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Ku70 在炎性牙髓干细胞凋亡中的作用。

Role of Ku70 in the apoptosis of inflamed dental pulp stem cells.

机构信息

The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, Hubei, People's Republic of China.

出版信息

Inflamm Res. 2018 Sep;67(9):777-788. doi: 10.1007/s00011-018-1167-2. Epub 2018 Jul 14.

Abstract

AIM

The study aimed to investigate the effects of DNA repair proteins on cell apoptosis in human DPSCs during inflammation.

METHODS

Lipopolysaccharide (LPS) was used to stimulate inflammation in dental pulp in vivo and in vitro. We identified the activation of DSB response and DNA repair proteins in inflamed pulp tissue and in LPS-treated human DPSCs. Then we transfected the cells with Ku70 (a key protein involved in NHEJ) siRNA and detected the expression changes of γ-H2A.X, DNA repair proteins and cell apoptosis.

RESULTS

Immunohistochemical staining showed that at 4 and 6 days of pulpitis the expression of Ku70 and γ-H2A.X significantly increased. The levels of γ-H2A.X, Ku70, Xrcc4, and Rad51 increased considerably in the LPS-treated DPSCs. Furthermore, decreased expression of Ku70 could increase the number of γ-H2A.X foci, apoptotic cells and reduce cell viability in DPSCs.

CONCLUSIONS

The results indicate that NHEJ pathway was the main mechanism involved in DNA damage response induced by repeated LPS stimulation in DPSCs. Meanwhile, the findings suggested that Ku70 serves importantly in the apoptosis of DPSCs in the inflammatory environment.

摘要

目的

本研究旨在探讨 DNA 修复蛋白在炎症状态下人牙髓间充质干细胞(DPSCs)细胞凋亡中的作用。

方法

体内和体外实验均使用脂多糖(LPS)刺激牙髓炎症。我们鉴定了炎症牙髓组织和 LPS 处理的人 DPSCs 中 DSB 反应和 DNA 修复蛋白的激活情况。然后,我们用 Ku70(一种参与非同源末端连接(NHEJ)的关键蛋白)siRNA 转染细胞,并检测 γ-H2A.X、DNA 修复蛋白和细胞凋亡的表达变化。

结果

免疫组织化学染色显示,在牙髓炎的第 4 天和第 6 天,Ku70 和 γ-H2A.X 的表达显著增加。LPS 处理后的 DPSCs 中 γ-H2A.X、Ku70、Xrcc4 和 Rad51 的水平显著升高。此外,Ku70 的表达下调可增加 γ-H2A.X 焦点、凋亡细胞的数量,并降低 DPSCs 的细胞活力。

结论

结果表明,NHEJ 途径是 LPS 反复刺激 DPSCs 引起的 DNA 损伤反应的主要机制。同时,研究结果提示 Ku70 在炎症环境中 DPSCs 的凋亡中起着重要作用。

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