摘要
目的观察重楼皂苷Ⅶ(PolyphyllinⅦ,PP7)对斑马鱼HepG2移植瘤和小鼠H;肝癌移植瘤的体内抗肿瘤作用,并探讨其可能的作用机制。方法采用CM-Di I荧光标记的人肝癌Hep G2细胞显微注入斑马鱼卵黄囊内和将小鼠H;肝癌细胞悬液接种到小鼠的右腋窝皮下,建立斑马鱼和小鼠肝癌移植瘤模型。采用不同浓度的PP7(0.06、0.13、0.25、0.50、0.75μmol·L;)处理斑马鱼胚胎;将荷瘤小鼠分为模型组、氟尿嘧啶(5-Fu,25 mg·kg^(-1))组及PP7低、中、高剂量组(0.5、1、2 mg·kg^(-1))。测定给药后斑马鱼移植瘤荧光强度、面积,以及小鼠瘤体质量、体积;统计荷瘤小鼠生命延长率;采用HE染色法对肿瘤组织进行病理形态学观察;采用TUNEL法检测肿瘤细胞凋亡情况;采用免疫组织化学法检测微血管密度(CD31)以及肿瘤细胞增殖(PCNA、Ki67)情况。结果与模型组比较,PP7不同浓度均能够显著降低斑马鱼移植瘤荧光强度和面积(P<0.01);PP7低、中、高剂量均能明显降低小鼠瘤体质量和体积(P<0.05,P<0.01),促进移植瘤细胞凋亡(P<0.05,P<0.01),下调肿瘤组织中CD31、PCNA、Ki67的表达(P<0.05,P<0.01),并且能明显延长荷瘤小鼠生存时间(P<0.05,P<0.01)。结论 PP7具有明显的体内抗肝癌作用,可能与其促进肿瘤细胞凋亡、降低微血管密度以及抑制肿瘤细胞增殖有关。
Objective In this study,the anti-tumor effect of polyphyllinⅦ(PP7) in zebrafish human tumor xenograft model and H;tumor-bearing mice,and the molecular mechanisms were evaluated and explored.Methods We establish zebrafish xenograft model by injecting human hepatocellular carcinoma Hep G2 cells into the yolk sac of zebrafish and H;bearing mice model by subcutaneous inoculating mouse H;hepatocarcinoma cells to mice in the right axillary region.Transplanted zebrafish were treated with different concentrations of PP7(0.06,0.13,0.25,0.50,0.75μmol·L;)and H;bearing mice were divided into five groups and were injected intraperitoneally with PP7(0.5,1,2 mg·kg^(-1)),5-FU(25 mg·kg^(-1))or normal saline(model group).The fluorescence intensity and area of HepG2 cells in zebrafish xenograft model and the tumor weight,tumor size,and life extension in H;bearing mice were monitored.The histological evaluation was corried out by H&E staining of different tumor tissue samples,the immunohistochemical detection of CD31,PCNA and Ki67 in solid tumor sections was performed by standard immunohistochemical techniques,and the percentage of apoptotic cells in tissue section was determined by TUNEL staining.Results Compared with model group,PP7 markedly inhibited the proliferation of Hep G2 cells in zebrafish xenograft model(P<0.01),and decreased the tumor weight and volume at different dosages(P<0.05,P<0.01),induced cancer cell apoptosis of the tumor tissues(P<0.05,P<0.01),decreased the percentage of positive cells expressing PCNA,Ki67 and CD31(P<0.05,P<0.01)in tumor tissue sections,and prolonged the survival period of tumor-bearing mice(P<0.05,P<0.01).Conclusion This study demonstrates that PP7 possesses strong antitumor activity in vivo via suppression of cancer cell proliferation,decrease microvascular density of tumor,and induction of apoptosis in tumor tissues.
作者
张超
黄喜燕
李祥
李庆睿
张栋
李汶芮
张翼
张硕峰
ZHANG Chao;HUANG Xiyan;LI Xiang;LI Qingrui;ZHANG Dong;LI Wenrui;ZHANG Yi;ZHANG Shuofeng(Beijing University of Chinese Medicine,Beijing 100102,China;Beijing Mentougou District Hospital/Capital Medical University Mentougou Teaching Hospital,Beijing 102300,China;Aerospace Central Hospital,Beijing 100049,China;Changchun University of Chinese Medicine,Changchun 130117 Jilin,China;Jilin Agricultural University,Changchun 130117 Jilin,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2022年第1期7-13,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(82003972)
中央高校基本科研业务费专项资助项目(2020-JYB-XJSJJ-011)。
