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在高糖条件下,脂多糖诱导的牙周膜细胞增殖抑制和凋亡会增强。

Lipopolysaccharide-induced suppression of periodontal ligament cell proliferation and apoptosis are strengthened under high glucose conditions.

作者信息

Liu Jianxin, Chen Shulan, Ren Weiwei, Liu Jianing, Yang Pishan, Chen Zhenggang, Zhang Qiang, Yang Fang

机构信息

Department of Reproductive Medicine, Affiliated Hospital of Qingdao University, Qingdao 266000, China.

Department of Stomatology, Qingdao Municipal Hospital, Qingdao 266011, China.

出版信息

Arch Oral Biol. 2017 Jul;79:70-76. doi: 10.1016/j.archoralbio.2017.01.007. Epub 2017 Jan 17.

Abstract

The present study aimed to investigate the effect of lipopolysaccharide (LPS) on the proliferation and apoptosis of human periodontal ligament (hPDL) cells under normal glucose or high glucose conditions. Primary cultures of hPDL cells were prepared from extracted premolars of patients. The cells were incubated with 0, 1, or 10μg/mL LPS under normal glucose (5.5mmol/L) or high glucose (25mmol/L) conditions for 24h or 48h. Cell proliferation was detected using a CCK-8 assay, and cell apoptosis was measured by Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining. BCL2 and BAX mRNA and protein levels were measured by real-time polymerase chain reaction and western blotting, respectively. LPS (10μg/mL) induced significant inhibition of cell proliferation and cell apoptosis, and a significant decrease in the BCL2/BAX ratio in the cells cultured with 5.5mmol/L glucose. These effects of LPS were increased significantly in cells treated with 25mmol/L glucose. Analysis of variance of the factorial design revealed that high glucose and LPS had a significant interaction for cell apoptosis, but not for cell proliferation. High glucose augmented LPS-induced hPDL cell apoptosis and cell proliferation inhibition. LPS and high glucose might interact to induce cell apoptosis.

摘要

本研究旨在探讨脂多糖(LPS)在正常葡萄糖或高葡萄糖条件下对人牙周膜(hPDL)细胞增殖和凋亡的影响。从患者拔除的前磨牙制备hPDL细胞原代培养物。将细胞在正常葡萄糖(5.5mmol/L)或高葡萄糖(25mmol/L)条件下与0、1或10μg/mL LPS孵育24小时或48小时。使用CCK-8法检测细胞增殖,通过Hoechst 33258染色和膜联蛋白V-异硫氰酸荧光素(FITC)/碘化丙啶(PI)双染色测量细胞凋亡。分别通过实时聚合酶链反应和蛋白质印迹法测量BCL2和BAX的mRNA和蛋白质水平。LPS(10μg/mL)在5.5mmol/L葡萄糖培养的细胞中诱导细胞增殖和细胞凋亡的显著抑制,以及BCL2/BAX比值的显著降低。在25mmol/L葡萄糖处理的细胞中,LPS的这些作用显著增强。析因设计的方差分析显示,高葡萄糖和LPS对细胞凋亡有显著交互作用,但对细胞增殖没有。高葡萄糖增强了LPS诱导的hPDL细胞凋亡和细胞增殖抑制。LPS和高葡萄糖可能相互作用诱导细胞凋亡。

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