舟山眼镜蛇毒抗肿瘤有效组分分离及其抑瘤作用研究初探

Experimental studies on tumor-inhibiting effects of fractionated components from snake venom of Naja atra Cantor

目的从舟山眼镜蛇(Naja atraCantor)蛇毒(snake venom,SV)中分离得蛇毒组分,探讨SV及其分离组分的LD50和抑制肿瘤的作用。方法采用凝胶柱层析方法从蛇毒中分离得到了前Ⅰ1、Ⅰ1、Ⅱ1、Ⅱ2、Ⅲ1、Ⅲ2及Ⅳ等7种组分。采用急性毒性实验、MTT法,研究SV及其7种SV分离组分的LD50和抑制肿瘤的作用。结果 SV经Sephadex G-50层析,可分离为前Ⅰ、Ⅰ、Ⅱ、Ⅲ及Ⅳ5个组分,根据峰面积大小排列:Ⅲ>Ⅱ>Ⅰ>Ⅳ>前Ⅰ。5个组分再经Sephadex G-25柱层析,可获得7个脱盐组分:前Ⅰ1、Ⅰ1、Ⅱ1、Ⅱ2、Ⅲ1、Ⅲ2及Ⅳ。通过急性毒性实验,明确Ⅳ的毒性最大,其次为Ⅲ2及Ⅲ1,三者的LD50值均低于SV;而Ⅰ1、Ⅱ1、Ⅱ2的毒性均小于SV,前Ⅰ1几乎无毒。SV组分Ⅲ2和Ⅳ的抑瘤作用最强,在高浓度(20μg/ml)时对实验中的2种人肿瘤细胞的抑制率均达到60%以上。结论从SV中分离得到了前Ⅰ1、Ⅰ1、Ⅱ1、Ⅱ2、Ⅲ1、Ⅲ2以及Ⅳ7种组分;组分Ⅳ毒性最强,依次为Ⅲ2>Ⅲ1>SV>Ⅱ2>Ⅱ1>Ⅰ1>前Ⅰ1;SV及其7种分离组分对2种人肿瘤细胞株(SGC-7901、A549)的生长抑制有一定的特异性,而不同的SV分离组分对同一肿瘤细胞抑制作用亦有差异。

Objective Some fractions were isolated from snake venom of Naja atra Cantor by using gel column chromatography.Studies on tumor-inhibiting effects of fractionated components from snake venom.Methods Acute toxicity of these fractions was detected by using mouse acute toxicity test.The antitumor effects of snake venom fractions were further observed in this experiment.By using MTT test.Results Five fractions,including Front Ⅰ1,Ⅰ1,Ⅱ,Ⅲ and Ⅳ were isolated from snake venom of Naja atra Cantor by using Sephadex G-50 gel column chromatography.Two fractions were isolated from Ⅱ or Ⅲ respectively by using Sephadex G-25 gel column chromatography for desalinization.According to the values of LD50,the toxicity of Ⅳ is maximum in seven initial fractions,and then following by Ⅲ2Ⅲ1SVⅡ2Ⅱ1Ⅰ1FrontⅠ1.Snake venom fractions Ⅲ2 and Initial Ⅳ possess strongest tumor inhibiting effect,inhibiting rate of 6 kinds of human tumor cells was more than 60% when their concentration was 20 μg/ml.Conclusion Seven fractions have isolated from snake venom of Naja atra Cantor,including Front Ⅰ1,Ⅰ1,Ⅱ1,Ⅱ2,Ⅲ1,Ⅲ2 and Ⅳ.The toxicity of Ⅳ is maximum and then following by Ⅲ2Ⅲ1SVⅡ2Ⅱ1Ⅰ1FrontⅠ1.Seven kinds of snake venom fractions can inhibit the two kinds of human tumor cells growth（SGC-7901,A549）.Tumor-inhibiting effect of snake venom fractions is different.