Effects Of Jiawei Danxuankoukang On KLF4 Expression Induced By ANE In Oral Mucosal Epithelial Cells

Objective: This study intends to intervene in oral mucosa epithelial cells of rats by 50?g/ml arecoline and low and high dose Jiawei Danxuankoukang,to simulate the effect of arecoline on epithelial cells in saliva when chewing betel nut and drug intervention to detect rats The expression of KLF4 in oral mucosal epithelial cells,and by observing and recording the biological behavior of epithelial cell lines and changes in related genes ?-catenin,to clarify the function of KLF4 in the development and canceration of OSF.Method:1.Preparation of Jiawei Danxuankoukang drug serum:Fifteen adult healthy SD rats were randomly divided into three groups: A: normal group,B:Jiawei Danxuankoukang low,C: high dose group.Five rats in each group were given intragastric administration with Distilled water and Jiawei Danxuankoukang 4ml / kg and 12 ml / kg twice a day for 7 days.Blood was collected from the carotid artery under sterile conditions and prepared into a culture medium containing 10% rat serum for later use.2.Cell culture:Culture primary rat oral mucosal epithelial cells.Incubate in a 5% CO2,37 ? incubator,take the logarithmic growth phase cells for experiment.3.Cell passaging and intervention grouping:Group A(NC Group): 10%Normal rat drug serum + 2% complete epithelial cell+ Distilled water,Group B(ANE Group): 50 ug / ml ANE induced rat oral mucosal epithelial cells + 10% Normal rat drug serum + 2% complete epithelial cell culture fluid,Group C(LDM Group): 50 ug / ml ANE induced rat oral mucosal epithelial cells + low-dose Jiawei Danxuankoukang drug serum + 2%complete epithelial cell culture medium,Group D(HDM Group): 50 ug / ml ANE induced rat oral mucosal epithelial cells + high-dose Jiawei Danxuankoukang drug serum + 2% complete culture medium of epithelial cells.Incubate for 24 h.4.Index detection:(1)Observe the changes of cell morphology and cell density under the microscope;(2)Q-PCR technology to detect the transcription of KLF4 and ?-catenin;(3)Western blot to detect the quantitative expression of KLF4 and ?-catenin protein.Result:1.Microscopic observation: Compared with the normal serum group,the B,C,and D groups after the intervention of arecoline have reduced cell viability,decreased density,and changed cell morphology.In group D,the cell viability and cell density were improved compared with group B without Jiawei Danxuankoukang,and the change was correlated with the dose.2.Western blot results: Compared with the normal serum group,the expression of KLF4 decreased and the expression of ?-catenin increased in groups B,C,and D after the intervention of arecoline.The differences are statistically significant.Compared with the group B and the group B without the Jiawei Danxuankoukang,the KLF4 expression was increased,and the ?-catenin expression was decreased,and the differences were statistically significant,and were related to the dose.Sex.3.Results of Q-PCR: Compared with the normal serum group,the B,C,and D groups after the intervention of arecoline reduced KLF4 expression and increased ?-catenin expression.The differences are statistically significant.Compared with the B group without the Jiawei Danxuankoukang group C and D,the expression of KLF4 was increased,and the ?-catenin expression was decreased,and the high-dose group was different from the arecoline-induced group.Statistically significant.Conclusion:1.50 ?g / ml Arecoline acts on rat oral mucosal epithelial cells.Compared with the normal serum group,the Arecoline-induced group reduces the expression of KLF4;the cell viability per unit area decreases,the number of cells decreases,and the cell morphology changes,The intercellular space increases,and abnormal mitotic figures appear.2.Jiawei Danxuankoukang can antagonize the cytotoxicity of arecoline on mucosal epithelial cells.Compared with the arecoline-induced group,KLF4 expression is enhanced,cell viability,proliferative capacity,and cell morphology are improved.3.KLF4 may be related to oral submucosal fibrosis and carcinogenesis.KLF4 inhibits the expression of Wnt / ?-catenin signaling pathway may be one of the mechanisms by which Jiawei Danxuankang inhibits the occurrence and development of OSF carcinogenesis.