地塞米松介导肠道炎症及肠黏膜屏障功能对斑马鱼脑损伤修复的抑制作用  

The inhibitory effects of dexamethasone on brain injury repair in zebrafish through intestinal inflammation and intestinal mucosal barrier function

作　　者：姚莉 张耀东 张熙佳 李欣欣 郭峻峰 何太平 崔春 尹恒[2,3] YAO Li;ZHANG Yaodong;ZHANG Xijia;LI Xinxin;GUO Junfeng;HE Taiping;CUI Chun;YIN Heng(Neurodegeneration and Neuroinjury Research Laboratory,Wuxi School of Medicine,Jiangnan University,Wuxi 214122,China;Wuxi TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Department of TCM Orthopedics,Wuxi 214071,China;Nanjing University of Chinese Medicine,Nanjing 210046,China)

机构地区：[1]江南大学无锡医学院神经退行和损伤研究室,江苏无锡214122 [2]南京中医药大学无锡附属医院骨伤科,江苏无锡214071 [3]南京中医药大学骨伤科研究所,江苏南京210046

出　　处：《延安大学学报（医学科学版）》2022年第3期7-13,F0002,共8页Journal of Yan'an University:Medical Science Edition

基　　金：国家自然科学基金(81973878);无锡市首届“双百”中青年医疗卫生拔尖人才项目(BJ2020063);江苏省大学生创新创业训练计划项目(202010295035Z)。

摘　　要：目的基于斑马鱼创伤性脑损伤(traumatic brain injury,TBI)后的可再生性修复功能,探究地塞米松(dexamethasone,Dex)处理对斑马鱼视顶盖损伤处细胞增殖的影响及损伤后恢复的作用及其机制。方法通过对斑马鱼进行地塞米松预处理(5 mg/L和15 mg/L)和视顶盖损伤,分别构建4组模型:空白组,损伤组,5 mg/L Dex+损伤组,15 mg/L Dex+损伤组。采用免疫荧光检测损伤处增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达;实时荧光定量PCR检测肠道中炎症相关分子(TNF-α、IL-6、TLR-2)及肠黏膜屏障相关分子(MUC2.1、DEFBL1、OCCLUDIN、TJP2α)mRNA表达。结果损伤组损伤侧视顶盖周围PCNA阳性细胞数在损伤后48 h(48 hours post injury,48 hpi)时较未损伤侧明显增多(P<0.01),而Dex(5 mg/L和15 mg/L)预处理+损伤组视顶盖细胞增殖则受到明显抑制。损伤组肠道中肿瘤坏死因子(tumor necrosis factor-α,TNF-α)、白介素-6(interleukin-6,IL-6)在48 hpi明显回复表达,而15 mg/L Dex+损伤组肠道炎症因子的表达则受到明显抑制。Toll样受体2(toll-like receptor 2,TLR-2)mRNA表达水平在损伤组48 hpi表达明显升高,而Dex处理显著抑制其表达。Dex抑制黏液素2.1(mucin 2.1,MUC2.1)在48 hpi的升高表达。β防御素样肽-1(defensin beta-like 1,DEFBL1)、紧密连接蛋白(occludin,OCCLUDIN)在12 hpi呈现被抑制表达。结论Dex可通过介导肠道炎症及肠黏膜屏障功能,抑制斑马鱼创伤性脑损伤后损伤部位的再生性细胞增殖过程,抑制损伤修复。Objective Based on the renewable repair function after traumatic brain injury(TBI)in zebrafish,this study explored the effects and mechanisms of dexamethasone(Dex)treatment on the proliferation of the optic tectum cells during the recovery post-injury.Methods Zebrafishes were treated with Dex pretreatment(5 mg/L and 15 mg/L)and optic tectum injury to construct 4 groups of models:blank group,injury group,5 mg/L Dex+injury group and 15 mg/L Dex+injury group.The expression levels of proliferating cell nuclear antigen(PCNA)were detected by immunofluorescence.The mRNA expression levels of intestinal inflammatory molecules(TNF-α,IL-6,TLR-2)and intestinal mucosal barrier molecules(MUC2.1,DEFBL1,OCCLUDIN,TJP2α)were detected by real-time quantitative PCR.Results The number of PCNA positive cells around the optic tectum of the injured side was significantly increased at 48 hpi compared with that of the uninjured side(P<0.01),while the proliferation of the optic tectum cells in the Dex(5 mg/L and 15 mg/L)pretreatment+injury group was significantly inhibited.The expression levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the intestine of the injury group were significantly restored at 48 hpi,while the expression levels of the inflammatory factors were significantly inhibited in the 15 mg/L Dex+injury group.Toll-like receptor 2(TLR-2)mRNA expression was significantly increased in the 48 hpi in the injury group,while Dex treatment significantly inhibited its expression.Dex inhibited the increased expression of mucin 2.1(mucin 2.1)at 48 hpi.Defensin beta-like 1(DEFBL1)and occludin(OCCLUDIN)were inhibited at 12 hpi.Conclusion Dex can inhibit the process of regenerative cell proliferationin the injury site and the repair of traumatic brain injury in zebrafish by mediating intestinal inflammation and intestinal mucosal barrier function.

关 键 词：创伤性脑损伤 地塞米松 肠道炎症 肠黏膜屏障 斑马鱼 

分 类 号：R742[医药卫生—神经病学与精神病学]