目的:关于氧化应激诱导的衰老参与根尖周炎的研究知之甚少。这里,我们探讨了其在根尖周病变中的分子机制。
方法:随机选择10例根尖周围肉芽肿和5例。进行免疫组织化学分析以检测衰老相关因子聚合酶I和转录释放因子(PTRF)与Akt/FoxO1信号传导之间的表达和相关性。用LY294002预处理的人牙周膜细胞(hPDLCs)暴露于H2O2诱导的氧化应激条件下,然后细胞增殖,衰老,凋亡,和相关的信号通过EdU标记进行评估,β-半乳糖苷酶测定,RT-qPCR,和蛋白质印迹分析,分别。
结果:聚合酶I和转录物释放因子和Akt/FoxO1信号在根尖肉芽肿中的表达频率高于根尖肉芽肿。值得注意的是,根性囊肿中的细胞显示Akt激活,FoxO1磷酸化,和细胞质易位。体外,在hPDLCs中观察到显著的H2O2诱导的衰老。LY294002,一种PI3K抑制剂,减弱衰老的表达水平(Klotho,P16INK4),凋亡(坏,Fas),磷酸化Akt,和磷酸化的FoxO1;然而,不影响细胞增殖。
结论:我们的数据表明衰老存在于临床根尖周病变中,Akt/FoxO1信号参与H2O2诱导的细胞衰老,可以作为潜在的治疗靶点。
There is little knowledge about oxidative stress-induced senescence involvement in apical periodontitis. Here, we explored its molecular mechanism in periapical lesions.
Ten cases of radicular cysts and five cases of periapical granulomas were randomly selected. Immunohistochemical analysis was performed to detect the expression and correlation between Senescence-associated factor polymerase I and transcript release factor (PTRF) and Akt/FoxO1 signaling. Human periodontal ligament cells (hPDLCs) pretreated with LY294002 were exposed to H2O2-induced oxidative stress conditions and then cell proliferation, senescence, apoptosis, and associated signaling were evaluated by EdU labeling, β-galactosidase assay, RT-qPCR, and western blot analysis, respectively.
Polymerase I and transcript release factor and Akt/FoxO1 signaling were more frequently expressed in the radicular cyst than in periapical granulomas. Notably, cells in radicular cysts showed Akt activation, FoxO1 phosphorylation, and cytoplasmic translocation. In vitro, prominent H2O2-induced senescence was observed in hPDLCs. LY294002, a PI3K inhibitor, attenuated the expression levels of senescence (Klotho, P16INK4), apoptosis (Bad, Fas), phosphorylated Akt, and phosphorylated FoxO1; however, did not affect cell proliferation.
Our data indicated that senescence is present in clinical periapical lesions, and Akt/FoxO1 signaling is involved in the H2O2-induced cellular senescence, which could serve as a potential therapeutic target.