乳宁冲剂及其拆方对裸鼠移植瘤肺转移的抑制作用与机制探讨

Inhibitory Effect and Mechanism of Runing Granule Preparation and Its Modyfied Prescription on Pulmonary Metastasis of Transplanted Tumou in Naked Mouse

关键词:  乳宁冲剂  乳腺癌  肺转移  实验

Key words:  Runing Granule preparatiion  breast cancer  pulmonary metastasis  in vivo experimental study

摘要:

[目的]通过体内实验探讨乳宁冲剂及其拆方对裸鼠乳腺癌肺转移的抑制作用及其机制。[方法]选雌性BALB/c裸鼠,以对数生长的体外培养的MDA-MB-435高转移乳腺癌细胞造模。随机分为模型组、乳宁冲剂组,乳宁冲剂拆方温肾组、乳宁冲剂拆方疏肝活血组、三苯氧胺组(TAM组)、环磷酰胺组(CTX组),给药7周后,解剖显微镜计数肺转移结节并行全肺组织切片,免疫组化检测肿瘤组织VEGF、MMP-9、TIMP-1表达情况。[结果]乳宁冲剂组及拆方温肾组、疏肝活血组对裸鼠乳腺癌移植瘤的肺转移抑制作用分别为46.36%、32.73%、18.2%,乳宁冲剂组作用与CTX组(50.91%)比较无显著差异;而乳宁冲剂组、疏肝活血组均可下调肿瘤VEGF蛋白表达;乳宁冲剂全方、温肾方可明显使TIMP-1mRNA的表达升高,MMP-9表达下降;对MMP-9/TIMP-1蛋白表达比值影响以温肾组下降最明显。[结论]乳宁冲剂全方抑瘤抗转移作用优于拆方,而拆方组以温肾组抗转移作用优于疏肝活血组。同时表明,乳宁冲剂特别是温肾中药可能通过降低MMP-9/TIMP-1比值而影响肺转移的发生。

Abstract:

[Objective] To discuss the inhibitory effect and mechanism of Runing granule preparation and its modified prescription on pulmonary metastasis of transplanted tumor by in vivo experiment study.[Methods] BALB/c female naked mice were used.The animal models were established by transplanting highly metastatic MDA-MB-435 breast tumor cell.The mice were divided randomly into model group,Runing granule preparation group,warming kidney group,dispersing liver and promoting blood circulation group,tamoxifen(TAM) group and cyclophosphamide(CTX) group.Seven weeks after medication,the number of pulmonary metastasis was accounted by means of anatomic microscope.Slice of lung tissue was obtained to detect the express of VEGF,MMP-9 and TIMP-1 by Immunohistochemistry.[Result] The inhibitory effect on pulmonary metastasis of transplanted tumor were 46.36%,32.73%,18.2% in the Runing granule preparation group,warming kidney group,dispersing liver and promoting blood circulation group respectively.There wasn’t significant difference between Runing granule preparation group and CTX group(50.91%).The expression of VEGF was dropped in Runing granule preparation group and dispersing liver and promoting blood circulation group.The expression of TIMP-1mRNA was up-regulated in Runing Granule group and warming kidney group.The expression of MMP-9 was dropped in Runing granule preparation group and warming kidney group.The ratio of MMP-9 and TIMP-1 expression was obviously lower in warming kidney group.[Conclusion] The tumor-inhibitory effect of Runing granule preparation is better than its modified prescriptioin.The warming kidney group was better than dispersing liver and promoting blood circulation group in the inhibitory effect of metastasis effect.The Runing granule preparation,especially warming kidney group,may affect the occurrence of pulmonary metastasis through decreasing the ratio of MMP-9/TIMP-1.