Liaoning Provincial Key Laboratory of Oral Diseases, Department of Periodontology, School and Hospital of Stomatology, China Medical University, Shenyang, China.
Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China.
Oral Dis. 2021 Oct;27(7):1775-1788. doi: 10.1111/odi.13696. Epub 2020 Nov 5.
This paper aims to study the effect of the active form of vitamin D (calcitriol) on the internalized Porphyromonas gingivalis in macrophages and to assess the role of autophagy during this process.
Quantitative RT-PCR and bacteria culture were used to quantify live P. gingivalis internalized into U937-derived macrophages. Western blot assays were performed to detect the effect of P. gingivalis and calcitriol on autophagy in macrophages. Transmission electron microscope was used to observe the effect of calcitriol on the status of internalized P. gingivalis. Colocalization of P. gingivalis with the autophagosome and lysosome markers was observed by confocal laser scanning microscopy.
Calcitriol caused a dose-dependent decrease in live P. gingivalis numbers and promoted both the endogenous and P. gingivalis-induced autophagy in macrophages. Calcitriol significantly promoted the destruction of P. gingivalis and the colocalization of P. gingivalis with autophagosome and lysosome markers. Conversely, with 3-MA, live P. gingivalis numbers in macrophages increased significantly and inhibition effect of calcitriol on the number of live P. gingivalis was attenuated.
In U937-derived macrophages, calcitriol may promote colocalization of P. gingivalis with autophagosomes and lysosomes, namely autophagy process, to degrade live P. gingivalis.
本研究旨在探讨活性维生素 D(骨化三醇)对巨噬细胞内化的牙龈卟啉单胞菌的影响,并评估自噬在此过程中的作用。
采用定量 RT-PCR 和细菌培养技术来量化 U937 衍生的巨噬细胞内化的活牙龈卟啉单胞菌数量。通过 Western blot 实验检测牙龈卟啉单胞菌和骨化三醇对巨噬细胞自噬的影响。采用透射电子显微镜观察骨化三醇对内化的牙龈卟啉单胞菌状态的影响。通过共聚焦激光扫描显微镜观察牙龈卟啉单胞菌与自噬体和溶酶体标志物的共定位情况。
骨化三醇呈剂量依赖性降低活牙龈卟啉单胞菌数量,并促进巨噬细胞内源性和牙龈卟啉单胞菌诱导的自噬。骨化三醇显著促进了牙龈卟啉单胞菌的破坏和与自噬体和溶酶体标志物的共定位。相反,用 3-MA 处理后,巨噬细胞内活牙龈卟啉单胞菌数量显著增加,骨化三醇对活牙龈卟啉单胞菌数量的抑制作用减弱。
在 U937 衍生的巨噬细胞中,骨化三醇可能通过促进牙龈卟啉单胞菌与自噬体和溶酶体的共定位,即自噬过程,降解活牙龈卟啉单胞菌。