壁虎提取物.doc

壁虎提取物抗小鼠H22肝癌及免疫调节 罗文军 王建刚★ （河南科技大学医学院药理教研室，洛阳，471003’s immunological mechanisms of Gekko Extract(GE).Methods H22 tumor-bearing model KM mice were applied by intraperitoneal injection to evaluated in vivo antitumor activity and immunoregulatory effect of GE. The influence of different dose GE(0.4,0.8,1.6g/kg) on immune organs and antitumor effects in tumor-bearing mice were observed.The levels of IL-12 and IL-18 in serum from mice were detected by means of ELISA Kits. Results GE possessed markedly inhibitory activity on the growth of H22 tumor in mice,the dosage of gekko extract were1.60，0.80 or 0.40 mg·Kg-1, theirs inhibitory rates were 55.25％、47.89％and l5.16％respectively with H22 liver cancer. To compare with model group、Positive group，theirs difference was obvious,and had distinct effect on weight of thymus and spleen was observed,peripheral blood white blood cells were proliferated,the contents of serum IL-12/IL-18 were induced significantly. Conclusion GE not only shows obvious antitumor effect,but also protects thymus and spleen, GE(s Immunoregulation can Induce WBC,IL-12 and IL-18 production KEYWORDS: GE; antitumor; immunoregulation 壁虎作为我国传统中药，具有补肺肾、益精血、止咳定喘、镇痉祛风和发散消肿的功效。,ylwjg@163.com 1材料和方法 1.1材料 1.1.1实验动物 昆明(KM)小白鼠50只，雄性，6周龄，体重18－20克，由河南科技大学实验动物研究中心提供。H22瘤株，由河南医科所提供。 1.1.2材料和试剂 小鼠IL-12，IL-18、ELISA试剂盒购自美国BioSource公司，酶联测定仪（Biocell-2010，郑州博赛生物工程有限公司），阿霉素，浙江海正药业股份有限公司产品批号国药准字 1.1.3.GE制备：壁虎购自安徽省毫州市永刚饮片厂有限公司，放置-20℃冰冻2h，取出待恢复室温后再放置-20℃冰冻2h再取出，加入含有50mmol/lNaCL的20mmol/l Tris-HCL，用高速组织捣碎机使细胞裂解，然后8000r/m 4℃离心30min，弃上清，沉淀用Tris-HCl缓冲液洗涤数次，加入20mmol/lHCl,均质，8000r/m 4℃30min离心收集上清，沉淀用20mmol/lHCl再抽提2次，3次收集的上清液40℃旋转蒸发冷冻干燥浓缩成干粉低温保存，备用。 方法1.2.1小鼠肝癌H22模型的建立： 将冻存H22细胞复苏，传代，以无菌生理盐水将处于对数生长期H22 细胞 ，调整细胞浓度为1.0×107/ml， 取0.2ml注入昆明小鼠腹腔，6-8天成腹水，无菌条件下用5ml注射器抽吸除1～2ml牛奶状、较粘稠的腹水，先用0.4％台盼兰染色，计数活细胞率90％，再用灭菌0.9%生理盐水稀释调整浓度为1×107/ml，每只小鼠于右前肢腋窝皮下接种0.2ml，制成实体型荷瘤鼠模型，30分钟内完成。 1.2.2分组、药物配制及给药方法： 将50只小鼠荷瘤小鼠随机分为5组:模型组、ADM组、GE高中低组，每组各10只称重、标记并记录。ADM组，以5mg/kg剂量配２ｍｇＡＤＭ溶于８ｍｌ无菌水的比例配制，