新加沙参麦冬煎剂抑制肿瘤转移及其作用机制的实验研究

An Experimental Study of the Inhibitory Effect of Tailored Shashen Maidong Decoction on Tumor Metastases and Its Underlying Mechanisms

目的探讨新加沙参麦冬煎剂防治肿瘤转移的机制。方法结合动物模型、免疫组化、细胞分子生物学技术,观察新加沙参麦冬煎剂对荷瘤小鼠抑瘤率、转移抑制率、生命延长率的影响,检测实验小鼠皮下移植瘤血管内等细胞生长因子(VEGF)、血管内皮细胞第Ⅷ因子(CD34)、黏附因子(CD44V6)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶抑制酶(TIMP2)的表达情况。结果新加沙参麦冬煎剂治疗组的瘤重抑制率、肺转移抑制率、生命延长率分别为37.3%,58.3%和20.8%;皮下移植瘤CD44V6、VEGF表达、微血管密度(MVD)明显低于空白组(P<0.01),TIMP2表达记分明显高出其他组(P<0.01)。小鼠肺转移灶数与VEGF、CD44V6、MVD呈线性相关,相关系数分别为0.490,0.398,0.455。结论①新加沙参麦冬煎剂对小鼠LA795高转移肺腺癌模型有较好抑制转移、抑制肿瘤生长和延长荷瘤小鼠生存时间的作用。②新加沙参麦冬煎剂可通过调控肿瘤转移过程中黏附、基质降解、血管生成相关分子的表达,多途径、多靶点防治肿瘤的转移。

Objective To probe into the effects of the tailored shashen maidong decoction（TSMD） in the prevention and treatment of tumor metastases in mice and the underlying mechanisms. Methods Aliquots of the suspension containing tumor cells of the highly metastatic hmg adenocarcinoma LA795 in mice were inoculated subcutaneously into T739 mice. 24h after the challenge, mice bearing the tumor were randomly divided into 3 equal groups （n = 20 in each group ）： the cyclophophamide（Cy） group, the TSMD group and the blank control group. Mice of the Cy group were given each an intraperitoneal injection of 60 mg·kg^-1 of Cy q. o.d. for 16 consecutive days. Mice of the TSMD group were given each 0.45 mL· d^-1 of TSMD q. d. administered by gastrogavage for 16 consecutive days. Mice of the blank control group were given each 0. 45 mL· d^-1 of distalled water q.d. administered by gastrogavage for 16 consecutive days. The rate of tumor inhibition as reflected by the decrease in the weight of the subcutaneous tumor 20 days after the beginning of the drug treatment, the rate of inhibition of lung metastases, the survival span of the animals and expression of V EGF （vascular endothelial growth factor） , CD34 （vascular endothelial factor VS ）, CD44V6 （ adhesion molecule）, MMT2 （ matrix metalloproteinase 2 ） and TIMP2 （ tissue inhibitor of metalloproteinase 2） by the subcutaneous tumors in mice of the different groups were compared. Results The rate of tumor inhibition in mice treated with TSMD was 37.3%, which was , however , lower than that （57.3%）in mice treated with Cy（ P 〈 0.01 ）. The survival span of mice treated with TSMD was （31.4 ±2.5） days , which was significantly longer than that （26.0 ±3. 1 days） in mice of the blank control group but similar to that （ 30.1±3.6 days） in mice treated with Cy （ P 〉 0.05）, The expression of VEGF by the subcutanecous tumor in mice treated with TSMD was （2.6±1.4）, which was significantly lower than that （5.2±3.2） in mice of the blank control group （P 〈 0.01 ） but similar to that （4.7±2.5 in mice treated with Cy（ P 〉 0. 05）. The microvascular density （MVD） of the subcutaneous tumor in mice treated with TSMD was 10.2 ±1.7, which was strikinly lower than that （ 15.7 ± 4.4） in mice of the blank control group （P 〈 0. 01 ） but similar to that （ 10.2 ±1.7） in mice treated with Cy （ P 〉 0.05 ）. The expression of CD44V6 by the subcutaneous tumor in mice treated with TSMD was （2.1±1.9）, which was significantly lower than that （4.1±2.6） in mice of the blank control group （P 〈 0.01 ） and than that （5.0±2.9） in mice treated with Cy （P 〈 0.01 ）. The expression of TIMP2 by the subcutaneous tumor in mice treated with TSMD was （4.7±2.4）, which was significantly higher than that （2.8 ±1.9） in mice of the blank control group （ P 〈 0. 01 ） but similar to that （3.6 ±1.5） in mice treated with Cy（P 〉0.05） . The expression of MMP2 by the subcutaneous tumor in mice of the blank control group and those treated with TSMD and Cy were （4.8 ± 3.6 ）, （4.6 ± 2.4 ） and （ 5. 1± 3.0）, respectively, the differences between them being insignificant（ P 〉 0. 05, P 〉 0. 05 ）. A linear correlation was demonstrated between the number of lung metastases and VEGF, CIM4V6 and MVD, the correlation coefficients being 0. 490,0. 398 and 0. 455, respectively. Conclusion ①TSMD was shown to inhibit tumor growth and lung metactaces, prolong survival span in mice bearing highly metastatic lung adenocarcinoma LA795. ②TSMD is thought by the authors to inhibit tumor metastases in a multi-pathway and multi-target manner by modulating adhesion, matrix degradation and expression of corresponding molecules relevant to angiogenesis during the process of tumor metatases.