牙龈卟啉单胞菌脂多糖促进牙龈成纤维细胞的自噬  

Lipopolysaccharide of Porphyromonas gingivalis promotes the autophagy of human gingival fibroblasts

作　　者：刘瑜[1] 李树锦[2] 张森林[3] 操小马[1] 张艳梅[1] 

机构地区：[1]合肥市第二人民医院口腔科,安徽合肥230011 [2]合肥市第二人民医院检验科,安徽合肥230011 [3]南京军区总医院口腔科,江苏南京210002

出　　处：《细胞与分子免疫学杂志》2017年第3期315-319,共5页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的研究牙龈卟啉单胞菌脂多糖(Pg-LPS)对人牙龈成纤维细胞(HGF)自噬功能的影响。方法以10μg/mL Pg-LPS刺激HGF 12 h或24 h,采用雷帕霉素作为阳性对照,Western blot法检测微管相关蛋白1轻链3B(LC3B)的表达,间接免疫荧光技术检测自噬体的分布;同时使用MitoSOX Red荧光探针标记线粒体活性氧(mtROS)、间接免疫荧光法检测自噬体,观察Pg-LPS处理后HGF mtROS与线粒体自噬水平;分别以叔丁基-4-羟基茴香醚(BHA)、N-乙酰半胱氨酸(NAC)以及辅酶Q10(CoQ10)阻断ROS,Western blot法检测Pg-LPS刺激后LC3B的表达量。结果 Pg-LPS处理后LC3BⅡ/LC3BⅠ的比值与自噬体数量均显著升高,并且24 h处理组高于12 h处理组,同时mtROS生成量与线粒体自噬均明显增加。此外CoQ10可有效降低Pg-LPS诱导的HGF自噬。结论 Pg-LPS在HGF中通过触发mtROS活化自噬,并且自噬参与受损线粒体的降解以维持细胞内环境稳态。Objective To investigate the impact of lipopolysaccharide of Porphyromonas gingivalis（Pg-LPS） on the autophagy of human gingival fibroblasts（HGFs）. Methods Firstly,HGFs was stimulated with 10 μg/mL Pg-LPS for 12 hours or 24 hours. Rapamycin was used as a positive control. The expression of LC3 B was detected by Western blotting and the distribution of autophagosomes was observed by indirect immunofluorescence staining. At the same time,mitochondrial ROS（mtROS） was labeled by MitoSOX Red. The levels of mtROS and mitochondrial autophagy were measured in HGFs after treated with Pg-LPS. Then,T-butyl-4（BHA）,N-acetylcysteine（NAC） and coenzyme Q10（CoQ10） were used separately to block the ROS and the expression of LC3 B in Pg-LPS-treated HGFs was tested by Western blotting. Results After treatment with Pg-LPS,the ratio of LC3BⅡ/LC3BⅠ and the number of autophagic cells significantly increased,and the increase in the 24-hour treatment group was the more obvious than that in the 12-hour treatment group. The mtROS production and mitochondrial autophagy were significantly promoted after Pg-LPS treatment. In addition,CoQ10 effectively reduced Pg-LPS-induced autophagy of HGFs. Conclusion Pg-LPS can induce the autophagy of HGFs by raising mtROS production,and autophagy is involved in the degradation of damaged mitochondria to maintain cellular homeostasis.

关 键 词：牙龈卟啉单胞菌 脂多糖 自噬 活性氧 

分 类 号：R392.33[医药卫生—免疫学] R781.52[医药卫生—基础医学]