昼夜节律时钟蛋白 Bmal1 对实验性牙周炎相关肾损伤的影响

Haonan Ma, Qiong Li, Yaqi Shang, Xirui Xin, Xinchan Liu, Zhou Wu, Weixian Yu
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摘要

方法将12只雄性Wistar大鼠随机分为对照组和牙周炎组(每组6只)。用正畸结扎线结扎牙周炎大鼠上颌两侧的第一颗上颌磨牙,而对照组不采取任何干预措施。8 周后,评估两组大鼠的临床牙周参数,包括探诊深度、出血指数和牙齿活动度。对大鼠的上颌骨进行显微 CT 扫描和三维图像重建,以评估牙槽骨吸收情况。使用苏木精-伊红(HE)和周期性酸-希夫(PAS)染色法对牙周组织和肾组织进行组织病理学观察。肾功能指标(如肌酐、白蛋白和血尿素氮水平)和氧化应激指标(包括超氧化物歧化酶、谷胱甘肽和丙二醛水平)均采用生化检测试剂盒进行测量。MitoSOX 红染色用于检测肾脏中活性氧(ROS)的含量。使用实时定量聚合酶链反应(RT-qPCR)和免疫组化染色法评估了大鼠肾组织中 Bmal1、核因子红细胞 2 相关因子 2(Nrf2)和血红素加氧酶 1(HO-1)的基因和蛋白表达水平。通过 HE 和 PAS 染色进行的组织学检查显示,牙周炎组大鼠的肾组织受到了严重的组织病理学损伤。肾功能和氧化应激标记物的评估结果表明,牙周炎组的氧化应激水平异常,而肾功能水平出现异常,但无统计学意义。MitoSOX红染色结果显示,牙周炎组大鼠肾组织中ROS的含量明显高于对照组,RT-qPCR和免疫组化结果显示,牙周炎组大鼠肾组织中Bmal1、Nrf2和HO-1的表达水平呈下降趋势。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Impact of circadian clock protein Bmal1 on experimentally-induced periodontitis-associated renal injury.
OBJECTIVES To investigate the mechanism of circadian clock protein Bmal1 (Bmal1) on renal injury with chronic periodontitis, we established an experimental rat periodontitis model. METHODS Twelve male Wistar rats were randomly divided into control and periodontitis groups (n=6, each group). The first maxillary molars on both sides of the upper jaw of rats with periodontitis were ligated by using orthodontic ligature wires, whereas the control group received no intervention measures. After 8 weeks, clinical periodontal parameters, including probing depth, bleeding index, and tooth mobility, were evaluated in both groups. Micro-CT scanning and three-dimensional image reconstruction were performed on the maxillary bones of the rats for the assessment of alveolar bone resorption. Histopatholo-gical observations of periodontal and renal tissues were conducted using hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) staining. Renal function indicators, such as creatinine, albumin, and blood urea nitrogen levels, and oxidative stress markers, including superoxide dismutase, glutathione, and malondialdehyde levels, were measured using biochemical assay kits. MitoSOX red staining was used to detect reactive oxygen species (ROS) content in the kidneys. The gene and protein expression levels of Bmal1, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) in rat renal tissues were assessed using real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemical staining. RESULTS Micro-CT and HE staining results showed significant bone resorption and attachment loss in the maxillary first molar region of the periodontitis group. Histological examination through HE and PAS staining revealed substantial histopathological damage to the renal tissues of the rats in the periodontitis group. The findings of the assessment of renal function and oxidative stress markers indicated that the periodontitis group exhibited abnormal levels of oxidative stress, whereas the renal function levels showed abnormalities without statistical significance. MitoSOX Red staining results showed that the content of ROS in the renal tissue of the periodontitis group was significantly higher than that of the control group, and RT-qPCR and immunohistochemistry results showed that the expression levels of Bmal1, Nrf2, and HO-1 in the renal tissues of the rats in the periodontitis group showed a decreasing trend. CONCLUSIONS Circadian clock protein Bmal1 plays an important role in the oxidative damage process involved in the renal of rats with periodontitis.
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