脑和肌肉芳香烃受体核转运样蛋白1在大鼠牙周炎诱导肝损伤模型中的作用研究  

Role of brain and muscle ARNT-like protein 1 in the rat periodontitis-induced liver injury model

作　　者：刘笑梦 曹牛奔 邓愉 侯玉帛 刘歆婵[2] 马浩楠 于维先[3] Liu Xiaomeng;Cao Niuben;Deng Yu;Hou Yubo;Liu Xinchan;Ma Haonan;Yu Weixian(Department of Periodontics,Hospital of Stomatology,Jilin University,Changchun 130021,China;Department of Dental Implantology,Hospital of Stomatology,Jilin University,Changchun 130021,China;Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling,Hospital of Stomatology,Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学口腔医院牙周病科,长春130021 [2]吉林大学口腔医院种植科,长春130021 [3]吉林大学口腔医院吉林省牙发育及颌骨重塑与再生重点实验室,长春130021

出　　处：《中华口腔医学杂志》2022年第10期1048-1056,共9页Chinese Journal of Stomatology

基　　金：吉林省财政厅科技项目(JCSZ2021893-22);吉林省科技发展计划(20190201058JC)。

摘　　要：目的探讨脑和肌肉芳香烃受体核转运样蛋白1(brain and muscle ARNT-like protein 1,BMAL1)在大鼠牙周炎诱导肝损伤模型中的作用。方法根据随机数字表法将12只Wistar雄性大鼠随机分为对照组和牙周炎组,每组6只。对照组大鼠不做处理。牙周炎组大鼠通过结扎双侧上颌第一磨牙颈部建立牙周炎模型。建模8周后检测两组大鼠牙周临床指标并处死。显微CT(micro-CT)扫描大鼠上颌骨并分析牙槽骨吸收情况。HE及油红O染色分析两组大鼠牙周组织和肝组织的病理变化。生化试剂盒检测血清中谷草转氨酶(glutamic-oxaloacetic transaminase,GOT)、谷丙转氨酶(glutamic-pyruvic transaminase,GPT)、总胆固醇(total cholesterol,TC)和甘油三酯(triglyceride,TG)等肝功能相关指标。实时荧光定量PCR(real time fluorescent quantitative-PCR,qRT-PCR)、免疫组化和蛋白质印迹法检测肝组织中BMAL1、核因子-κB(nuclear factor kappa-B,NF-κB)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)基因及蛋白的表达水平。原位末端转移酶标记(terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling,TUNEL)试剂盒染色法检测肝组织中的细胞凋亡。结果上颌第一磨牙HE染色和micro-CT结果显示,牙周炎组大鼠牙槽骨吸收明显。肝组织病理学结果显示,与对照组相比,牙周炎组肝组织内炎症细胞浸润、肝索结构紊乱且肝细胞中可见大量脂滴形成。大鼠血清生化检测结果显示,牙周炎组大鼠血清中GOT[(62.77±2.59)U/L]、GPT[(47.54±1.04)U/L]、TC[(3.19±0.23)mmol/L]和TG[(1.11±0.09)mmol/L]均较对照组GOT[(38.66±2.47)U/L]、GPT[(31.48±1.57)U/L]、TC[(1.60±0.05)mmol/L]和TG[(0.61±0.09)mmol/L]含量显著升高(P=0.003,P=0.001,P=0.002,P=0.038)。qRT-PCR结果显示,牙周炎组大鼠肝组织中BMAL1 mRNA的表达[(0.60±0.04)%]较对照组[(1.01±0.07)%]显著下调(t=4.80,P=0.009),NF-κB、TNF-αmRNA的表达[(1.62±0.12)%、(2.69±0.16)%]均较对照组[(1.00±Objective Brain and muscle ARNT-like protein 1(BMAL1)is a core component of hepatocyte molecular clock and plays an important role in the regulation of other related rhythmic genes in the body through a transcriptional-translational feedback loop in molecular circadian oscillations.Therefore,the aim of this study was to investigate the role of BMAL1 in the rat periodontitis-induced liver injury.Methods Twelve male Wistar rats were divided into the control group and the periodontitis group according to the random number table method.The rats in the control group were untreated.The periodontitis models were established by ligating the necks of the bilateral maxillary first molars in the periodontitis group rats.After 8 weeks,periodontal clinical indexes of rats in both groups were examined and executed.Micro-CT scans of the maxilla were performed and levels of the alveolar bone resorption were analyzed.Pathological changes in periodontal and liver tissue of rats in two groups were detected by HE and oil red O staining.Biochemical kits were used to detect glutamic-oxaloacetic transaminase(GOT),glutamic-pyruvic transaminase(GPT),total cholesterol(TC)and triglycerides(TG)in serum.The gene and protein expression levels of BMAL1,nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)in liver tissue were measured by real time fluorescent quantitative-PCR(qRT-PCR),immunohistochemistry(IHC)and Western blotting(WB)assays.Apoptosis was detected in liver tissues by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL)kit staining.Results The results of HE staining of maxillary first molars and micro-CT results of maxillary bones showed that alveolar bone resorption was significant in the periodontitis group of rats.The liver histopathology results showed infiltrated inflammatory cells in the liver tissue,disorganized liver cords and a large number of lipid droplets formed in the hepatocytes of the periodontitis group compared with the control group.The results of serum biochemical assa

关 键 词：牙周炎 芳香烃受体核转位子 肝损伤 动物 实验 脑和肌肉芳香烃受体核转运样蛋白1 大鼠 Wistar 

分 类 号：R-332[医药卫生] R575R781.42