健脾理气中药对荷B16黑色素瘤小鼠肝转移的干预作用

Intervention of Spleen-Nourishing and Qi-Enriching Herbs on Liver Metastasis of B16 Melanoma Grafted in Mice

目的观察健脾理气中药对C57BL/6小鼠肝转移的干预作用。方法雌性小鼠50只,随机分为空白组、荷瘤组、脾虚荷瘤组、中药荷瘤组、中药脾虚荷瘤组,每组各10只。以异位接种法在C57BL/6小鼠中建立B16黑色素瘤肝转移模型。空白组、荷瘤组予灌胃冷开水,脾虚荷瘤组灌胃大黄合剂,中药荷瘤组灌胃健脾理气合剂,中药脾虚荷瘤组予灌胃大黄合剂与健脾理气合剂。连续给药12d,停药后次日将所有动物脱椎处死,眼球摘除法取血进行血浆分离;对血清DNA进行定量并观察健脾理气中药对荷瘤肝转移小鼠生存质量、肿瘤生长和肝转移的影响。结果在正常小鼠中,血循环DNA为(62.4±25.52)ng/ml、荷瘤伴有肝转移小鼠为(241±73.7)ng/ml(P<0.01);用健脾理气中药抑制肿瘤生长和转移的同时,荷瘤小鼠血循环DNA也同步下降。结论健脾理气中药可显著地抑制小鼠B16黑色素瘤脾脏种植瘤的生长,能降低脾虚荷瘤组肝转移的发生率,降低脾虚荷瘤组肝转移的分期,降低荷瘤小鼠血微量DNA。

Objective To ascertain the spleen-nourishing and qi-enriching herbs＇ effects on the tumor and the liver metastasis in experimental mice grafted with B16 melanoma. Methods Fifty female mice were divided randomly into blank group, tumor group, insufficiency of spleen plus tumor group, Chinese herbs plus tumor group and Chinese herbs, insufficiency of spleen plus tumor group, 10 mice in each group. Liver metastasis model was replicated after duplicating spleen-deficiency model in C57BL/6 mice. Stomach-perfusion with cold boiled water was given in blank and tumor groups, Rhubarb Mixture in insufficiency of spleen plus tumor group, ＂Jianpi Liqi Mixture＂ in Chinese herbs and tumor group plus Rhubarb Mixture, and ＂Jianpi Liqi Mixture＂ in Chinese herbs, insufficiency of spleen plus tumor group. The medicine was given for successive 12 days. On the next day of withdrawing medicine, all the mice were sacrificed. The blood was taken by removing eyeball and blood plasma was separated. The effects of spleen-nourishing and qi-enriching herbs on the inhibition rate of spleen tumors and the liver metastasis were observed respectively. Serum DNA was extracted by commercial ＂genomic DNA extraction kit＂. Results The mean value of circulating DNA in the healthy group was （62.4±25.5）ng/ml, and that in the mice with liver metastasis group was （241± 73.7） ng/ml, there was a significant difference between the two groups （P〈0.01）. Spleen-nourishing and qi-enriching herbs could suppress the tumor growth and its liver metastasis, down-regulate the mice with liver metastasis in the ＂Jianpi Liqi Mixture＂ plus insufficiency of spleen group, depress the amount of circulating plasma DNA. Conclusion Circulating DNA can reflect the degree of mice with neoplasm and the effect of the drug. Spleen-nourishing and qi-enriching herbs can reduce the value of circulating plasma DNA and improve the efficacy on the liver metastasis.