牙龈卟啉单胞菌脂多糖促进牙龈成纤维细胞的自噬

发布时间：2018-04-03 00:33

  本文选题：牙龈卟啉单胞菌　切入点：脂多糖　出处：《细胞与分子免疫学杂志》2017年03期

【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。
[Abstract]:Objective to study the effect of Pg-LPSs from porphyromonas gingivalis on the autophagy of human gingival fibroblasts (HGF).Methods 10 渭 g/mL Pg-LPS was used to stimulate HGF for 12 h or 24 h. The expression of microtubule-associated protein 1 light chain 3BC3LC _ 3B was detected by Western blot, and the distribution of autophagy was detected by indirect immunofluorescence technique.At the same time, MitoSOX Red fluorescence probe was used to label mitochondrial reactive oxygen species (Ros) and mtROSs, and indirect immunofluorescence assay was used to detect autophagy. The levels of HGF mtROS and mitochondrial autophagy after Pg-LPS treatment were observed.The expression of LC3B after Pg-LPS stimulation was detected by blot method with blot and coenzyme Q10 CoQ10 with tert-#china_person0#-4-hydroxyanisole (BHA) and coenzyme Q10-CoQ10 (n-acetylcysteine).Results after Pg-LPS treatment, the ratio of LC3B 鈪，

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