Low-intensity pulsed ultrasound activates autophagy in periodontal ligament cells in the presence or absence of lipopolysaccharide
Low-intensity pulsed ultrasound activates autophagy in periodontal ligament cells in the presence or absence of lipopolysaccharide
复制标题
在存在或不存在脂多糖的情况下，低强度脉冲超声激活牙周膜细胞的自噬
DOI：
10.1016/j.archoralbio.2020.104769
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发表时间：
2020-09-01
影响因子：
3
通讯作者：
Song, Jinlin
中科院分区：
文献类型：
--
作者：
Li, Yao;Sun, Chengjun;Song, Jinlin
目的：本研究旨在确定低强度脉冲超声（LIPUS）是否激活人牙周膜细胞（PDLCs）中的自噬，而与脂多糖无关。设计：六组被设计：对照、LIPUS、脂多糖、LIPUS +脂多糖、LIPUS+3-甲基腺嘌呤、LIPUS +脂多糖+3-甲基腺嘌呤。LIPUS预处理PDLCs 2 h，脂多糖处理不同时间。采用Real-time PCR和Western-blot检测自噬基因Beclin-1和LC 3的mRNA和蛋白表达水平。透射电镜观察自噬体的形态。结果：与未处理组相比，LIPUS预处理组4 h和8h时LC 3和Beclin-1的mRNA表达水平显著升高（P <0.05），LC 3 - 1的蛋白表达水平显著升高（P< 0.05）。Beclin-1在4 h、8h、16 h表达量显著高于对照组（P< 0.05）。LIPUS预处理和脂多糖处理8 h后，LC 3-. Beclin-1蛋白表达水平明显高于对照组（P< 0.05）。进一步用3-甲基腺嘌呤处理后，Beclin-1蛋白表达较LIPUS+脂多糖组降低（P< 0.05），而LC 3-。蛋白质表达则没有。LIPUS+3-Methyladenine组和LIPUS+lipopolysaccharide+3-Methyladenine组未见自噬体。LIPUS预处理和脂多糖处理36 h后，IL-6表达较脂多糖组明显降低（P< 0.05）。结论：LIPUS可促进PDLCs自噬，且与脂多糖无关。自噬可能参与了LIPUS对PDLCS的抗炎作用机制。
Objective: This study aims to determine if low-intensity pulsed ultrasound (LIPUS) activates autophagy in human periodontal ligament cells (PDLCs) irrespective of lipopolysaccharide.Design: Six groups were designed: control, LIPUS, lipopolysaccharide, LIPUS + lipopolysaccharide, LIPUS+3-Methyladenine, LIPUS + lipopolysaccharide+3-Methyl- adenine. LIPUS pretreated PDLCs for 2 h and lipopolysaccharide treated for different times. Real-time PCR and Western-blot were performed to evaluate mRNA and protein expression levels of autophagic genes Beclin-1 and LC3 respectively. A transmission electronic microscope was used to observe the autophagosome. ELISA was used to test interleukin-6 expression.Results: Compared with the non-treatment, LIPUS pretreatment increased mRNA expression levels of LC3 (P< 0.05) and Beclin-1 (P< 0.05) at 4 h and 8 h, and enhanced the protein expression levels of LC3-. at 8 h (P< 0.05) and Beclin-1 at 4 h, 8 h and 16 h(P< 0.05). After LIPUS pretreatment and lipopolysaccharide treatment for 8 h, LC3-. and Beclin-1 protein expression levels were elevated (P< 0.05) compared with the control. Following further treatment by 3-Methyladenine, Beclin-1 protein expression was decreased (P< 0.05) compared with the LIPUS plus lipopolysaccharide group, but LC3-. protein expression was not. Autophagosomes were not found in the LIPUS+3-Methyladenine and LIPUS+lipopolysaccharide+3-Methyladenine groups. After LIPUS pretreatment and lipopolysaccharide treatment for 36 h, intreleukin-6 expression was decreased (P< 0.05) compared with the lipopolysaccharide group. However, after addition of 3-Methyladenine, intreleukin-6 expression was elevated (P< 0.05) compared with the LIPUS +lipopolysaccharide group.Conclusions: LIPUS can promote autophagy in PDLCs irrespective of lipopolysaccharide. Autophagy might be involved in LIPUS anti-inflammatory mechanism in PDLCS.