摘要
目的探索扶正活血解毒方对槟榔碱提取物(areca nut extract,ANE)诱导的口腔黏膜下纤维化(oral submucous fibrosis,OSF)Wnt/β-catenin信号通路的作用机制。方法体外培养大鼠口腔黏膜上皮细胞(epithelial cell,EC),分为正常组、模型组、中药高/中/低剂量组和IWR-1组。以ANE为诱导剂,扶正活血解毒方含药血清为干预药物,以Wnt/β-catenin信号通路抑制剂IWR-1为阳性对照物,采用CCK8检测EC细胞增殖;PCR检测Wnt1、β-catenin、Axin、cylinD1基因的表达;Western blot检测Wnt1、β-catenin、Axin、cylinD1蛋白的表达。结果与正常组相比,模型组EC增殖水平提高,Wnt1、β-catenin、cylinD1基因及蛋白表达增加,Axin基因及蛋白表达降低(P<0.05);与模型组相比,中药高、中剂量组EC增殖水平降低,Wnt1、β-catenin、cylinD1基因及蛋白表达降低,Axin基因及蛋白表达升高(P<0.05);与模型组相比,IWR-1组EC增殖水平降低,Wnt1、β-catenin、cylinD1基因及蛋白表达降低,Axin基因及蛋白表达升高(P<0.05)。结论扶正活血解毒方可通过调控Wnt/β-catenin信号通路,抑制ANE刺激的EC增殖,逆转OSF癌前病变的形成。
Objective To explore the mechanism of Fuzheng Huoxue Jiedu Prescription on Wnt/β-catenin signaling pathway induced by areca nut extract(ANE)in oral submucosal fibrosis.Methods The rat oral mucosa epithelial cells(ECs)cultured in vitro were assigned into a normal group,a model group,a Chinese materia medica high,medium and low dose group,a IWR-1 group.ANE was inducer,and Fuzheng Huoxue Jiedu Prescription-containing serum was intervention medicine.Wnt/β-catenin signaling pathway inhibitor IWR-1 was the positive control medicine.The CCK8 was used to test EC cell proliferation;PCR was used to detect Wnt1,β-catenin,Axin,cylinD1 gene expression;Western Blot was used to detect Wnt1,β-catenin,Axin,cylinD1 protein expression.Results Compared with the normal group,ECs proliferation levels of the model group increased,and Wnt1,β-catenin,cylinD1 gene and protein expression increased.Axin gene and protein expression decreased(P<0.05);Compared with model group,ECs proliferation levels of Chinese materia medica high and medium dose groups were reduced,and Wnt1,β-catenin,cylinD1 gene and protein expression were reduced.Axin gene and protein expression increased(P<0.05);Compared with the model group,ECs proliferation level of the IWR-1 group was reduced,and Wnt1,β-catenin,cylinD1 gene and protein expression were reduced.Axin gene and protein expression increased(P<0.05).Conclusion Fuzheng Huoxue Jiedu Prescription can inhibit the proliferation of EC cells stimulated by ANE,and reverse the formation of OSF precancerous lesions through regulating Wnt/β-catenin signaling pathway.
作者
谢赛飞
谭劲
郑玲
朱可可
周领航
陈明
XIE Saifei;TAN Jin;ZHEN Ling;ZHU Keke;ZHOU Linghang;CHEN Ming(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China)
出处
《湖南中医药大学学报》
CAS
2020年第11期1305-1309,共5页
Journal of Hunan University of Chinese Medicine
基金
国家自然科学基金项目(81874496)
湖南省中医药管理局课题(201808)
湖南中医药大学研究生创新课题(2019CX73)。
