绿原酸对牙龈卟啉单胞菌诱导牙龈上皮细胞自噬的调控作用及机制  

Regulation and mechanism of chlorogenic acid on autophagy of gingival epithelial cells induced by porphyromonas gingivalis

作　　者：吴珊 赵国廷 董振耀 马秀花 姚毅章 WU Shan;ZHAO Guoting;DONG Zhenyao;MA Xiuhua;YAO Yizhang(Department of Stomatology,The Fifth People′s Hospital of Qinghai Province,Xining 810007,China)

机构地区：[1]青海省第五人民医院口腔科,青海西宁810007

出　　处：《西部医学》2023年第12期1751-1756,共6页Medical Journal of West China

基　　金：青海省科技计划项目(2017-ZJ-707)。

摘　　要：目的探讨绿原酸(CGA)对牙龈卟啉单胞菌(P.g)诱导人牙龈上皮细胞(HGE)自噬的影响及作用机制。方法将HGE细胞分为对照组(Control组,做任何处理)、P.g组(使用P.g诱导HGE细胞建立细胞模型)、CGA组(同P.g组处理细胞,再分别用10.0、20.0、40.0 mg/L CGA)、CGA+AMPK激活剂(AICAR)组(同P.g组处理细胞,再用40.0 mg/L CGA+1.0 nmol/L AICAR)。MTT法检测细胞增殖。分别检测HGE细胞凋亡及LC3Ⅱ、LC3I、Beclin-1、P62、p-ULK1、ULK1、p-AMPK、AMPK蛋白表达。结果10~60 mg/L的CGA均可显著抑制P.g诱导的HGE细胞增殖,选择浓度为10.0、20.0、40.0 mg/L的CGA进行后续实验。与Control组比较,P.g组LC3Ⅱ/I、Beclin-1、p-ULK1、p-AMPK蛋白表达水平显著升高,P62表达水平显著降低(P<0.05);与P.g组比较,浓度为10.0、20.0、40.0 mg/L的CGA处理的HGE细胞凋亡率显著升高(P<0.05),LC3Ⅱ/I、Beclin-1、p-ULK1、p-AMPK蛋白表达水平显著降低,P62表达水平显著升高(P<0.05),且呈药物浓度依赖性;AICAR可部分逆转CGA对P.g诱导的HGE细胞自噬的抑制作用(P<0.05)。结论CGA通过抑制AMPK/ULK1通路,抑制P.g诱导的人牙龈上皮细胞自噬,促进细胞凋亡。Objective To investigate the influence and mechanism of chlorogenic acid(CGA)on autophagy induced by porphyromonas gingivalis(Pg)in human gingival epithelial cells(HGE).Methods HGE cells were cultured in vitro,and Pg was used to induce HGE cells to establish a cell model.Pg-induced HGE cells were treated with 5-60 mg/L CGA,respectively.Cell proliferation was detected by MTT assay,HGE cells were grouped into Control group,Pg group,CGA group(drug concentrations of 10.0,20.0,40.0 mg/L,respectively),and CGA+AMPK activator(AICAR)group(40.0 mg/L CGA+1.0 nmol/L AICAR),the apoptosis of HGE cells and the protein expressions of LC3Ⅱ,LC3I,Beclin-1,P62,p-ULK1,ULK1,p-AMPK and AMPK were detected respectively.Results CGA at 10-60 mg/L could greatly inhibit the proliferation of HGE cells induced by Pg.CGA at concentrations of 10.0,20.0,and 40.0 mg/L were selected for subsequent experiments.Compared with the control group,the protein expression levels of LC3Ⅱ/I,Beclin-1,p-ULK1 and p-AMPK in the Pg group were greatly increased,and the expression level of P62 was greatly decreased(P<0.05);compared with the Pg group,the apoptosis rate of HGE cells treated with CGA at concentrations of 10.0,20.0,and 40.0 mg/L was greatly increased(P<0.05),the protein expression levels of LC3Ⅱ/I,Beclin-1,p-ULK1,and p-AMPK were greatly decreased,and the expression level of P62 was greatly increased(P<0.05),which were drug concentration-dependent;AICAR could partially reverse the inhibitory effect of CGA on Pg-induced autophagy in HGE cells(P<0.05).Conclusion CGA inhibits Pg-induced autophagy of human gingival epithelial cells and promotes cell apoptosis by inhibiting the AMPK/ULK1 pathway.

关 键 词：绿原酸 AMPK/ULK1信号通路 牙龈卟啉单胞菌 牙龈上皮细胞 自噬 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R780.2[医药卫生—基础医学]