益气通窍方对鼻咽癌裸鼠的抑瘤作用及机制

Anti-Tumor Effect and Mechanism of Yiqi Tongqiao Prescription onNasopharyngeal Carcinoma in Nude Mice

【目的】探讨益气通窍方对鼻咽癌裸鼠的治疗作用及机制。【方法】将32只裸鼠随机分为正常组,模型组及益气通窍方低、高剂量组,每组8只。除正常组外,其余各组裸鼠构建鼻咽癌模型。治疗结束后,测量裸鼠瘤体体积,采用酶联免疫吸附分析(ELISA)检测血清转化生长因子β1(TGF-β1)、血管内皮细胞生长因子(VEGF)水平,苏木素-伊红(HE)染色法观察鼻黏膜组织病理形态,定量聚合酶链反应(PCR)法检测鼻黏膜组织基质金属蛋白酶组织抑制剂1(TIMP-1)、基质金属蛋白酶9(MMP-9)的mRNA表达,蛋白免疫印迹(Western Blot)法检测鼻黏膜组织Wnt-1、β-连环蛋白(β-catenin)蛋白表达。【结果】与正常组比较,模型组裸鼠瘤体体积增大,血清TGF-β1、VEGF水平升高,鼻黏膜组织TIMP-1 mRNA表达水平降低、MMP-9 mRNA表达水平升高,鼻黏膜组织Wnt-1、β-catenin蛋白表达水平升高(均P<0.05),HE染色可见鼻黏膜结构缺损,大量嗜酸性粒细胞及其他炎性细胞浸润;与模型组比较,益气通窍方低、高剂量组裸鼠瘤体体积缩小,血清TGF-β1、VEGF水平降低,鼻黏膜组织TIMP-1 mRNA表达水平升高、MMP-9 mRNA表达水平降低,鼻黏膜组织Wnt-1、β-catenin蛋白表达水平降低(均P<0.05),呈剂量依赖性,HE染色可见鼻黏膜细胞形态明显改善。【结论】益气通窍方通过TIMP-1的内源性调节作用抑制MMP-9活性,维持鼻咽癌细胞外基质稳态,通过抑制Wnt/β-catenin信号通路的异常激活,减少TGF-β1、VEGF释放,减少肿瘤血管生成,进而实现对鼻咽癌裸鼠的抑瘤作用。

Objective To investigate the therapeutic effect and mechanism of Yiqi Tongqiao Prescription on nasopharyngeal carcinoma in nude mice.Methods Thirty-two nude mice were randomly divided into normal group,model group,Yiqi Tongqiao Prescription low-and high-dose groups,with eight mice in each group.Except for the normal group,the other groups of nude mice were used to construct a nasopharyngeal carcinoma model.After treatment,the tumor volume of nude mice was measured,the levels of serum transforming growth factorβ1(TGF-β1)and vascular endothelial growth factor(VEGF)were detected by enzyme-linked immunosorbent assay(ELISA),the pathological morphology of nasal mucosa was observed by hematoxylin-eosin(HE)staining,the mRNA expressions of tissue inhibitor of metalloproteinase 1(TIMP-1)and matrix metalloproteinase 9(MMP-9)in nasal mucosa were detected by quantitative polymerase chain reaction(PCR),the protein expressions of Wnt-1 andβ-catenin in nasal mucosa were detected by Western Blot.Results Compared with the normal group,the tumor volume of nude mice in the model group was increased,the levels of serum TGF-β1 and VEGF were increased,the mRNA expression level of TIMP-1 in nasal mucosa was decreased,the mRNA expression level of MMP-9 was increased,and the protein expression levels of Wnt-1 andβ-catenin in nasal mucosa were increased(all P<0.05).HE staining results showed structural defects of nasal mucosa,and a large number of eosinophils and other inflammatory cell infiltration.Compared with the model group,the tumor volume of nude mice in the low-and high-dose groups of Yiqi Tongqiao Prescription was reduced,the levels of serum TGF-β1 and VEGF were decreased,the mRNA expression level of TIMP-1 in nasal mucosa was increased,the mRNA expression level of MMP-9 was decreased,and the protein expression levels of Wnt-1 andβ-catenin in nasal mucosa were decreased(all P<0.05)in a dose-dependent manner.HE staining results showed that the morphology of nasal mucosa cells was significantly improved.Conclusion Yiqi Tongqiao Prescription inhibits the activity of MMP-9 through the endogenous regulation of TIMP-1 to maintain the homeostasis of extracellular matrix in nasopharyngeal carcinoma,and inhibits the abnormal activation of Wnt/β-catenin signaling pathway and reduces the release of TGF-β1 and VEGF to reduce tumor angiogenesis,thereby playing a role in tumor inhibition for nude mice with nasopharyngeal carcinoma.