毛钩藤碱联合奥沙利铂对胃癌HGC-27荷瘤小鼠的抗肿瘤作用研究  

Study on the antitumor effect of hirsutine combined with oxaliplatin on HGC-27 tumor-bearing gastric cancer mice

作　　者：万如意 周波[2] 陈博[2] Wan Ruyi;Zhou Bo;Chen Bo(Anhui Medical University First Clinical School,Hefei 230022;Dept of General Surgery,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学第一临床医学院,合肥230022 [2]安徽医科大学第一附属医院普外科,合肥230022

出　　处：《安徽医科大学学报》2024年第12期2150-2156,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金项目(编号:2208085MH240)。

摘　　要：目的探讨毛钩藤碱(hirsutine)联合奥沙利铂(oxaliplatin)对胃癌HGC-27荷瘤小鼠抗肿瘤作用的影响。方法构建胃癌HGC-27荷瘤小鼠模型,将建模成功的荷瘤小鼠随机分为正常对照组(normal)、模型组(model)、oxaliplatin组(10 mg/kg)、hirsutine组(10 mg/kg)和联合组(combination)(hirsutine 10 mg/kg+oxaliplatin 10 mg/kg),连续干预2周。记录各组小鼠肿瘤生长情况,计算抑瘤率、胸腺指数和脾脏指数;流式细胞术检测外周血中CD4^(+)T细胞和CD8^(+)T细胞亚群比例,并计算CD4^(+)/CD8^(+)比值;ELISA检测血清中白细胞介素(IL)-1β、IL-6和肿瘤坏死因子(TNF)-α水平;HE染色观察肿瘤组织病理学变化;Western blot法检测肿瘤组织中活化胱天蛋白酶3(cleaved Caspase-3)、Bcl-2关联X蛋白(Bax)和B淋巴细胞瘤-2(Bcl-2)以及肿瘤增殖抗原Ki67、增殖细胞核抗原PCNA蛋白表达水平。结果与normal组比较,model组小鼠脾脏指数、胸腺指数以及外周血CD4^(+)T细胞比例和CD4^(+)/CD8^(+)比值均降低(P<0.05),而外周血CD8^(+)T细胞比例、血清IL-1β、IL-6和TNF-α含量增加(P<0.05);与model组比较,oxaliplatin组、hirsutine组、combination组3组小鼠肿瘤生长均较为缓慢,肿瘤组织细胞体积减小,异型性较小,细胞核固缩减少,同时抑瘤率、脾脏指数、胸腺指数以及外周血CD4^(+)T细胞比例和CD4^(+)/CD8^(+)比值均增加(P<0.05),外周血CD8^(+)T细胞比例、血清IL-1β、IL-6和TNF-α含量均减少(P<0.05),肿瘤组织中Bcl-2、Ki67、PCNA蛋白表达水平均降低,cleaved Caspase-3和Bax蛋白表达水平均升高(P<0.05),其中以毛钩藤碱与奥沙利铂联合使用的combination组干预效果最为明显。结论毛钩藤碱可抑制胃癌荷瘤小鼠肿瘤细胞的增殖,促进其凋亡,且与奥沙利铂联合使用效果更为显著。Objective To investigate the effect of hirsutine combined with oxaliplatin on the antitumor activity of HGC-27 mice with gastric cancer.Methods Gastric cancer HGC-27 tumor-bearing mice model was established.The successfully established tumor-bearing mice were randomly divided into normal control group,model group,oxaliplatin group(10 mg/kg),hirsutine group(10 mg/kg)and combination group(10 mg/kg hirsutine+oxaliplatin 10 mg/kg).The intervention lasted for 2 weeks.The tumor growth of mice in each group was recorded,and the tumor inhibition rate,thymus index and spleen index were calculated.The proportion of CD4^(+)T cells and CD8^(+)T cell subsets in peripheral blood was detected by flow cytometry,and the CD4^(+)/CD8^(+)ratio was calculated.ELISA was used to measure the serum levels of interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α.HE staining was used to observe the pathological changes in tumor tissues.Western blot was used to detect the protein expression levels of cleaved Caspase-3,Bcl-2 associated X protein(Bax),B-cell lymphoma-2(Bcl-2)and tumor proliferation antigen(Ki67),proliferating cell nuclear antigen(PCNA)in tumor tissues.Results Compared with the control group,the spleen index,thymus index,CD4^(+)T cell ratio and CD4^(+)/CD8^(+)ratio in peripheral blood of the model group decreased(P<0.05),while the proportion of CD8^(+)T cells in peripheral blood and the levels of TNF-α,IL-1βand IL-6 in serum increased(P<0.05).Compared with the model group,the tumor growth in oxaliplatin group,hirsutine group and combined group was significantly inhibited,the volume of tumor cells decreased,the heteromorphism was smaller,and the nucleosomes decreased less.The inhibitory rate,spleen index,thymus index,CD4^(+)T cell ratio and CD4^(+)/CD8^(+)ratio in peripheral blood of the model group increased(P<0.05),the proportion of CD8^(+)T cells in peripheral blood and the levels of TNF-α,IL-1βand IL-6 in serum decreased(P<0.05),while the protein expression levels of Bcl-2,Ki67 and PCNA in tumor tissues decreased a

关 键 词：毛钩藤碱 奥沙利铂 HGC-27 抗肿瘤 胃癌 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]