P2X受体在牙周炎模型大鼠牙龈组织中的表达变化

Expression Change of P2X Receptors in Gingival Tissues of Experimental Periodontitis Rats

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摘要目的探究P2X受体在牙周炎模型大鼠牙龈组织中的表达及变化,鉴定牙周炎发生发展的相关P2X受体亚型。方法将40只雄性SD大鼠随机分为4组(均n=10):对照组、2周牙周炎组(2WP组)、4周牙周炎组(4WP组)、6周牙周炎组(6WP组)。采用0.2 mm的正畸结扎丝结扎上颌双侧第一磨牙法建立实验性牙周炎大鼠模型,取分别结扎2、4、6周的2WP组、4WP组、6WP组以及对照组牙龈组织进行研究。通过牙龈指数(GI)、颌骨Micro-CT扫描与亚甲基蓝染色的方法评估各组大鼠牙周情况。采用实时荧光定量PCR检测各组大鼠牙龈组织中P2X1—7亚型受体的mRNA表达水平。采用蛋白免疫印迹方法与免疫组织化学检测各组大鼠牙龈组织中P2X7受体的蛋白表达水平。采用ELISA法检测各组血清中TNF-α与IL-1β的水平变化。结果随着牙周结扎时间的延长,大鼠炎症程度增加,牙槽骨的吸收逐渐增多(P<0.01)。6WP组牙龈组织中P2X1、P2X2、P2X5、P2X6、P2X7受体mRNA表达和正常大鼠相比显著升高(P<0.05),P2X4受体mRNA表达明显下降(P<0.05),P2X3受体mRNA几乎不表达。与对照组相比,牙周炎模型各组牙龈组织中P2X7受体的蛋白含量显著上升(P<0.05),且P2X7受体主要表达于牙龈组织的口腔上皮。与对照组比较,各牙周炎模型组血清中TNF-α与IL-1β水平均明显升高(P<0.01),且TNF-α与IL-1β水平随着结扎时间的延长而增加(P<0.05)。结论牙周炎模型大鼠牙龈组织中P2X1—7亚型受体mRNA表达水平变化明显,其中P2X7受体表达水平升高最为显著,P2X7受体可能在牙周炎的进展中发挥一定作用。 Objective To investigate the expression and changes of P2X receptors in gingival tissues of experimental periodontitis rats and to find out the P2X receptor subtypes related to the development of periodontitis.Methods 40 male SD rats were randomly divided into 4 groups(n=10 for each group):control group,2-week periodontitis group(2WP group),4-week periodontitis group(4WP group),and 6-week periodontitis group(6WP group).The experimental periodontitis rat model was established by ligating the bilateral maxillary first molars with 0.2 mm orthodontic ligation wire,and the gingival tissues of the rats in each group were collected for observation at 2,4,and 6 weeks after periodontal ligation.The periodontal conditions of rats in each group were evaluated using gingival index(GI),jaw Micro-CT scanning,and methylene blue staining.Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of P2X1-7 subtype receptors in gingival tissues of rats in each group;western blot and immunohistochemistry were employed to assess the protein expression levels of P2X7 receptors in gingival tissues of rats in each group;the changes in the concentrations of TNF-αand IL-1βin serum in each group were measured by ELISA.Results With the extension of periodontal ligation time,the degree of inflammation increased and the absorption of alveolar bone gradually increased(P<0.01).The mRNA expression levels of P2X1,P2X2,P2X5,P2X6 and P2X7 receptors in gingival tissues of the 6WP group were significantly increased compared to those of normal rats(P<0.05),whereas the mRNA expression of P2X4 receptors was markedly reduced(P<0.05)and the mRNA of P2X3 receptors was almost not detected.Compared to the control group,the protein contents of P2X7 receptors in gingival tissues of each periodontitis model group were significantly increased(P<0.05),and P2X7 receptors were mainly expressed in the oral epithelium of gingival tissues.Compared to the control group,the serum concentrations of TNF-αand IL-1βin each periodontitis model

作者左月周敏刘丽娟周昱涵韩伟李鑫哲谢玉婷邱妍婷高云熊伟 ZUO Yue;ZHOU Min;LIU Li-juan;ZHOU Yu-han;HAN Wei;LI Xin-zhe;XIE Yu-ting;QIU Yan-ting;GAO Yun;XIONG Wei(Department of Preventive Health Care,the Affiliated Stomatological Hospital,Nanchang University;Department of Physiology,School of Basic Medical Sciences,Nanchang University;Jiangxi Key Laboratory of Oral Biomedical Sciences,Nanchang 330006,China)

机构地区南昌大学附属口腔医院预防保健科南昌大学基础医学院生理教研室江西省口腔生物医学重点实验室

出处《南昌大学学报（医学版）》 2024年第6期1-7,41,共8页 Journal of Nanchang University：Medical Sciences

基金国家自然科学基金(81860199,82360199)。