摘要
目的:探讨嗅细胞死亡及其对损伤后反应性细胞增殖的影响。方法:采用C57BL/6野生型小鼠、受体相互作用丝苏氨酸蛋白激酶3(RIPK3)基因敲除小鼠、混合谱系激酶结构域样假激酶(MLKL)基因敲除小鼠和甲巯咪唑制备嗅上皮损伤模型;采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)染色、活体碘化丙啶(PI)标记法观察细胞死亡。采用Western Blot和免疫荧光染色检测程序性细胞坏死关键蛋白分子RIPK3、MLKL及磷酸化(p-MLKL)水平变化。采用BrdU掺入法研究细胞增殖。结果:嗅上皮损伤后3 d,嗅细胞主要发生坏死,少量发生凋亡。嗅上皮的RIPK3上调且MLKL磷酸化水平增加。BrdU掺入实验表明:程序性坏死关键分子RIPK3或MLKL基因敲除可促进小鼠嗅细胞反应性增殖。结论:嗅上皮损伤后早期,主要发生程序性坏死。阻断程序性细胞坏死可促进嗅细胞前体细胞的反应性增殖。
Objective:To explore the effects of death of olfactory neural cells on the injury-induced cell proliferation in olfactory epithelium(OE).Methods:Wild type(WT)C57BL/6 mice,receptor-interacting serine/threonine-protein kinase 3(RIPK3)mutant mice,mixed lineage kinase domain like pseudokinase(MLKL)mutant mice and thiamazole were adopted to make olfactory injury.Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining and in vivo propidium iodide(PI)labeling were used for detecting cell death.Western-blotting and immunohistochemistry were used for evaluating the expression of necroptotic markers.BrdU labelling was used to assess cell proliferation.Results:TUNEL-and PI-staining showed that most olfactory cells died through necrosis while few through apoptosis at 3 d post injury.Immunohistochemistry and Western Blot showed that necroptotic markers RIPK3 and phosphorylated MLKL were up-regulated in OE at 3 d post injury.The BrdU-incorporation assay showed that depleting RIPK3 or MLKL significantly increased the number of BrdU-positive cells in OE.Conclusion:Olfactory cells die mainly through necroptosis at the early phase of injury.Blocking necroptosis could increase the responsive proliferation of olfactory cells,thus facilitating neuronal regeneration in OE.
作者
符帆帆
陈玉洁
郑威安
孟欣宇
杨彦玲
武胜昔
王亚周
赵琳
FU Fanfan;CHEN Yujie;ZHENG Wei'an;MENG Xinyu;YANG Yanling;WU Shengxi;WANG Yazhou;ZHAO Lin(Department of Anatomy,School of Medicine,Yan'an University,Yan'an 716099;Department of Neurobiology,School of Basic Medicine,Air Force Medical University,Xi'an 710032,China)
出处
《神经解剖学杂志》
CAS
CSCD
2023年第1期39-46,共8页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金(31970907)。
关键词
嗅上皮
程序性细胞坏死
增殖
小鼠
olfactory epithelium
programmed cell necroptosis
proliferation
mouse
