升高;肿瘤细胞的坏死及核分裂降低等情况更为显著;Ki-67和PCNA的阳性率,p-EGFR/EGFR、Ras、p-Raf-1/Raf-1、p-MEK/MEK、p-ERK/ERK水平,EGFR、Ras、Raf-1、MEK、ERK mRNA的表达均降低。与顺铂组比较,联用中、高剂量组肿瘤细胞G0/G1期比例升高,S期比例降低;各联用组肿瘤细胞G2/M期比例无显著差异。结果表明顺铂联合归芪益元膏可增强顺铂对荷瘤小鼠的抑瘤作用,其机制可能与抑制EGFR/MAPK通路加速肿瘤细胞G0/G1期阻滞进而抑制肿瘤细胞的增殖有关。同时,研究还表明归芪益元膏可能改善肿瘤对小鼠的损耗以及顺铂化疗带来的毒副作用。
Based on the epidermal growth factor receptor(EGFR)/mitogen-activated protein kinase(MAPK)signaling pathwaymediated cell proliferation,this study explores the effect of cisplatin combined with Guiqi Yiyuan Ointment on Lewis lung cancerbearing mice.A total of 60 male C57BL/6 mice were randomly divided into a blank group with 10 mice and a modeling group with 50 mice.After modeling,they were randomly divided into the model group,cisplatin group,and low-,medium-,and high-dose groups of cisplatin combined with Guiqi Yiyuan Ointment,with 10 mice in each group.After 14 days of medication,the general condition of the mice was observed;body weight was measured,and organ index and tumor inhibition rate were calculated.Hematoxylin-eosin(HE)staining was used to observe the pathological morphology changes in tumor tissue.Immunohistochemistry was used to detect the positive rate of Ki-67 antigen(Ki-67)and proliferating cell nuclear antigen(PCNA)in tumor tissue.Western blot and real time-quantitative polymerase chain reaction(qPCR)were used to detect the expression of related proteins and mRNA in tumor tissue.Flow cytometry was used to detect the cell cycle of tumor cells in tumor tissue.The results showed that compared with that in the blank group,the general condition of mice in the model group deteriorated;the body weight,as well as thymus and spleen index of mice in the model group decreased after 14 days of medication.Compared with that in the model group,the general condition of mice in the cisplatin group deteriorated,while the condition of mice in the combined groups improved;the body weight,as well as thymus and spleen index of mice in the cisplatin group decreased,while the three indicators in the combined groups increased;the tumor weight of each medication group decreased,and the tumor inhibition rate increased;there were varying degrees of necrosis in tumor cells of each medication group,and the tightness of tumor cells,the increase in the number of cell nuclei and chromatin,and mitosis all decreased.The positive rate of Ki-67 and PCNA,as well as the protein expression and ratio of p-EGFR/EGFR,rat sarcoma viral oncogene homolog(Ras),phosphorylated Raf-1 protein kinase(p-Raf-1)/Raf-1,phosphorylated mitogen-activated protein kinase kinase(p-MEK)/MEK,phosphorylated extracellular signal-regulated kinase(p-ERK)/ERK and the mRNA expression of EGFR,Ras,Raf-1,MEK,and ERK all decreased.The proportion of tumor cells in the G0/G1 phase of each medication group increased,and that in the S phase decreased.In addition,there was no significant difference in the G2/M phase.Compared with that of the cisplatin group,the tumor weight of the combined groups decreased,and the tumor inhibition rate increased.The necrosis and mitosis of tumor cells in the combined groups were more pronounced;the positive rate of Ki-67 and PCNA,the protein expression and ratio of p-EGFR/EGFR,Ras,p-Raf-1/Raf-1,p-MEK/MEK,and p-ERK/ERK,as well as the mRNA expression of EGFR,Ras,Raf-1,MEK,and ERK in the combined groups all decreased.The proportion of tumor cells in the G0/G1 phase of the combined medium-and high-dose groups increased,and that in the S phase decreased.There was no significant difference in the proportion of tumor cells of the combined groups in the G2/M phase.This indicates that the combination of cisplatin and Guiqi Yiyuan Ointment can enhance the anti-tumor effect of cisplatin on tumor-bearing mice,and the mechanism may be associated with the inhibition of the EGFR/MAPK pathway,which accelerates the arrest of tumor cells in the G0/G1 phase,thereby inhibiting the proliferation of tumor cells.At the same time,the study also indicates that Guiqi Yiyuan Ointment may reduce the damage of tumors to mice and the toxic side effects brought by cisplatin chemotherapy.