小鼠肿瘤细胞 枸杞多糖对实验性肝癌小鼠肿瘤细胞FasL表达的影

枸杞多糖对实验性肝癌小鼠肿瘤细胞FasL表达的影

【结果】 lbp低、高剂量组均可显著降低肿瘤细胞核分裂计数(p＜0  05)；
lbp高 剂量组可升高肿瘤间质淋巴细胞浸润程度(p＜0  05),he染色显示在肿瘤边缘组 织内见多量淋巴细胞浸润。lbp低、高剂量组均可显著降低小鼠肿瘤细胞fasl阳性 表达指数(p＜0  05)；
免疫组化染色显示模型组fasl表达强阳性,lbp低剂量组部分 肝癌细胞内fasl表达阳性,fasl仅在lbp高剂量组少量肝癌细胞中有表达。【结论】 lbp抗实验性肝癌的效应可能与其能抑制fasl表达,减少免疫活性细胞凋亡有关。

Www.133229.COM 【关键词】 枸杞多糖/药理学 肝肿瘤/中药疗法 肿瘤组织/病理学 基因表 达调控 疾病模型,动物 小鼠 abstract: objectiveto observe the influence of lycium barbarum polysaccharide(lbp) on fasl expression in h22  bearing mice and to explore its anti  tumor mechanism. methodskunming mice were randomized into the model group, and low  and high  dose lbp (in the dose of 0  625 and 1  250 g·kg-1·d-1 respectively) groups. h22  bearing mice models were induced through right subaxillary inoculation of h22  ascitic cells. three days after inoculation, lbp group were given lbp for 14 days. hematoxylin and eosin(he) staining method was used to examine the tumor cell density, tumor cell mitotic count, and lymphocyte infiltration in tumor interstitial tissue. fasl expression was observed in the three groups with immunohistochemical method. resultstumor cell mitotic count was decreased in the two lbp groups (p0  05), the score of lymphocyte infiltration in tumor interstitial tissue was increased,and lymphocyte infiltration was obvious after he staining in high  dose lbp group (p0  05). the index of fasl expression positive was decreased in both lbp groups (p0  05).fasl expression was strong positive in the model group, and positive in partial liver tumor cells of low  dose lbp group and in less liver tumor cells of high  dose lbp group. conclusionthe anti  tumor mechanism of lbp is probably related with the inhibition of fasl expression and with the suppression of apoptosis of immune active lymphocytes． key words:lycium barbarum polysaccharide/pharmacology;liverneoplasms/tcd therapy;neoplasms /pathology;gene expression regulation;disease models, animal;mice 死亡分子fas(apo  1/cd95)和其配体fasl相互作用是细胞凋亡的重要途 径之一。肿瘤细胞可通过表达fasl,与活化的fas(+)的淋巴细胞接触,主动杀伤免疫 活性细胞,实现其免疫逃逸,故又称为fas反击［1-2］。研究证实枸杞多糖(lbp)具有 广泛的抗肿瘤效应［3-4］,本研究通过观察小鼠实验性肝癌组织fasl表达情况及病 理学改变,探讨lbp抗实验性肝癌作用可能存在的机制和靶点。现报道如下。

1 材料与方法 1.1 动物与瘤株昆明种小鼠,清洁级,6～8周龄,体质量(18±2)g,雌雄各 半,由广州中医药大学实验动物中心提供,合格证号：0002769；
h22肝癌腹水型细 胞株由中山大学医学院实验动物中心细胞库提供。

1.2 药物与试剂枸杞多糖(lycium barbarum polysaccharide,lbp)由广州 中医药大学热带医学研究所药物化学研究室从优质宁夏枸杞子中分离提取,采用 双光束紫外可见分光光度计测定lbp含量为35  8 g/kg；
兔抗fasl免疫组化试剂为 boster biotechnology公司产品,批号：ba0049；
兔多克隆抗体免疫组织化学染色试 剂盒(sa1022)、二氨基联苯胺(3,3′  diaminobenzidine,dab)显色试剂盒均购自武汉 博士德生物工程有限公司；
甲醛、乙醇、二甲苯均为汕头市光华化学厂产品；
石 蜡为江苏太仓皓天石蜡厂产品。

1.3 仪器tu  1901双光束紫外可见分光光度计(北京普析通用仪器有 限责任公司)；
libror aeg  220电子天平(日本岛津)；
nex power 1000型纯水器(韩 国human corp)；
to1020脱水机、eg1160包埋机、rm2135轮转切片机、st4040染色 机均购自德国leica公司；
cx21显微镜(日本olympus)。