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[摘要]
目的 阿诺宁(Anuoning)是具有高抗癌活性的番荔枝内酯有效部位药物。对阿诺宁(A)及其主要成分(B)、非主要成分(C)3个组分进行抗肿瘤活性和急性毒性对比试验。方法 用MTT法测定阿诺宁各组分对各种人癌细胞、正常细胞和耐药性(MDR)细胞的细胞毒作用,用小鼠肿瘤S180模型进行体内抗肿瘤试验。应用Bliss方法计算小鼠的半数致死量(LD50)。结果 A、B、C在1×10-4~10.0μg/mL质量浓度时,对人癌细胞CNE2的IC50分别为3.1×10-4、4.3×10-3、0.19μg/mL;对Bel-7402细胞的IC50分别为0.078、0.043、0.39μg/mL;对KB细胞的IC50依次为0.46、0.24和1.26μg/mL;对MCF-7细胞的IC50分别为1.72、2.97、>10μg/mL;在上述质量浓度下对人体正常脐血管内皮细胞(ECV304)的IC50分别为0.39、0.27、0.99μg/mL。在10μg/mL质量浓度时,A、B、C组分对人体正常肝细胞(CLC)的平均生长抑制率均小于20%。A、B、C组分对多药耐药(MDR)细胞MCF-7/Adr的IC50分别为0.32、3.15、>10μg/mL,对KBV200细胞的IC50分别为0.027、0.72、0.83μg/mL。在体内试验中,A在30μg/kg,对小鼠肿瘤S180的3批实验的平均抑瘤率为(52.8±9.5)%(P<0.05~0.001);B在15、30和60μg/kg剂量时的平均抑瘤率依次为(40.1±5.7)%、(50.7±10.3)%、(61.9±3.7)%(P<0.05~0.001);C在45、90和180μg/kg剂量下的平均抑瘤率分别为(32.7±3.4)%、(33.5±0.6)%、(30.6±7.3)%(P<0.05~0.01)。对小鼠ip一次A、B、C的LD50及其95%可信限分别为1.38mg/kg和0.97~1.95mg/kg,0.39mg/kg和0.36~0.43mg/kg,2.39mg/kg和2.12~2.69mg/kg。结论 A、B、C组分在体内外试验均显示不同程度的抗癌活性,其中A的抗肿瘤活性强,毒性较小。MDR细胞对A、B、C组分均不产生抗药性。
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[Abstract]
Objective To compare the antitumor activities and acute toxicity exhibited by Anuoning(A)which is a multi-component drug with potent anti-tumor activity,as well as its two different parts(B)and(C).Methods The cytotoxicities of Anuoning and its two fractions in various human cancers,normal cell,and multidrug resistance(MDR)cell lines were assayed by MTT method.The models of mice sarcoma S180 were used for in vivo antitumor test.Bliss method was used to calculate the lethal dose of 50%(LD50)in mice.Results At the concentrations of(1×10-4-10.0)μg/mL,the IC50 of A,B,and C for human cancer cells(CNE2)were 3.1×10-4,4.3×10-3,and 0.19 μg/mL,respectively;The IC50 for Bel-7402 cells were 0.078,0.043,and 0.39 μg/mL,respectively;The IC50 for KB cells were 0.46,0.24,and 1.26 μg/mL;The IC50 for MCF-7 cells were 1.72,2.97,and 10 μg/mL,respectively.At the concentrations of(1×10-4-10.0)μg/mL,the IC50 of A,B,and C for human normal endothelial cells in navel blood vessel(ECV304)were 0.39,0.27,and 0.99 μg/mL,respectively.At the concentration of 10.0 μg/mL,the average growth inhibitory rates of A,B,and C for human normal liver cells(Chang Liver cell,CLC)were 20%.The IC50 of A,B,and C for multi-drug resistant(MDR)cell lines MCF-7/Adr were 0.32,3.15,and 10 μg/mL and KVB200 cells were 0.027,0.72,and 0.83 μg/mL,respectively.Under the doses of 30 μg/kg,in vivo,the average tumor inhibitory rate of A to mice tumor S180 was(52.8±9.5)%(P<0.05~0.001);Under the doses of 15,30,and 60 μg/kg,the average tumor inhibitory rates of B were(40.1±5.7)%,(50.7±10.3)%,and(61.9±3.7)%(P<0.05~0.001),respectively;Under the doses of 45,90,and 180 μg/kg,the average tumor inhibitory rates of C were(32.7±3.4)%,(33.5±0.6)%,and(30.6±7.3)%(P<0.05-0.01),respectively.In single ip to mice,the LD50 and 95% confidence limit of A,B,and C were 1.38 mg/kg and 0.97-1.95 mg/kg,0.39 mg/kg and 0.36-0.43 mg/kg,and 2.39 mg/kg and 2.12-2.69 mg/kg,respectively.Conclusion Anuoning and its two fractions have obvious antitumor activities in vitro and in vivo.The antitumor activity of A is the highest while the toxicity to mice is lower among them.MDR cell lines have no cross-resistance to A,B,and C.
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[基金项目]
国家自然科学基金资助项目(30572253)