越鞠丸干预DMBA诱导乳腺癌模型大鼠的效应及机制研究  

Effect and mechanism of Yueju Pills on breast cancer induced by DMBA in rats

作　　者：胡文秀 王馨楠 刘祯宏 赵益[4] 孙有智[1,2] HU Wen-xiu;WANG Xin-nan;LIU Zhen-hong;ZHAO Yi;SUN You-zhi(Formula-Pattern Research Center,Jiangxi University of Chinese Medicine,Nanchang 330004,China;School of Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004,China;School of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Research Center of Differential and Development of Traditional Chinese Medicine Basic Theory,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学方证研究中心,江西南昌330004 [2]江西中医药大学中医学院,江西南昌330004 [3]江西中医药大学药学院,江西南昌330004 [4]江西中医药大学中医基础理论分化发展研究中心,江西南昌330004

出　　处：《中国中药杂志》2024年第2期431-442,共12页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82060868,82360961);江西省教育厅科技项目(GJJ2200907);江西省中医药骨干人才培养项目(赣中医药科教字[2021]4号);江西中医药大学校级科技创新团队发展计划项目(CXTD22007);熊墨年国医名师传承工作室建设项目(赣中医药综合字[2021]12号);江西省高等学校大学生创新创业训练计划项目(202210412323)。

摘　　要：探究越鞠丸对乳腺癌的抑制作用及其机制。92只SPF级SD雌性大鼠随机选14只为对照组;其余78只采用7,12-二甲基苯并蒽(DMBA)建立乳腺癌模型。将模型建立成功的大鼠随机分为模型组、他莫昔芬组(1.9 mg·kg^(-1)·d^(-1))、越鞠丸低剂量组(17 g·kg^(-1)·d^(-1))、越鞠丸高剂量组(34 g·kg^(-1)·d^(-1))。每日观察大鼠精神、饮食、活动,隔日称量体质量,给药12周后处死动物,称量瘤重;酶联免疫吸附实验(ELISA)检测血清雌激素、孕酮水平;HE染色观察乳腺和肿瘤组织病理学改变;蛋白质免疫印迹法(Western blot)测定葡萄糖转运蛋白1(glucose transporter 1,GLUT1)、乳酸脱氢酶A(lactate dehydrogenase A,LDHA)、磷酸果糖激酶(phosphofructokinase muscle,PFKM)、丙酮酸激酶M2(pyruvate kinase isozyme type M2,PKM2)、己糖激酶2(hexokinase 2,HK2)和核因子-κB(nuclear facter-κB,NF-κB)、磷酸化NF-κB表达情况;采用16S rRNA高通量测序检测肠道微生物组。结果显示,与模型组相比,越鞠丸高、低剂量组肿瘤抑制率分别为15.8%、64.5%,低剂量组抑制作用更为显著;与对照组相比,模型组血清中雌激素、孕酮水平明显升高,越鞠丸给药后血清中雌激素、孕酮水平相较于模型组呈下降趋势,且越鞠丸低剂量组作用显著(P<0.05);HE染色结果显示,与模型组比较,越鞠丸给药组乳腺组织增多的腺体减少,乳腺导管扩张程度减轻,腺泡数量与腺泡腔面积减少,分泌物减少,乳腺上皮细胞的复层排列层次减少;同时研究发现越鞠丸干预后糖酵解相关蛋白GLUT1、LDHA、PFKM、PKM2、HK2和炎症因子NF-κB的表达显著下调(P<0.05),并使乳腺癌大鼠肠道微生物群落多样性、微生物组成和结构及群落丰度发生改变。研究结果表明,越鞠丸对乳腺癌具有抑制作用,其作用机制可能与调节雌激素、孕激素分泌,调控糖酵解和炎症因子以及影响肠道微生物组成密切相关。This paper aims to explore the inhibitory effect of Yueju Pills on breast cancer and decipher the underlying mechanism.A total of 92 SPF-grade SD female rats were involved in this study,and 14 of them were randomly selected into control group.The remaining 78 rats were administrated with 7,12-dimethylbenzanthracene(DMBA) by gavage to establish the breast cancer model.The modeled rats were randomized into model,tamoxifen(1.9 mg·kg~(-1)·d~(-1)),and low-and high-dose(17,34 g·kg~(-1)·d~(-1)) Yueju Pills groups.The mental state,food intake,and activities of the rats were observed daily,and the body weight was measured on alternate days.After 12 weeks of administration,the rats were sacrificed and the tumor weight was measured.The serum estrogen and progeste-rone levels were determined by enzyme-linked immunosorbent assay.The histopathological changes of the breast and tumor were observed by hematoxylin-eosin staining.Western blot was employed to measure the protein levels of glucose transporter 1(GLUT1),lactate dehydrogenase A(LDHA),phosphofructokinase muscle(PFKM),pyruvate kinase isozyme type M2(PKM2),hexokinase 2(HK2),nuclear factor-kappaB(NF-κB),and phosphorylated NF-κB.The intestinal microbiome was examined by 16S rRNA high-throughput sequencing.The results showed that compared with the model group,high and low-dose Yueju Pills showed the tumor inhibition rate of 15.8% and 64.5%,respectively,and the low dose group had stronger inhibitory effect.Compared with the control group,the model group presented elevated the levels of estrogen and progesterone in serum.The administration of Yueju Pills lowered such ele-vation,and the low-dose group showed stronger lowering effect(P<0.05).Compared with the model group,Yueju Pills reduced the glands with increased breast tissue,the degree of breast duct expansion,the number and area of acinar cavity,the secretions,and the layers of mammary epithelial cells.Furthermore,Yueju Pills down-regulated the expression of GLUT1,LDHA,PFKM,PKM2,HK2,and NF-κB(P<0.05) and altered the

关 键 词：越鞠丸 乳腺癌 糖酵解 炎症因子 肠道菌群 

分 类 号：R285.5[医药卫生—中药学]