山慈菇多糖对肝癌腹水荷瘤小鼠症状的改善机制  

Improvement mechanism of Pseudobulbus Cremastrae Seu Pleiones polysaccharide on the symptoms of hepatoma ascites bearing mice

作　　者：张志强[1] 魏毅强 杨亚莉[1] 张晓菲 ZHANG Zhiqiang;WEI Yiqiang;YANG Yali;ZHANG Xiaofei(No.1 Department of Gastroenterology,Xinxiang First People's Hospital,Xinxiang 453000,China;Department of Gastroenterology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]新乡市第一人民医院消化内科一病区,新乡453000 [2]郑州大学第一附属医院消化内科,郑州450000

出　　处：《西北药学杂志》2023年第3期74-80,共7页Northwest Pharmaceutical Journal

基　　金：2019年河南省医学科技攻关计划(编号:LHGJ20190022)。

摘　　要：目的 探讨山慈菇多糖对肝癌腹水荷瘤小鼠症状的改善作用并阐明其可能的机制。方法 于55只C57BL/6雄性小鼠中随机选择5只腹腔注射肝癌腹水型H_(22)细胞,随机选择10只作为正常组,剩余小鼠通过腹水接种诱导肝癌腹水荷瘤小鼠模型。成功建模后,随机分为模型组、山慈菇多糖低剂量(200 mg·kg^(-1)·d^(-1))、高剂量组(400 mg·kg^(-1)·d^(-1))及环磷酰胺组(20 mg·kg^(-1)·d^(-1)),每组各10只,正常组及模型组小鼠均腹腔注射等体积生理盐水0.2 mL,1天1次,连续14 d。末次给药结束后,观察小鼠一般体征并称定质量;记录小鼠腹水体积;检测外周血红细胞(RBC)、白细胞(WBC)总数;处死后,剥离肿瘤称定质量,计算抑瘤率;摘取脾脏、胸腺称定质量,计算脏器指数;HE染色观察肿瘤组织学变化;TUNEL染色检测肿瘤细胞凋亡情况;免疫组化技术检测肿瘤组织中血小板内皮细胞粘附分子1(CD31)抗原表达情况;Western blot法检测肿瘤组织中磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白PI3K、p-PI3K、AKT、p-AKT、mTOR、p-mTOR、Beclin1、B细胞淋巴瘤/白血病-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)及血管内皮生长因子(VEGF)蛋白相对表达情况。结果 成功构建肝癌腹水荷瘤小鼠模型;与正常组比较,模型组小鼠腹胀、消瘦及皮肤暗沉发黄,被毛无光泽;血性腹水量多,外周血RBC总数减少,WBC总数增加,肿瘤体质量增加(P<0.05),免疫器官脾脏及胸腺指数均降低(P<0.05),肿瘤细胞生长良好,异型性高,周围血管丰富,分布密集,且肿瘤组织中mTOR通路相关蛋白p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR、Bcl-2及VEGF蛋白相对表达水平升高,Beclin1及Bax蛋白相对表达水平降低(P<0.05);与模型组比较,山慈菇多糖2组及环磷酰胺组小鼠临床症状有所改善,腹水减少,外周血RBC总数升高,WBC总数降低,肿瘤体质量减少(P<0.05),抑瘤率分别为25.31%、45Objective To investigate the effect of Pseudobulbus Cremastrae Seu Pleiones polysaccharide on the symptoms of mice bearing hepatoma ascites and elucidate its possible mechanism.Methods 5 of 55 male C57BL/6 mice were randomly selected for intraperitoneal injection of hepatocellular carcinoma ascites type H22 cells,and 10 mice were randomly selected as the normal group.The rest mice were inoculated with ascites to induce hepatocellular carcinoma ascites bearing mice model.After successful modeling,the mice were randomly divided into model group,Pseudobulbus Cremastrae Seu Pleiones polysaccharide low dose group(200 mg·kg^(-1)·d^(-1)),high dose group(400 mg·kg^(-1)·d^(-1))and cyclophosphamide group(20 mg·kg^(-1)·d^(-1)),with 10 mice in each group.The normal group and model group were intraperitoneally injected with 0.2 mL normal saline,once a day,for 14 consecutive days.After the last administration,the general physical signs of mice and weigh the weight were observed.The volume of ascites in mice was recorded.The total number of red blood cells(RBC)and white blood cells(WBC)in peripheral blood was measured.After being killed,the tumor was stripped and weighed,and the tumor inhibition rate was calculated.The spleen and thymus were extracted and weighed,and the viscera index was calculated.HE staining was used to observe the histological changes of tumor.TUNEL staining was used to detect the apoptosis of tumor cells.Immunohistochemical technique was used to detect the expression of platelet endothelial cell adhesion molecule 1(CD31)antigen in tumor tissue.Western blot was used to detect the relative expression of phosphatidylinositol-3 kinase(PI3K)/protein kinase B(AKT)/mammalian rapamycin target protein(mTOR)pathway related proteins PI3K,p-PI3K,AKT,p-AKT,mTOR,p-mTOR,Beclin1,B-cell lymphoma/leukemia 2 gene(Bcl-2),Bcl-2 related X protein(Bax)and vascular endothelial growth factor(VEGF)protein in tumor tissue.Results The tumor-bearing mouse model of hepatocellular carcinoma ascites was successfully constructed.Com

关 键 词：山慈菇多糖 肝癌 哺乳动物雷帕霉素靶蛋白通路 

分 类 号：R965[医药卫生—药理学]