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目的探讨中药肺积方抑制裸鼠移植瘤组织血管生成及促进肿瘤血管正常化的作用及其机制。方法采用人肺腺癌细胞株SPAC-1建立BALB/c裸鼠鼠背视窗模型,造模后将裸鼠随机分为空白组及肺积方高、中、低剂量组。用共聚焦显微镜活体观察各组裸鼠活体背部视窗的肿瘤内部血管,计算瘤内单位面积的平均血管数量和直径;同时用免疫荧光染色法观察肿瘤血管壁Ⅳ型胶原(Collagen-Ⅳ)及血管平滑肌抗体(a-SMA)细胞覆盖率。结果在干预后第5天,高剂量组小鼠瘤径显著小于空白组(P<0.05);第9天,高剂量组与中剂量组血管数明显少于空白组(P<0.05),且高剂量组的血管平均直径明显小于中剂量组(P<0.05);第14天,高剂量组、中剂量组血管数明显少于空白组(P<0.05),且高剂量组瘤径明显小于空白组和低剂量组(P<0.05)。自药物干预第6天始,肿瘤血管a-SMA和Collegen-Ⅳ覆盖率较空白组逐步升高,其中高、中剂量组较空白组升高明显(P<0.05)。结论肺积方具有抑制肿瘤血管生成并可促进肿瘤血管正常化的作用,其机制与增加肿瘤血管aSMA和Collegen-Ⅳ覆盖率相关。
Objective To investigate the effect and mechanism of Chinese medicine Jifang Fang on angiogenesis and tumor vascular normalization in xenografted tumor of nude mice. Methods The model of posterior window of BALB / c nude mice was established by human lung adenocarcinoma cell line SPAC-1. The nude mice were randomly divided into blank group and high, medium and low dose groups. Confocal microscopy was used to observe the internal blood vessels of tumor in vivo and in vivo. The average number and diameter of blood vessels per unit area were calculated. Immunofluorescence staining was used to observe the expression of Collagen-Ⅳ and vascular Smooth muscle antibody (a-SMA) cell coverage. Results On the fifth day after intervention, the tumor diameter of the high dose group was significantly smaller than that of the blank group (P <0.05). On the ninth day, the number of vessels in the high dose group and middle dose group was significantly less than that in the blank group (P <0.05) (P <0.05). On day 14, the numbers of blood vessels in high-dose group and medium-dose group were significantly less than those in blank group (P <0.05), and the diameter of high-dose group was significantly smaller than that of middle-dose group The blank group and low dose group (P <0.05). The coverage of a-SMA and Collegen-Ⅳ in tumor vessels gradually increased from the sixth day after drug intervention compared with the blank control group, and the levels of tumor cells in the high and middle dose groups increased significantly compared with the blank control group (P <0.05). Conclusion Feiji Fang can inhibit the tumor angiogenesis and promote the normalization of tumor vessels. The mechanism is related to increasing the coverage of tumor aSMA and Collegen-Ⅳ.