醋制法对瑞香狼毒毒效影响的研究

Study on impact of vinegar processing on toxic effect of Stellera chamaejasme

目的:为探讨有毒中药瑞香狼毒炮制解(减)毒理论的科学性,并了解瑞香狼毒醋制前后对药效的影响。方法:PHLC-MS技术比较瑞香狼毒醋制前后成分差异;建立H22皮下移植瘤模型,比较醋制前后对荷瘤小鼠与正常小鼠的致死作用及体重变化;荷瘤小鼠连续灌胃给药后比较醋制前后对肿瘤及免疫器官的影响;荧光报告基因法研究瑞香狼毒提取物对肿瘤细胞作用的靶标基因TGF-β,AP1,NF-κB的影响。结果:瑞香狼毒提取物Zp1102与醋制后的提取物Zp1103的LD50分别为9.89,16.85 g·kg-1,Zp1103半数致死剂量高于Zp1102。药效试验表明,药物为2 g·kg-1时,Zp1102对小鼠皮下移植瘤H22有显著抑制作用,抑瘤率为36.24%(P<0.01),Zp1103的抑瘤率略低,为34.40%(P<0.05);药物为1 g·kg-1时,Zp1102抑瘤率为34.52%(P<0.05),而Zp1103抑瘤率明显降低,为21.55%。Zp1102,Zp1103对报告基因AP1基本无影响;Zp1102对HepG2细胞经刺激内源性NF-κB浓度升高后的报告基因有上调作用,且能下调TGF-β的表达,但Zp1103仅能上调NF-κB的表达,对TGF-β无影响。结论:瑞香狼毒醋制后的提取物较未经炮制而提取工艺相同的提取物毒性降低,同时体内药效学实验结果表明其抗肿瘤活性也略有降低,其机制可能与药物对转化生长因子TGF-β的调节有一定关联。

Objective ： To study the scientific theory on detoxification （attenuation） of Stellera chamaejasme （ScL） by processing and the impact on drug effect of ScL before and after being processed with vinegar. Method： The difference in ingredients of ScL before and after being processed vinegar was compared by using PHLC-MS technique. A subcutaneously transplanted tumor model of H22 hepatoma was established to compare the lethal effect and weight change between tumor-loaded mice and normal mice. After consecutive oral administration in tumor-loaded mice, the impacts on tumors and immune organs were compared before and after being processed with vinegar. Luciferase Report Gene was employed to investigate the target genes TGF-β, AP1 and NF-KB. Result： The LD50 （median fatal dose） of extract Zp1102 exhibited higher than that of the processed one Zp1103, that is 9.89 g·kg^-1 vs 16. 85 g ·kg^-1. According to the test, Zp1102 showed more effective anti-tumor activities in vivo than that of Zp1103 in a same dosage, with the tumor inhibitory rate 36. 24% （P 〈 0. 01 ） at the dosage of 2 g·kg^-1 and 34.40% （P 〈 0. 05 ） at the dosage of 1 g ·kg^-1. At the dosage of 1 g ·kg^-1, Zp1102 showed a tumor inhibitory rate of 34. 52% （P 〈 0. 05 ）, much higher from 21.55% in Zp1103. Both Zp1102 and Zp1103 had basically no impact on the report gene NF-KB, besides that Zp1102 up-regulated the report gene after increase in NF-KB concentration and down-regulated TGF-β, but Zp1103 can only up-regulate NF-KB expression without any impact on TGF-β. Conclusion： Processed ScL extracts show less toxic than unprocessed extracts and slight reduction in anti-tumor activity, which may be related to the regulation of transforming growth factor TGF38.