人参皂苷Rg3、索拉非尼和奥沙利铂不同联合方式抗裸鼠肝癌移植瘤血管生成的作用研究

目的 观察人参皂苷Rg3、索拉非尼、奥沙利铂单药及联合对裸鼠人肝癌细胞移植瘤血管生成的作用并探讨其可能机制。 方法 以裸鼠人肝癌转移模型LCI-D20为观察对象，将60只荷瘤裸鼠随机分成8组，分别为人参皂苷Rg3组（A组）、奥沙利铂组（B组）、索拉非尼组（C组）、人参皂苷Rg3+索拉非尼组（D组）、人参皂苷Rg3+奥沙利铂组（E组）、奥沙利铂+索拉非尼组（F组）、人参皂苷Rg3+奥沙利铂+索拉非尼组（G组）和生理盐水对照组（H组）。肿瘤接种第11 d后开始给药，人参皂苷Rg3 5 mg／kg（1天1次）、奥沙利铂5 mg／kg（2天1次），均用腹腔注射，索拉非尼30 mg／kg（1天1次）灌胃。给药14 d后处死裸鼠，剥离瘤体，检测各组瘤组织中微血管密度（MVD），免疫组化染色和Western blotting检测各组瘤组织中血管内皮生长因子（VEGF）、缺氧诱导因子（HIF）-1α和VEGF受体（VEGFR）-2的表达。 结果 与H组比较，其余各组MVD均较低（P＜0.01），VEGF蛋白阳性表达率均较低（P＜0.05），HIF-1α蛋白阳性表达率均下降，但差异无统计学意义（P＞0.05）；A组、D组、E组的VEGFR-2蛋白阳性表达率显著低于H组（P＜0.05）。与H组比较，仅A组、B组、C组、D组、E组能显著下调瘤组织中VEGF、HIF-1α和VEGFR-2蛋白的表达（P＜0.05）。 结论 人参皂苷Rg3具有一定的抗血管生成作用，其机制可能与下调VEGF、HIF-1α、VEGFR-2及MVD的表达有关。人参皂苷Rg3分别与索拉非尼、奥沙利铂联合可以增强抗血管生成作用，但三药联合时抗血管生成作用反而减弱，提示可能存在其他抗肿瘤机制，值得进一步研究。

Experimental study of combination therapy with ginsenoside Rg3, sorafenib and oxaliplatin versus mono-therapy on anti-angiogenic effect in hepatocelluar carcinoma

Objective To observe the effect of ginsenoside Rg3，sorafenib，oxaliplatin alone or with different combination in neovascularization of tumor in nude mice with human hepatocellular carcinoma， and investigate the possible mechanisms. Methods Sixty nude mice with high metastatic human hepatocellular carcinoma transplanted in situ（LCI-D20）were randomly divided into ginsenoside Rg3 group（Group A），oxaliplatin group（Group B），sorafenib group（Group C），ginsenoside Rg3+ sorafenib group（Group D），ginsenoside Rg3+oxaliplatin group（Group E），oxaliplatin+sorafenib group（Group F），ginsenoside Rg3+ oxaliplatin+sorafenib group（Group G）and formal saline group（Group H）. The drug was administered eleven days later after the inoculation of tumor. The dosage of the medicine was as follows：ginsenoside Rg3（5mg／kg， qd， 14 days， intraperitoneal injection） and oxaliplatin（5mg／kg， q2d， 14 days， intraperitoneal injection） and sorafenib（30mg／kg， qd， 14 days， gastric lavage）. All mice were sacrificed at 14 days after administration，and tumor tissues were resected. Immunohistochemical method and Western blotting were used to detect the expression rate and relative quantity of vascular endothelial growth factor（VEGF）， hypoxia-inducible factor（HIF）-1α， VEGF receptor（VEGFR）-2 and micro-vessel density（MVD） in the tumor. Results The treatment groups presented different degrees of MVD lower than that of Group H（P＜0.01）. Positive expression rates of VEGF in all treatment groups were decreased significantly compared with the Group H（P＜0.05）. Compared with Group H， positive expression rates of HIF-1α had a overall downward trend（P＞0.05）.The positive expression rates of VEGFR-2 in Group A， D and E were decreased significantly than that in Group H（P＜0.05）. Relative quantity of VEGF， HIF-1α and VEGFR-2 in Group A， B， C，D and E were decreased significantly than those in Group H（P＜0.05）. Conclusion Ginsenoside Rg3 has an anti-angiogenic effect. Its anti-angiogenic effect might be related to the down-regulated expression of VEGF， HIF-1α and VEGFR-2. Ginsenoside Rg3 in combination with oxaliplatin or sorafenib can enhance the anti-angiogenic effect， but the expression of VEGF， HIF-1α and VEGFR-2 protein didn’t decreased significantly in the combination of three drugs. It shows that the antitumor effect of three drugs may still relate other mechanisms, which needed further study.