乌梅丸对乳腺癌小鼠肺转移微环境免疫抑制性细胞因子的影响

Effect of Wumei Pill on immunosuppressive cytokines in lung metastasis microenvironment of breast cancer mice

目的 观察乌梅丸对乳腺癌小鼠肺转移微环境免疫抑制性细胞因子表达的影响。方法 选择8只Balb/c雌性小鼠作为正常组;另选择16只Balb/c雌性小鼠,采用4T1乳腺癌细胞于小鼠第2对乳腺脂肪垫内注射方法构建乳腺癌模型,将造模成功的16只小鼠随机分为乌梅丸组和模型组,每组8只。乌梅丸组给予乌梅丸16.44 g/kg灌胃,正常组和模型组给予等量灭菌饮用水灌胃,均干预1个月。干预完成后处死各组小鼠,分离肺组织,采用免疫荧光组织化学法观察肺组织中环氧化酶2(COX-2)、白细胞介素-6(IL-6)、转化生长因子-β(TGF-β)和血管内皮生长因子(VEGF)表达情况,分别采用RT-qPCR和Western blot法检测肺组织中COX-2、IL-6、TGF-β、VEGF mRNA和蛋白表达情况。结果 模型组肺组织中COX-2、IL-6、TGF-β、VEGF呈强阳性表达,乌梅丸组COX-2、IL-6、TGF-β、VEGF的荧光强度低于模型组。模型组肺组织中COX-2、IL-6、TGF-β、VEGF mRNA和蛋白相对表达量均明显高于正常组(P均<0.05),乌梅丸组COX-2、IL-6、TGF-β、VEGF mRNA和蛋白相对表达量均明显低于模型组(P均<0.05)。结论 乌梅丸可能通过下调免疫抑制性细胞因子COX-2、IL-6、TGF-β、VEGF的表达而抑制乳腺癌肺转移。

Objective It is to observe the effect of Wumei Pill on the expression of immunosuppressive cytokines in the lung metastasis microenvironment of breast cancer mice.Methods Eight Balb/c female mice were selected as the normal group;another 16 Balb/c female mice were selected and injected with 4T1 breast cancer cells into the fat pad of the second pair of mammary glands to establish breast cancer models,and the 16 successfully modeled mice were randomly divided into Wumei Pill group and model group,with 8 mice in each group.The Wumei Pill group was given Wumei Pill 16.44 g/kg by gavage,and the normal group and model group were given equal amounts of sterilized cited water by gavage,and all the groups were intervened for 1 month.After the intervention was completed,the mice in each group were executed,their lung tissues were isolated,and the expression of cyclooxygenase 2(COX-2),interleukin-6(IL-6),transforming growth factor-β(TGF-β)and vascular endothelial growth factor(VEGF)in lung tissues were observed by immunofluorescence histochemistry,and the expression of mRNA and protein of COX-2,IL-6,TGF-βand VEGF in lung tissues were detected by RT-qPCR and Western blot,respectively.Results The expression of COX-2,IL-6,TGF-βand VEGF were strong positive in the lung tissue of the model group,and the fluorescence intensity of COX-2,IL-6,TGF-βand VEGF in the Wumei Pill group was lower than that in the model group.The relative expressions of COX-2,IL-6,TGF-β,VEGF mRNA and protein in lung tissues of the model group were significantly higher than those of the normal group(all P<0.05),while the relative expressions of COX-2,IL-6,TGF-β,VEGF mRNA and protein in the lung tissues of Wumei Pill group were significantly lower than those of the model group(all P<0.05).Conclusion Wumei Pill can inhibit lung metastasis of breast cancer possibly by down-regulating the expression of immunosuppressive cytokines COX-2,IL-6,TGF-βand VEGF.