尖吻蝮蛇小分子多肽对U251荷瘤鼠体内抑制实验研究

The study of polypeptide from agkistrodon acutus venom inhibitory effects on U251 glioma subcutaneous model in nude mice

目的:研究尖吻蝮蛇小分子多肽对U251荷瘤鼠的体内抑制作用及瘤细胞增殖活性变化,并初步探讨其作用机制。方法:取生长状态良好的U251胶质瘤细胞悬液按1×106个细胞/50μl接种于荷瘤鼠腹腔皮下,待其成瘤后将肿瘤组织块接种于只实验鼠,造模成功的60只大鼠随机分为实验组、顺铂组及空白对照组,于治疗7d后计算瘤体变化,通过光镜、电镜等方法,观察小分子多肽对胶质瘤作用的形态学变化,并应用免疫组织化学方法,检测增殖细胞核抗原(PCNA)的表达,观察瘤细胞增殖变化情况。结果:实验组蛇毒小分子多肽在30μg/kg剂量时对U251胶质瘤细胞有显著抑制作用,60μg/kg剂量抑瘤率可达53.37%。实验组肿瘤内PCNA阳性细胞数量较对照组显著下降。结论:蛇毒小分子多肽能够抑制体内U251胶质瘤细胞的生长;其作用机制可能与诱导脑胶质瘤细胞发生凋亡及抑制肿瘤细胞增殖有关。

Objective：To study the inhibiting effect on the growth and proliferated activity of the U251 glioma subcutaneous model in nude mice and after polypeptide from agkistrodon acutus venom treatment.Methods：Well cultured U251 a glioma cell suspension was inoculated into the inguinal groove subcutaneous tissue of nude mice by 10^6 cells/50μl,and when tumors appeared,another nude mice were inoculated with the formed tumor tissue pieces.60 model mice were randomly divided to experiment group,Cisplatin group and control group.The size of glioma were measured,by which we calculate inhibitory rate.Proliferation and morphological change of glioma cells were detected by HE staining method,electron-microscopic observation and the technique of SABC immunohistochemistry.Results：30μg/kg polypeptide has obviously inhibiting effect on U251 glioma cells,inhibiting rate of 60 μg/kg reached 53.37%.The positive cells number of PCNA the treatment group is significantly less than the control group.Conclusion：Phlypeptide from agkistrodon acutus venom could inhibit the ugrowth of U251 glioma cells,its pathogenesis can be related with inducing cells apoptosis and inhibiting cell proliferation,which established important theoretical basis for its clinical application.