益肺清化颗粒对Lewis肺癌小鼠VEGF、bFGF、Angiostatin、Endostatin影响的研究

Effect of Yifei Qinghua Granule on VEGF, bFGF, Angiostatin, and Endostatin in Lewis Lung Cancer Mice: an Experimental Study

目的观察中药益肺清化颗粒对Lewis肺癌小鼠瘤组织中的血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、血管抑素(Angiostatin)、内皮抑素(Endostatin)的影响,探讨益肺清化颗粒的抑瘤机制。方法 70只小鼠随机分为模型组,中药低、中、高剂量组,吉非替尼组,吉非替尼加中药中剂量组,以及环磷酰胺(CTX)组,每组10只。所有小鼠右腋皮下接种Lewis瘤细胞进行造模。模型组给予纯净水0.4mL灌胃,每天1次;中药低、中剂量组分别给予益肺清化颗粒5、10g/kg灌胃,每天1次;中药高剂量组给予益肺清化颗粒10g/kg灌胃,每天2次;吉非替尼组给予吉非替尼100mg/kg灌胃,每天1次;吉非替尼加中药中剂量组上午给予吉非替尼100mg/kg灌胃,下午给予益肺清化颗粒10g/kg灌胃。上述各组均从小鼠接种肿瘤细胞的第2天开始给药,连续14天。CTX组在实验第3、7天给予CTX60mg/kg腹腔注射。第15天处死小鼠,取瘤,免疫组化法检测肿瘤组织中VEGF、bFGF、Angiostatin、Endostatin的表达。结果与模型组比较,各给药组VEGF表达均显著降低(P<0.01),Angiostatin、Endostatin表达显著增加(P<0.01),且吉非替尼组bFGF表达明显降低(P<0.05)。各给药组间VEGF比较,差异无统计学意义(P>0.05)。CTX组Angiostatin表达显著高于中药低剂量组(P<0.01)。中药高剂量组、吉非替尼加中药中剂量组Endostatin表达明显高于中药低、中剂量组(P<0.01);吉非替尼加中药中剂量组Endostatin表达明显高于吉非替尼组(P<0.05)。结论益肺清化颗粒可能通过降低血管生成促进因子VEGF的表达,升高血管生成抑制因子Angiostatin、Endostatin的表达,进而发挥抑瘤作用。

Objective To observe the effect of Yifei Qinghua Granule （YQG~） on vascular endo- thelial growth factor （VEGF）, basic fibroblast growth factor （bFGF）, angiostatin, and endostatin in tumor tissue of Lewis Lung cancer mice, and to explore its anti-tumor mechanisms. Methods Totally 70 C57BL/6 mice were randomly divided into the model group, the low, medium, and high dose YQG groups, the gefitinib group, the gefitinib plus medium dose YQG group, and the cyclophosphamide （CTX） group, 10 in each group. The models were established by subcutaneously injecting Lewis lung cancer cells from the right axilla of C57BL/6 mice. Mice in the model group were given with 0.4 mL pure water by gastrogavage, once daily. Mice in the low and medium dose YHG groups were given with YHG at the daily dose of 5 and 10 g/kg by gastrogavage, once daily. Those in the high dose YHG group were given with YHG at 10 g/kg by gastrogavage, twice daily. Those in the gefitinib group were given with gefitinib 100 mg/ kg by gastrogavage, once daily. Those in the gefitinib plus medium dose YHG group were given with ge- fitinib at 100 mg/kg by gastrogavage in the morning and YHG at 10 g/kg by gastrogavage in the afternoon. All medication was started from the 2nd day of innoculation, lasting 14 successive days. Those in the CTX group were given CTX at 60 mg/kg by peritoneal injection on the 3rd and the 7th day of the experi- ment. Mice were sacrificed at the fifteenth day of the experiment. Tumors were taken out. Expressions of VEGF, bFGF, angiostatin, and endostatin in the tumor tissue were detected using immunohistochemical assay. Results Compared with the model group, the expression of VEGF significantly decreased, ex- pressions of angiostatin and endostatin significantly increased in each group （P 〈0.01 ）. The expression of bFGF significantly decreased in the gefitinib group （P 〈0.05~. There was no statistical difference in VEGF among all groups （P 〉0.05）. The angiostatin expression was significantly higher in the CTX group than in the low dose YQG group （P 〈0.01 ）. The expression of endostatin was significantly higher in the high dose YQG group and the gefitinib plus medium dose YQG group than in the low and the medium dose YQG groups （P 〈0.01 ）. The expression of endostatin was significantly higher in the gefitinib plus medium dose YQG group than in the gefitinib group （P 〈0.05）. Conclusion The action mechanism of YQG in treating lung cancer might be achieved through reducing the expression of angiogenesis promoting factor VEGF and increasing expressions of angiogenesis inhibitors angiostatin and endostatin.