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益气通窍方对鼻咽癌裸鼠的抑瘤作用及机制

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[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.
Anti-Tumor Effect and Mechanism of Yiqi Tongqiao Prescription on Nasopharyngeal Carcinoma in Nude Mice
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 factor β1(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 and β-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 and β-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.

Yiqi Tongqiao Prescriptionnasopharyngeal carcinomatissue inhibitor of metalloproteinase 1(TIMP-1)matrix metalloproteinase 9(MMP-9)transforming growth factor β1(TGF-β1)vascular endothelial growth factor(VEGF)Wnt-1/β-catenin pathwaynude mice

常雪岩、周凌、杨佳鹏、刘静丹

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黑龙江中医药大学,黑龙江哈尔滨 150040

黑龙江中医药大学附属第一医院,黑龙江哈尔滨 150040

益气通窍方 鼻咽癌 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) 转化生长因子β1(TGF-β1) 血管内皮细胞生长因子(VEGF) Wnt-1/β-连环蛋白(β-catenin)通路 裸鼠

国家自然科学基金委员会资助项目

81874495

2024

广州中医药大学学报
广州中医药大学

广州中医药大学学报

CSTPCD
影响因子:1.055
ISSN:1007-3213
年,卷(期):2024.41(9)