十全大补汤抗小鼠黑色素瘤作用及与顺铂联合用药的研究

目的 探讨十全大补汤（SQDB）对小鼠黑色素瘤的抑制作用及机制，阐明其与顺铂（DDP）联合用药的作用效果及给药方式。方法 流式细胞术考察SQDB对小鼠黑色素瘤细胞B16F10细胞周期及凋亡的影响；分离小鼠脾细胞，MTS法考察SQDB对刀豆蛋白A（Con A）及脂多糖（LPS）促进小鼠脾细胞向T、B淋巴细胞转化的影响；构建小鼠黑色素瘤皮下移植瘤模型，通过 HE染色、免疫组化、TUNEL染色等方法考察SQDB的抑瘤作用；构建小鼠黑色素瘤转移瘤模型，探讨SQDB抗肿瘤转移作用及潜在机制。结果 10 mg/mL的SQDB能诱导B16F10黑色素瘤细胞凋亡；2.5、5、10 mg/mL SQDB减少了G₀/G₁期细胞数；10 mg/mL SQDB增加了S期细胞数；2.5 mg/mL SQDB增加了G₂/M期细胞数。5%和10%含药血清能够促进Con A所致B淋巴细胞增殖；5%、10%和15%含药血清对于LPS所致T淋巴细胞增殖亦有一定促进作用。同时，SQDB与DDP联用能够显著抑制肿瘤细胞增殖，促进肿瘤细胞凋亡；在小鼠黑色素瘤转移模型中，DDP与SQDB同时合用，或者先用DDP再用SQDB均能显著抑制黑色素瘤肺转移；两者同时给予能够显著提高小鼠胸腺指数，而先给SQDB再给DDP则显著逆转小鼠脾指数下降的现象，单用SQDB能够降低肿瘤组织局部IL-1β水平，促进IL-4分泌，从而抑制肿瘤转移。结论 SQDB能够抑制黑色素瘤B16F10增殖，促进其凋亡及小鼠脾淋巴细胞增殖；与DDP同时或贯序使用能够增强对小鼠移植瘤及转移瘤模型的抑制作用。 

Study on the Anti-Melanoma Effect of Shiquan Dabu Decoction and its Combination Therapy with Cisplatin

OBJECTIVE To investigate the anti-tumor effect and mechanism of Shiquan Dabu Decoction （SQDB）on mouse melanoma， and to elucidate the effect and administration of the combination with cisplatin（DDP）. METHODS Flow cytometry was used to investigate the effect of SQDB on cell cycle and apoptosis of melanoma cell line B16F10. Mouse spleen cells were isolated and MTS assay was used to investigate the anti-proliferative activity of SQDB in spleen cells induced by ConA or LPS. The model of mouse melanoma subcutaneous xenograft was established. The anti-tumor effect of SQDB was detected by HE staining， immunohistochemistry and TUNEL staining. The melanoma metastasis model was constructed to study the potential mechanism on the inhibition of the tumor metastasis by SQDB. RESULTS 10 mg/mL SQDB could induce apoptosis of B16F10 melanoma cells; 2.5， 5， 10 mg/mL SQDB reduced the number of G₀/G₁ cells; 10 mg/mL SQDB increased the number of S phase cells; 2.5 mg/mL of SQDB increased the number of cells in G₂/M phase; 5% and 10% drug-containing serum promoted the proliferation of B lymphocytes induced by LPS; 5%， 10% and 15% drug-containing serum also promoted the proliferation of T lymphocytes caused by Con A. Using DDP and SQDB simultaneously or using DDP first and then SQDB inhibited the growth of melanoma. Using both DDP and SQDB significantly inhibited tumor cell proliferation and promoted tumor cell apoptosis; When DDP and SQDB were used together， or when DDP was used first， SQDB could significantly inhibit melanoma lung metastasis. In addition， the thymus and spleen coefficient of the mice decreased significantly when DDP alone was used (P<0.05， P<0.01)， and the simultaneous administration of SQDB and DDP could increase the thymus coefficient of the mice (P<0.05). CONCLUSION SQDB can inhibit the proliferation of melanoma B16F10， promote B16F10 apoptosis and the proliferation of mouse spleen lymphocytes. Combined with DDP， SQDB can enhance the anti-tumor effect of mouse xenografts and metastatic tumor models.