蒿甲醚抗SD大鼠原位脑胶质瘤血管生成的实验

Inhibitive Effect of Artemether on Brain Glioma Angiogenesis in SD Rat

目的探讨蒿甲醚(Artemether)抗SD大鼠原位脑胶质瘤血管生成作用。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(IC50)。采用立体定位仪在SD大鼠大脑皮质层接种C6脑胶质瘤细胞(1×106/μl)40只,雌、雄各半;随机分为5组,每组8只。在接种第3天后,各组采用灌胃给药法连续给药10天。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)。全脑标本用4%多聚甲醛固定,肿瘤组织做病理观察,免疫组化方法检测移植瘤组织微血管密度。结果各实验组血管计数分别为Ⅰ组(39±4),Ⅱ组(29±6),Ⅲ组(12±8),Ⅳ组(10±5),生理盐水组为(52±7)。各实验组血管计数均明显少于生理盐水对照组,差异有统计学意义(分别P<0.05,P<0.01)。各实验组SD大鼠原位脑胶质瘤体积较对照组显著减小。结论在一定剂量范围内,蒿甲醚具有明显抑制SD大鼠原位脑胶质瘤血管生成作用;蒿甲醚抑制原位脑胶质瘤生长和转移的机制之一是透过血脑屏障抑制脑胶质瘤血管生成。

Objective To investigate inhibitory effect of Artmeter （Artm）on brain glioma angiogenesis in SD rat. Methods MTT （Methy thiazolyl tetrazolium） assay was used to evaluate the inhibitory effect of Artemether treatment on C6 glioma cells. Forty （20 male and 20 female） SD rats which was subcutaneous implantation with SD rat C6 glioma cell （1×10^6/μl） in cerebral cortex were divided into 5 groups randomly, each group had 8 rats. All were taken orally. After 3 day of implantation the treatment was started and continued until 10^th day, and the rat killed 20^th day. Whole brain was fixed and length-path （a ram） and short-path （b mm） of tumors were measured. Volume of tumor was calculated following formula： V （mm3） = a^2bπ / 6. Micro-vascular dense （MVD） was observed under the microscopy by im munohistochemistry. Results The value of Micro-vascular dense in different therapy groups was group Ⅰ （39 ± 4）, group Ⅱ（29 ± 6）, group Ⅲ （12 ± 8）and group Ⅳ （10 ± 5）, normal saline control group is （52 ± 7）, respectively, Micro-vascular dense （MVD） in different therapy groups was significant lower than that in control group （P〈0. 05, P〈0. 01 respectively） and volume of brain glioma in different therapy groups is significant smaller than that in control group （P〈0. 05, P〈0. 01 respectively）. Conclusion There is significant inhibitory effect of Artmeter on brain glioma angiogenesis in SD rat in some arrange dosage. The mechanism that Artemether inhibits brain glioma growth and metastasis may be penetrating the blood-brain barrier by inhibiting angiogenesis.