中药药理与临床2025，Vol.41Issue(9)：33-39,7.
加味丹玄口康调节YAP/TAZ信号通路对槟榔碱诱导的口腔黏膜下纤维化的体内外实验研究
Jiawei Danxuan Koukang(加味丹玄口康)Regulates YAP/TAZ Signaling Pathway to Alleviate Arecoline-Induced Oral Submucosal Fibrosis in vivo and in vitro
摘要
Abstract
Objective:To investigate the effect and mechanism of Jiawei Danxuan Koukang(加味丹玄口康)in alleviating arecoline-induced oral submucosal fibrosis by regulating the Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ)signaling pathway in vivo and in vitro and delay the progression of oral submucosal fibrosis and carcinogenesis.Methods:(1)SD rats were modeled for OSF and then randomly assigned into model control,Jiawei Danxuan Koukang(6 g/kg and 12 g/kg),and YAP inhibitor verteporfin(10 mg/kg)groups,with 12 rats in each group.In addition,12 rats were selected as the normal control group.After 4 weeks of intervention,the changes of rat mouth opening were detected.Hematoxylin-eosin and Masson staining were used to observe the pathological changes of oral buccal mucosa,and the epithelial peg length and the collagen volume fraction were measured.Western blot and qRT-PCR were employed to determine the protein and mRNA levels,respectively,of E-cadherin,collagen Ⅰ,vimentin,and YAP/TAZ pathway-related factors in rat oral buccal mucosa.Enzyme-linked immunosorbent assay(ELISA)was employed to determine the serum levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,and transforming growth factor(TGF)-β1.(2)Thirty SD rats were randomly assigned into a blank control group and a Jiawei Danxuan Koukang(12 g/kg)group,and then the drug-containing serum was prepared.HaCaT cells were cultured in vitro,and the Cell Counting Kit-8(CCK-8)method was used to investigate the effects of different concentrations of arecoline,Jiawei Danxuan Koukang-containing serum,and verteporfin on the survival rate of HaCaT cells.The cells were classified into blank control,model control,10%Jiawei Danxuan Koukan(24 g/kg)-containing serum,and verteporfin groups.Western blot and qRT-PCR were employed to determine the protein and mRNA levels,respectively,of E-cadherin,collagen Ⅰ,vimentin,and YAP/TAZ pathway-related factors in each group of cells.ELISA was employed to determine the levels of TNF-α,IL-1β,and TGF-β1 in each group of cells.Results:(1)In animal experiments,compared with the normal control group,the model control group showed a reduced mouth opening degree,epithelial atrophy,inflammatory cell infiltration,in-creased collagen fiber deposition,severe pathological damage,elevated levels of TNF-α,IL-1β,and TGF-β1,up-regulated protein and mRNA levels of collagen Ⅰ,vimentin,YAP,and TAZ,and down-regulated protein and mRNA levels of E-cadherin(P<0.05 or P<0.01).Compared with model control group,Jiawei Danxuan Koukang at different doses and verteporfin significantly increased the mouth opening degree,led to thickened epithelium and reduced inflammatory cells and collagen fibers,and alleviated the pathological damage.In addition,Jiawei Danxuan Koukang at different doses and verteporfin lowered the levels of TNF-α,IL-1β and TGF-β1,down-regulated the protein and mRNA levels of collagen Ⅰ,vimentin,YAP,and TAZ,and up-regulated the protein and mRNA levels of E-cadherin(P<0.05 or P<0.01).(2)In cell ex-periments,according to the results of CCK-8,75 μg/mL ANE,10%Jiawei Danxuan Koukang(24 g/kg)-containing serum,and 200 nmol/L verteporfin were selected for cell treatment.Compared with the blank control group,the model control group showed elevated levels of TNF-α,IL-1β,and TGF-β1,significantly up-regulated protein and mRNA levels of collagen Ⅰ,vimentin,YAP,and TAZ,and significantly down-regu-lated protein and mRNA levels of E-cadherin.Compared with the model control group,10%Jiawei Danxuan Koukang(12 g/kg)-containing serum and verteporfin lowered the levels of TNF-α,IL-1β,and TGF-β1,down-regulated the protein and mRNA levels of collagen Ⅰ,vimen-tin,YAP,and TAZ,and up-regulated the protein and mRNA levels of E-cadherin(P<0.05 or P<0.01).Conclusion:Jiawei Danxuan Kou-kang can inhibit the inflammatory response of oral mucosal cells in vitro and in vivo by down-regulating the expression of YAP/TAZ signaling pathway,thereby alleviating oral submucosal fibrosis.关键词
加味丹玄口康/口腔黏膜下纤维化/Yes相关蛋白/具有PDZ结合基序的转录辅激活因子/槟榔碱Key words
Jiawei Danxuan Koukang(加味丹玄口康)/Oral submucous fibrosis/Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ)/Arecoline引用本文复制引用
李群,刘寻,朱紫冰,周婷婷,肖艳波,谭劲..加味丹玄口康调节YAP/TAZ信号通路对槟榔碱诱导的口腔黏膜下纤维化的体内外实验研究[J].中药药理与临床,2025,41(9):33-39,7.基金项目
国家自然科学基金面上项目(编号:82374530) （编号:82374530）
湖南省教育厅科研项目自然科学类(编号:24B0340) （编号:24B0340）
湖南中医药大学校院联合基金(编号:2024XYLH016) （编号:2024XYLH016）
湖南中医药大学口腔医学一级学科开放基金重点项目(编号:2019KQYX09). （编号:2019KQYX09）
