基于痤疮丙酸杆菌诱导的细胞炎症模型研究温泉水的抗炎作用  

Study on the anti-inflammatory effect of thermal spring water based on a model of Propionibacterium acnes-induced cellular inflammation

作　　者：邹粤 张莹莹 杨芬 李红霞 张保超 徐义航 王瑞雪 裴晓方 ZOU Yue;ZHANG Ying-ying;YANG Fen;LI Hong-xia;ZHANG Bao-chao;XU Yi-hang;WANG Rui-xue;PEI Xiao-fang(West China School of Public Health/West China Fourth Hospital,Sichuan University,Chengdu,Sichuan 610041,China;不详)

机构地区：[1]四川大学华西公共卫生学院(华西第四医院),四川成都610041 [2]四川大学华西口腔医学院 [3]重庆市大渡口区疾病预防控制中心

出　　处：《现代预防医学》2024年第22期4191-4198,共8页Modern Preventive Medicine

基　　金：宜宾市科技计划资助项目(2021ZYSF006)。

摘　　要：目的构建痤疮丙酸杆菌(Propionibacterium acnes,P.acnes)诱导的细胞炎症模型研究某温泉水的抗炎作用。方法使用人永生化角质形成细胞(human immortalized keratinocytes,HaCaT)进行试验。将温泉水与细胞共孵育,用CCK-8法检测细胞存活率,筛选温泉水干预时间;将不同浓度P.acnes与细胞作用不同时间,检测细胞存活率和计算P.acnes的50%抑制浓度(IC_(50)),并检测IL-6含量,确定造模条件;温泉水浸泡细胞0.5 h后,再加入P.acnes共培养作为预防干预组,或先将P.acnes与细胞共培养,再加入温泉水浸泡细胞0.5 h作为治疗干预组,以CCK-8法、ELISA和RT-PCR分别检测细胞存活率、促炎细胞因子及其基因以及炎症通路相关基因表达的水平。同时设置生理盐水、自来水、纯水组以及模型和空白对照。结果温泉水和生理盐水作用0.5 h时,细胞存活率分别为92.7%和99.09%,将0.5 h作为后续实验的干预时间;OD_(600)值0.25的P.acnes与细胞共孵育5 h作为建模条件。预防式干预,仅温泉水组细胞的IL-1β、IL-8分泌量、IL-6、TNF-α、IL-8及TLR2、NF-κB基因的相对表达量均低于模型组(P<0.05),且明显低于生理盐水组。治疗式干预,温泉水组细胞的IL-6、IL-1β、TNF-α、IL-8含量以及IL-6、IL-1β、TNF-α、TLR2和NF-κB基因表达水平均低于模型组,其中IL-6、IL-1β、TNF-α含量以及IL-6、IL-1β、IL-8、TLR2、NF-κB基因表达水平也比生理盐水组的明显降低(P<0.05)。结论该温泉水可能通过下调TLR2和NF-κB炎症信号通路基因,降低促炎细胞因子分泌实现其抗炎作用。Objective To construct a Propionibacterium acnes(P.acnes)-induced cellular inflammation model and to study the anti-inflammatory effect of a thermal water.Methods Utilize human immortalized keratinocytes(HaCaT)was used in the experimentation.Co-incubate the cells with thermal spring water and employ the CCK-8 assay to assess cell viability,screening for the optimal intervention time for the thermal spring water.Expose the cells to varying concentrations of P.acnes for different times,measure cell viability,calculate the 50%inhibitory concentration(IC_(50))of P.acnes,and determine the model induction conditions by testing IL-6 levels.The cells were soaked in thermal spring water for 0.5 hours,followed by co-culturing with P.acnes as the preventive intervention group,or co-culturing with P.acnes first and then soaking the cells in hot spring water for 0.5 hours as the therapeutic intervention group.The levels of cell viability,pro-inflammatory cytokines,and gene expression of inflammation pathways were assessed using the CCK-8 assay,ELISA,and RT-PCR,respectively.Concurrently,control groups with saline solution,tap water,and pure water,along with model and blank controlswere set up.Results After treating the cells with thermal spring water and physiological saline for 0.5 hours,cell viabilities were 92.7%and 99.09%,respectively,which led to the selection of 0.5 hours as the intervention time for subsequent experiments.P.acnes at an OD_(600) of 0.25 was co-incubated with the cells for 5 hours to establish the model.In the preventive intervention,the secretion levels of IL-1β and IL-8,as well as the relative expression levels of IL-6,TNF-α,IL-8,and the genes TLR2 and NF-κB in the cells of the thermal spring water group,were all lower than those in the model group(P<0.05),and significantly lower than those in the saline solution group.In the therapeutic intervention,the contents of IL-6,IL-1β,TNF-α,and IL-8,as well as the expression levels of the genes IL-6,IL-1β,TNF-α,TLR2,and NF-κB in the thermal spring water

关 键 词：温泉水 痤疮丙酸杆菌 HACAT 抗炎作用 

分 类 号：R-33[医药卫生]