蒿甲醚对大鼠原位脑胶质瘤抑瘤及抗血管生成的实验研究

目的探讨蒿甲醚是否在选择性地杀伤SD大鼠原位脑胶质瘤的同时还具有抑制血管生成的作用.方法采用四甲基偶氮唑蓝(MTT)法测定不同浓度蒿甲醚对大鼠C6脑胶质瘤细胞株的生长抑制作用,计算半数抑制浓度(IC5)0;采用立体定位仪在48只SD大鼠大脑皮质层接种C6脑胶质瘤细胞(1×106个/μL),雌、雄各半;随机分为3组,每组8只.分别为空白对照组、阳性对照和实验组.在接种第3天后,实验组各组采用灌胃给药法连续给药10 d;于接种后的第20天解剖大鼠,经活体左心室灌注4%多聚甲醛,固定肿瘤的全脑标本.在大鼠脑部接种穿刺点做冠状切口,按垂直和水平方向测量肿瘤大小.肿瘤体积=a2bπ/6(a为肿瘤的短径,b为肿瘤的长径).采用免疫组化方法检测移植瘤组织微血管密度.结果实验组各组对SD大鼠原位脑胶质瘤的抑瘤率分别为:54.5%、61.0%、64.5%和69.8%,与空白对照组比较,差异有统计学意义(P<0.01);各实验组血管计数均明显少于空白对照组,差异有统计学意义(P<0.05、P<0.01).各实验组SD大鼠原位脑胶质瘤体积较空白对照组显著减小.结论在一定剂量范围内,口服蒿甲醚对SD大鼠脑部原位接种C6脑胶质瘤有明显的抑制作用;蒿甲醚抑制原位脑胶质瘤生长的机制之一可能是透过血脑屏障抑制脑胶质瘤血管生成.

Study on Inhibitory Effects of Artemether on Brain Glioma Growth and Angiogenesis in SD Rats

Objective To explore the inhibitory effect of artemether(Artm)on glioma growth and angiogenesis in brain tumor bearing SD rats.Methods MTT(Methy thiazolyl tetrazolium) assay was used to evaluate the inhibitory effect of Artemether treatment on C6 glioma cells.48(24 male and 24 female)SD rats which were subcutaneous planted with SD rat C6 glioma cell(1×106/μL) in cerebral cortex were divided randomly into 3 groups,each group had 8 rats.Blank control group:rats were orally given normal saline.Positive control group: rats were orally given Lomustine.Different artemether therapy groups: rats were given different doses of artemether.Treatment started on day 3 and continued until day 10 and SD rats were killed on day 20.Whole brain was fixed with 4% paraformaldehyde through alive left ventricle perfusion.The length-path and short-path of tumor each rat was measured by staff gauge in vertical and horizontal.Volume of tumor was calculated following formula: V(mm3) = a2bπ6(a:short-path,b:length-path).Micro-vascular dense(MVD)was observed and countered under the microscopy by immunohistory chemistry.Results Inhibitory rates of Artemether at different dosages were 54.5%,61.0%,64.5% and 69.8%,respectively,and compared with blank control group,different experiment groups had significant difference(P=0.000 respectively).Micro-vascular dense in different therapy groups was significant lower than that in blank control group(P<0.05,P<0.01 respectively)and volume of brain glioma in different therapy groups was significant smaller than that in blank control group.Conclusions Artemether has inhibitory effect on brain glioma growth in SD rats.The mechanism of Artemether inhibiting brain glioma growth may be penetrating the blood-brain barrier by inhibiting angiogenesis.