中华眼镜蛇毒组分对神经胶质瘤细胞U251生长的抑制作用

The Growth Inhibitory Effects of Snake Venom from Naja Naja Actra on Human Glioma U251 Cell Line

目的：探讨中华眼镜蛇毒卡迪（KD）中进一步分离纯化的10个组分KDⅠ-1、KDⅠ-1H、KDⅠ-2、KDQⅠ-2、KDⅡ、KDⅡ-2、KDⅡ-3，KDⅢ-1，KDⅢ-2和KDⅢ-2对神经胶质瘤细胞U251的抑制作用。方法：采用MTT法和台盼蓝拒染法观察各组分对U251细胞增殖的影响。结果：在1～100μg/ml．的浓度范围内用MTT法筛选出3个具有显著抑制作用的组分KD、KDⅡ-3和KDⅢ-1，对以上3种组分再次细分浓度后（5、10、30、50、80、100μg/mL）的MTT结果显示以上三种组分仍然具有显著的抑制作用。选取了KDⅢ-1和KDⅡ-3进一步细分浓度后（1、3．75、7．5、15、30μg/ml，）的MTT结果显示KDⅢ-1和KDⅡ-3组分在7．5μg/mL时的抑制率分别为63．94％和62．15％，且2种组分在1～30μg/mL。呈现剂量效应关系（r=0．694，P〈0．05；r=0．732，P〈0.05）。半数抑制浓度分别为6．65μg/mL；和10。54μg/mL，KDⅡ-3的生长曲线显示药物的各个浓度（1、3．75、7．5、15、30μg/mL）对肿瘤细胞均有抑制作用，以24h的抑制作用最为明显。在7．5-15μg/mL浓度范围内呈现明显抑制作用。结论：分离纯化的10组分对U251细胞有不同程度的抑制作用，无以KDⅡ-3的抑制作用最为显著，并有明显的剂量效应关系。

Objective：To observe the growth inhibitory effects of 10 peplides （KD Ⅰ-1, KD Ⅰ-1 H, KD Ⅰ-2, KD Q Ⅰ-2, KD Ⅱ, KD Ⅱ-2, KD Ⅱ-3, KDⅢ-1 , KD Ⅲ-2, KD Ⅲ-2＇） purified from KD, one of components in Naja Naja Actra venom, on human glioma U251 cell line. Methods： The growth inhibition by each peptide in U251 was studied with MTT assay and trypan blue exclusion. Results： MTT assay screened out three eomponents （KD, KD Ⅱ-3 and KD Ⅲ-1 ） with significant growth inhibition on U251 within 1-100μg/mL interval of levels. MTr assay of these components at further classified levels （5, 10, 30, 50, 80 and 100μg/mL） reconfirmed such inhibitory effects. When KD Ⅱ-3 and KD Ⅲ-1 were selected to be tested at more specified sublevels （ 1, 3.75, 7.5, 15 and 30μg/mL） , maximum rates of inhibition were observed for KD Ⅲ-1 （63.94%） and KD Ⅱ-3 （62.15%） at a level of 7.5ug/mL. Furthermore, the both showed dose-effect correlations （r = 0. 694, P 〈 0. 05 ; r = 0. 732, P 〈 0.05, respectively） with 1-30ug/mL interval of levels. The IC50 was 6.65ug/mL and 10.54ug/mL, respectively. The KDⅡ-3 inhibited growth curve suggested evidenced inhibition on U251 at all specified sublevels, which appeared to be more remarkable with the 7.5-15μg/mL interval, and was most significant at 24 hours. Conclusion： 10 KD-purified peptides were shown to have varied inhibitory effect on the growth of U251. Among them, KDⅡ-3 resulted in the most remarkable inhibition with an identifiahle dose-effect correlation.