摘要
目的探究六君子汤对食管癌模型小鼠治疗作用机制。方法采用4-硝基喹啉-1-氧化物(4-nitroquinolin-1-oxide,4NQO)诱导法构建食管癌小鼠模型。实验分为空白组、模型组、六君子汤低浓度组、六君子汤中浓度组和六君子汤高浓度组。苏木素伊红(Hematoxylin-eosin,HE)染色观察小鼠食管病理改变;免疫组化检测小鼠食管组织中α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、波形蛋白(Vimentin)蛋白表达;实时定量基因扩增荧光检测系统(real-time quantitative PCR detecting system,qPCR)检测小鼠食管组织葡萄糖转运蛋白-1(Glucose transporters-1,GLUT1)和尿路上皮癌相关基因1(urothelial cancer associated 1,UCA1)mRNA相对表达;蛋白印迹(Western blot,WB)检测小鼠食管组织丙酮酸激酶M2(Pyruvate kinase-M2,PKM2)、磷酸化丙酮酸激酶M2(phospho-PKM2,p-PKM2)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、磷酸化哺乳动物雷帕霉素靶蛋白(phospho-mTOR,p-mTOR)、单羧酸转运蛋白2(monocarboxylate transporter 2,MCT2)、癌基因c-Myc、葡萄糖转运蛋白-1(Glucose transporters-1,GLUT1)蛋白相对表达。结果模型组有癌灶形成,肿瘤细胞突破上皮基底膜,浸润至粘膜下固有层,肿瘤细胞有明显异型性伴角化珠形成,部分呈乳头状,主要以高分化鳞状细胞癌为主,与正常组织之间界限不清;模型组小鼠食管组织α-SMA、Vimentin、PKM2、p-PKM2、MCT2、c-Myc、GLUT1蛋白相对表达和GLUT1 mRNA相对表达显著升高(P<0.05或P<0.01),UCA1 mRNA相对表达降低(P<0.05)。与模型组相比,六君子汤各浓度组小鼠食管癌变程度减轻,六君子汤中浓度食管组织α-SMA、Vimentin、p-PKM2、PKM2、MCT2、mTOR、c-Myc蛋白相对表达显著降低(P<0.05或P<0.01),UCA1 mRNA相对表达显著升高(P<0.05);六君子汤高浓度食管组织α-SMA、Vimentin、p-PKM2、PKM2、MCT2、p-mTOR、mTOR、c-Myc、GLUT1蛋白相对表达和GLUT1 mRNA相对表达显著降低(P<0.05或P<0.01),UCA1 mRNA相对表达显著升高(P<0.01)。结论六君子汤对4NQO诱导的食管癌模型小鼠治疗作用,可能是通过干预肿瘤微环境能量代谢实现的。
Objective To explore the therapeutic mechanism of Liujunzi decoction on esophageal cancer model mice.Methods The mouse model of esophageal cancer was established by 4NQO induction method and divided into blank group,model group,low-concentration group of Liujunzi decoction,medium-concentration group of Liujunzi decoction and high-concentration group of Liujunzi decoction.The protein expressions of α-SMA and Vimentin were detected by immunohistochemistry;The relative expression of GLUT1 and UCA1 mRNA was detected by qPCR;The relative expression of PKM2,p-PKM2,mTOR,p-mTOR,MCT2,c-Myc and GLUT1 proteins were detected by WB.Results Cancer foci were formed in the model group,and tumor cells broke through the epithelial basement membrane and infiltrated into the submucosal lamina propria.Tumor cells showed obvious atypia with keratosis,some of them were papillary,and were mainly highly differentiated squamous cell carcinoma with unclear boundary with normal tissue.Compared with blank group,α-SMA,Vimentin,PKM2,p-PKM2,MCT2,c-Myc,GLUT1 protein relative expression and GLUT1 mRNA relative expression were significantly increased in model group(P<0.05 or P<0.01).Compared with model group,the degree of esophageal cancer was reduced in each concentration groups of Liujunzi decoction,and the relative expressions ofα-SMA,Vimentin,p-PKm2,PKM2,MCT2,mTOR and c-Myc protein in esophagus tissue of medium concentration of Liujunzi decoction were significantly decreased(P<0.05 or P<0.01),the relative expression of UCA1 mRNA was significantly increased(P<0.05);The relative expression ofα-SMA,Vimentin,p-pkm2,PKM2,MCT2,p-mTOR,mTOR,c-Myc,GLUT1 protein and GLUT1 mRNA in high concentration of Lijunzi decoction were significantly decreased(P<0.05 or P<0.01),the relative expression of UCA1 mRNA was significantly increased(P<0.01).Conclusion The therapeutic effect of Liujunzi decoction on 4NQO-induced esophageal cancer model mice may be realized by intervening in the energy metabolism of tumor microenvironment.
作者
尚艺婉
陈星
娄翔宇
武颖烁
周哲旭
刘洋
刘娅茹
胡啸博
菅佳宁
肖小乔
孟丹华
陈玉龙
SHANG Yi-wan;CHEN Xing;LOU Xiang-yu;WU Ying-shuo;ZHOU Zhe-xu;LIU Yang;LIU Ya-ru;HU Xiaobo;JIAN Jia-ning;XIAO Xiao-qiao;MENG Dan-hua;CHEN Yu-long(School of Traditional Chinese Medicine(Zhongjing School),Henan University of Chinese Medicine,Zhengzhou 450046,China;Nanyang Medical College,Nanyang 473000 China;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100000 China;Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046;Henan Key Laboratory of TCM Prescription and Syndrome Signaling,Henan University of Chinese Medicine,Zhengzhou,450046)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2023年第11期2605-2609,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(82074313)。
关键词
食管癌
健脾和胃法
六君子汤
能量代谢
4-硝基喹啉-1-氧化物
Esophageal cancer
Strengthening spleen and harmonizing stomach method
Liujunzi decoction
Energy metabolism
4-nitroquinolin-1-oxide
