JNK/NF-κB/Caspase-3在小鼠实验性牙周炎诱导肾细胞凋亡中的作用研究  

Study on the role of c-Jun N-terminal Kinase/Nuclear Factor-κB/Caspase-3 in renal cell apoptosis induced by experimental periodontitis in mice

作　　者：傅若冰 邓愉 刘歆婵[2] 商雅琦 胡梦婷 车贞贞 于维先[1,3,4] FU Ruobing;DENG Yu;LIU Xinchan;SHANG Yaqi;HU Mengting;CHE Zhenzhen;YU Weixian(Department of Perio-dontics,Hospital of Stomatology,Jilin University,Changchun 130021,China;VIP Department,Hospital of Stomatology,Jilin University,Changchun 130021,China;Department of Geriatric Stomatology,Hospital of Stomatology,Jilin University,Changchun 130021,China;Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling,Changchun 130021,China)

机构地区：[1]吉林大学口腔医院牙周病科,吉林长春130021 [2]吉林大学口腔医院VIP综合科,吉林长春130021 [3]吉林大学口腔医院老年口腔科,吉林长春130021 [4]吉林省牙发育及颌骨重塑与再生重点实验室,吉林长春130021

出　　处：《口腔生物医学》2025年第1期26-33,共8页Oral Biomedicine

基　　金：吉林省科技厅国际合作项目(20220402069GH)。

摘　　要：目的:构建小鼠实验性牙周炎模型,探究c-Jun氨基末端激酶(JNK)/核因子-κB(NF-κB)/含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)在牙周炎诱导的肾功能异常中的作用。方法:选取12只SPF级雄性C57小鼠随机分成对照组、牙周炎组,后者上颌第二磨牙颈部结扎丝线构建牙周炎模型,4周后处死,显微CT(Micro-CT)评估牙槽骨吸收情况;苏木精-伊红(HE)和过碘酸雪夫(PAS)染色观察小鼠牙周及肾脏的病理变化;测定肾脏丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)及血清中肾功能指标肌酐(Cre)和白蛋白(Alb)的含量。线粒体超氧化物红色荧光探针(MitoSOX)染色、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)染色评估肾脏氧化应激及凋亡情况;实时荧光定量PCR分析小鼠肾组织中NF-κB、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、B细胞淋巴瘤-2基因(Bcl-2)及Bcl-2相关X蛋白(Bax)的表达水平。免疫组织化学染色测定肾脏JNK、磷酸化c-Jun氨基末端激酶(p-JNK)、NF-κB和Caspase-3的表达水平。结果:牙周炎组小鼠上颌第二磨牙牙龈红肿,Micro-CT见牙槽骨吸收,釉牙骨质界至牙槽嵴顶距离增加;牙周炎小鼠牙龈上皮钉突不规则伸长,伴牙槽嵴顶吸收,肾小球结构损伤及包曼氏囊腔扩张;牙周炎小鼠的肾小球基底膜基质增厚,肾小管刷状缘减少。牙周炎组的肾MDA、血清Cre和Alb升高(P<0.05),肾SOD和GSH下降(P<0.05);牙周炎小鼠肾脏活性氧(ROS)(P<0.05)及细胞凋亡数量均增多(P<0.05);牙周炎小鼠肾组织中IL-6、TNF-α、Bax、NF-κB的表达上升(P<0.05),Bcl-2表达下降(P<0.05),JNK、p-JNK、NF-κB和Caspase-3的表达显著增加(P<0.05)。结论:JNK/NF-κB/Caspase-3可能通过影响肾细胞凋亡,在牙周炎诱导的肾功能异常中发挥重要作用。Objective:To investigate the role of c-Jun N-terminal kinase(JNK)/nuclear factor-κB(NF-κB)/Caspase-3 in kidney dysfunction related to periodontitis by establishing a mouse model of experimental periodontitis.Methods:Twelve SPF-grade male C57 mice were randomly divided into a control group and a periodontitis group.Periodontitis was induced by ligating silk around the necks of the bilateral maxillary second molars.Four weeks later,the mice were sacrificed,and the three-dimensional structure of the alveolar bone was reconstructed using Micro-CT to analyze bone resorption.Histological changes in periodontal and kidney tissues were observed using hematoxylin-eosin(HE) and periodic acid-schiff(PAS) staining.Biochemical kits were used to detect kidney oxidative stress biomarkers malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),and kidney function indicators creatinine(Cre) and albumin(Alb).Mitochondrial superoxide indicator(MitoSOX) staining was employed to assess ROS levels in kidney tissues,while terminal deoxynucleotidyl transferase-mediated dUTPnick end labeling(TUNEL) staining evaluated renal cell apoptosis.Quantitative real-time polymerase chain reaction(qRT-PCR) was used to measure expression levels of NF-κB,interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),B-cell lymphoma 2(Bcl-2),and B-cell lymphoma-2-associated X protein(Bax) in kidney tissues.Immunohistochemistry was performed to detect phosphorylated JNK(p-JNK),JNK,NF-κB,and Caspase-3 expressions in kidney tissues.Results:Compared to the control group,mice in the periodontitis group exhibited gingival hyperemia and swelling around the maxillary second molars.Micro-CT analysis revealed significant alveolar bone resorption in the periodontitis group,with an increased distance from the enamel-dentin junction to the alveolar ridge crest.Irregular elongation of the gingival epithelial rete pegs,accompanied by resorption at the alveolar ridge crest,damage to the glomerular structure,and expansion of Bowman's space were showed.The thickening

关 键 词：牙周炎 肾功能异常 C-JUN氨基末端激酶 核因子-κB 含半胱氨酸的天冬氨酸蛋白水解酶-3 凋亡 

分 类 号：R781.42[医药卫生—口腔医学]