姜黄素和姜黄素联氨基抗肝癌作用比较及机制研究  

Comparison of Anti-hepatocarcinoma Effect of Curcumin and Hydrazincurcumin and Mechanism Study

作　　者：赵冀安[1] 崔丽敏[2] 董梁[3] 聂文佳[3] 刘文聪[4] 李增宁[5] ZHAO Ji’an;CUI Limin;DONG Liang;NIE Wenjia;LIU Wencong;LI Zengning(Dept.of General Surgery,the First Hospital of Hebei Medical University,Shijiazhuang 050013,China;Dept.of Medical Oncology,Dingzhou Municipal People’s Hospital,Hebei Dingzhou 071010,China;Dept.of Medical Service,the First Hospital of Hebei Medical University,Shijiazhuang 050013,China;Dept.of Ultrasonography,the First Hospital of Hebei Medical University,Shijiazhuang 050013,China;Dept.of Clinical Nutrition,the First Hospital of Hebei Medical University,Shijiazhuang 050013,China)

机构地区：[1]河北医科大学第一医院普通外科,石家庄050013 [2]定州市人民医院肿瘤内科,河北定州071010 [3]河北医科大学第一医院医务处,石家庄050013 [4]河北医科大学第一医院超声科,石家庄050013 [5]河北医科大学第一医院临床营养科,石家庄050013

出　　处：《中国药房》2020年第22期2741-2750,共10页China Pharmacy

基　　金：河北省医学科学研究课题计划(No.20190485)。

摘　　要：目的:比较姜黄素(CUR)和其衍生物联氨基姜黄素(HZC)的体内外抗肝癌作用及机制。方法:采用MTT法检测CUR或HZC(2.5、5、10、20、40、80μmol/L)对人肝癌HepG2细胞增殖的影响;采用流式细胞术检测CUR或HZC(10、20、40μmol/L)对HepG2细胞周期分布和凋亡情况的影响;采用Western blotting法检测CUR或HZC(10、20、40μmol/L)对HepG2细胞凋亡相关蛋白表达的影响。将雄性SD大鼠随机分为正常对照组(n=10)、CUR对照组(n=10)、HZC对照组(n=10)、肝癌模型组(n=30)、CUR防护组(n=30)、HZC防护组(n=30)。CUR对照组和HZC对照组大鼠分别腹腔注射CUR或HZC(剂量均为80 mg/kg);模型组、CUR防护组和HZC防护组大鼠均腹腔注射对二乙基亚硝胺(50 mg/kg)建立肝癌模型,同时2种药物防护组大鼠分别腹腔注射CUR或HZC(剂量均为80 mg/kg),每周2次,连续给药12周。给药期间,记录大鼠体质量变化和死亡情况;24周后,计算大鼠肝脏指数并观察其外观,统计肝癌结节数;采用苏木精-伊红染色法观察大鼠肝组织病理学变化并计算肝癌细胞核分裂指数;采用免疫组化法检测大鼠肝组织增殖细胞核抗原(PCNA)指数。结果:CUR和HZC均能显著升高HepG2细胞的增殖抑制率、G0/G1期细胞周期百分比和凋亡率(P<0.05);均能显著降低细胞中磷酸化蛋白酪氨酸激酶2(p-JAK2)、磷酸化信号转导及转录激活蛋白3(p-STAT3)、B淋巴细胞瘤2(Bcl-2)、Bcl-xl蛋白的表达水平,显著升高Bcl-2相关X蛋白(Bax)、细胞色素C(Cyt-c)、胱天蛋白酶9(Caspase-9)、Caspase-3、聚腺苷二磷酸核糖聚合酶(PAPR)蛋白的表达水平(P<0.05);且HZC的上述作用均显著强于CUR(P<0.05)。动物实验结果显示,3个对照组大鼠无死亡,未发生肝脏癌变,肝脏外观和组织切片未见病理变化;体质量及增量、肝脏指数、癌细胞核分裂指数、肝组织PCNA指数组间比较差异均均无统计学意义(P>0.05)。与肝癌模型组比较,CUR防护组和HZC防护组大鼠的存OBJECTIVE:To compare the anti-hepatocarcinoma effects of curcumin(CUR)and its derivative hydrazincurcumin(HZC),and to explore the mechanism.METHODS:MTT assay was used to detect the effects of CUR or HZC(2.5,5,10,20,40,80μmol/L)on the proliferation of HepG2 cells.Flow cytometry was used to detect the effects of CUR or HZC(10,20,40μmol/L)on cell cycle distribution and apoptosis of HepG2 cells.Western blotting assay was used to detect the effects of CUR or HZC(10,20,40μmol/L)on the expression of apoptosis-related protein in HepG2 cells.The male SD rats were randomly divided into normal control group(n=10),CUR control group(n=10),HZC control group(n=10),model group(n=30),CUR protection group(n=30)and HZC protection group(n=30).CUR control group and HZC control group were given CUR or HZC(80 mg/kg)intraperitoneally.Model group,CUR protection group and HZC protection group were given diethylnitrosamine(50 mg/kg)intraperitoneally to establish hepatocarcinoma model;at the same time,2 protection groups were given CUR or HZC(80 mg/kg)intraperitoneally,twice a day,for consecutive 12 weeks.During medication,the change of body weight and death of rats were recorded.Twenty four weeks later,liver index of rats was calculated and appearance was observed;the number of cancer nodules was counted;HE staining was used to observe the pathological changes of liver tissue and calculate the nuclear division index of hepatocarcinoma;the proliferating cell nuclear antigen(PCNA)index was detected by immunohistochemistry.RESULTS:CUR and HZC could increase the inhibitory rate of HepG2 cells(P<0.05),and increased the percentage at G0/G1 phase and apoptotic rate of HepG2 cells(P<0.05).CUR and HZC could significantly decrease the protein expression of p-JAK2,p-STAT3,Bcl-2 and Bcl-xl,while increased the protein expression of Bax,Cyt-c,Caspase-9,Caspase-3 and PAPR(P<0.05).Above effects of HZC were significantly better than those of CUR(P<0.05).The results of animal experiment showed that there was no death,no liver canceration and no pathologi

关 键 词：肝癌 姜黄素 联氨基姜黄素 蛋白酪氨酸激酶2/信号转导及转录激活蛋白3信号通路 HEPG2细胞 大鼠 核分裂指数 增殖细胞核抗原指数 

分 类 号：R735.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]