摘要
目的 探讨口腔牙龈卟啉单胞菌(Pg)能否通过调控TLR4/NF-κB信号通路影响围手术期认知功能障碍(PND)的发生。方法 细胞实验取对数生长期的BV-2细胞随机分为两组:对照组(C组)、LPS-Pg处理组(P组),采用Western blot实验检测细胞中信号蛋白(TLR4、NF-κB p65、NLRP3、TNF-α)的表达,通过免疫荧光实验检测NF-κB p65在细胞核内外的表达变化。动物实验将C57BL/6小鼠随机分为6组:对照组(C组)、手术组(S组)、Pg感染组(P组)、Pg感染+手术组(PS组)、TLR4抑制剂组(T组)、Pg感染+手术+TLR4抑制剂组(PST组),对各组进行相应的处理后,应用水迷宫测试观察Pg感染对小鼠术后认知行为的影响,应用组织免疫化学实验观察Pg感染对小鼠海马组织中小胶质细胞活化的影响。结果 细胞实验结果显示,与C组比较,Western blot检测发现P组的TLR4、NLRP3、TNF-α表达量明显增高(均P<0.05),NF-κB在核内的表达量增加(P<0.05)、核外表达量减少(P<0.05),免疫荧光实验显示P组NF-κB表达量增多且向核内转移。动物实验结果显示,水迷宫实验结果显示,各组小鼠在术前训练阶段的表现并无显著差异,其逃避潜伏期与平均游泳速度均处于相近水平(均P>0.05)。术后测试阶段的结果显示,相较于C组,S组和PS组小鼠在目标象限的停留时间更短,穿越平台的次数也较少(均P<0.05)。与PS组比较,PST组小鼠在目标象限的停留时间较长,且穿越平台的次数也较多(均P<0.05)。对海马组织进行免疫组化分析提示PS组中的阳性细胞数量显著多于C组和PST组,这些阳性细胞的胞体增大,突起变短,且小胶质细胞呈现激活状态。结论 Pg感染诱发小鼠围手术期中枢神经系统炎症反应增强,引起小鼠PND的发生,可能与激活TLR4/NF-κB信号通路有关。
Objective To investigate whether Porphyromonas gingivalis(Pg)in the oral cavity can influence the occurrence of perioperative neurocognitive disorder(PND)by modulating the TLR4/NF-κB signaling pathway.Methods In cell experi-ment,BV-2 cells in the logarithmic growth phase were randomly divided into two groups:the control group(Group C)and the LPS-Pg-treated group(Group P).Western blot was used to detect the expression levels of signaling proteins(TLR4,NF-κB p65,NLRP3,TNF-α)in cellular proteins.Immunofluorescence assay was employed to observe the changes in NF-κB p65 expres-sions inside and outside the nucleus.In animal experiments,C57BL/6 mice were randomly divided into six groups:the control group(Group C),the surgery group(Group S),the Pg infection group(Group P),the Pg infection+surgery group(Group PS),the TLR4 inhibitor group(Group T),and the Pg infection+surgery+TLR4 inhibitor group(Group PST).After corresponding treatments,the Morris water maze(MWM)test was applied to observe the effects of Pg infection on postoperative cognitive be-havior in mice.Immunohistochemical analysis was used to assess the impact of Pg infection on the activation of microglia in the hippocampal tissue of mice.Results In cell experiment,compared with Group C,expression levels of TLR4,NLRP3,and TNF-αincreased significantly in Group P(all P<0.05).The expression of NF-κB p65 in the nucleus increased(P<0.05),while its expression outside the nucleus decreased(P<0.05).Immunofluorescence results showed increased NF-κB expression and its translocation into the nucleus in Group P.As for animal experiment,no significant difference was observed in the preoperative training phase among the groups in Morris water maze experiment,with similar escape latencies and average swimming speeds(all P>0.05).Therefore,there was no notable differences in navigation and swimming abilities among the groups before the ex-periment.In the postoperative testing phase,compared with Group C,the target quadrant time and platform-crossing times were less in mice of Group S and Group PS(all P<0.05).Compared with Group PS,the target quadrant time and platform-crossing times were elevated in mice of Group PST(both P<0.05).Compared to Group C and Group PST,number of positive cells in hippocampal tissue was higher in Group PS.These positive cells exhibited enlarged somas,shortened processes,and an activated state of microglia.Conclusion Pg infection induces enhanced central nervous system inflammation during the perioperative peri-od in mice,leading to the occurrence of PND,possibly through the activation of the TLR4/NF-κB signaling pathway.
作者
谢小娟
张爽
柏效治
郝雪琴
吴瑶瑶
郑鹏
Xie Xiaojuan;Zhang Shuang;Bai Xiaozhi(Department of Anesthesiology,College of Clinical Medicine,the Fist Af filiated Hospital of Henan University of Science and Technology,Luoyang 471003,China)
出处
《华中科技大学学报(医学版)》
北大核心
2025年第4期483-490,共8页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
河南省医学科技攻关计划项目(No.LHGJ20220690)
河南省高等学校重点科研项目(No.22A320032)。
