MLKL在小鼠皮肤伤口愈合过程中的作用  

The role of MLKL in mediating skin wound healing in Mice

作　　者：包文慧 赵佳敏 巩志国 白云洁 于琢雅 刘博[1] 毛伟[1] 张双翼[1] BAO Wenhui;ZHAO Jiamin;GONG Zhiguo;BAI Yunjie;YU Zhuoya;LIU Bo;MAO Wei;ZHANG Shuangyi(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018

出　　处：《中国农业大学学报》2024年第10期230-238,共9页Journal of China Agricultural University

基　　金：国家自然科学基金项目(32202879);内蒙古农业大学优秀博士人才引进科研启动项目(NDYB2021-19);内蒙古自治区高等学校科学研究重点项目(NJZZ23035)。

摘　　要：为探究MLKL对肌成纤维细胞和巨噬细胞间相互作用过程中的调控作用及对小鼠皮肤伤口愈合的影响,分别构建野生型C57BL/6J小鼠和MLKL基因敲除(MLKL^(-/-))小鼠背部创伤模型,分析皮肤伤口愈合情况。体外培养野生型C57BL/6J和MLKL^(-/-)小鼠肌成纤维细胞及骨髓源巨噬细胞,使用小鼠耳源肌成纤维细胞培养上清液(Myofibroblast conditioned medium,MFbCM)处理野生型C57BL/6J小鼠M1和M2型巨噬细胞,检测M1型巨噬细胞IL-6、TNF-α及NO的分泌量。检测M2型巨噬细胞精氨酸酶(Arginase)及几丁质酶3样分子(Ym1)的表达量。结果显示,1)相比于野生型C57BL/6J小鼠,MLKL^(-/-)小鼠伤口收缩更为缓慢。在损伤后第5、7和10天,野生型C57BL/6J小鼠创面收缩率显著高于MLKL^(-/-)小鼠(P<0.001)。2)MLKL缺失显著降低伤口表面Ym1和Arginase的表达,在损伤后第5、7和10天差异显著(P<0.001)。此外,MLKL的缺失影响了MFbCM上调M2型巨噬细胞Ym1和Arginase表达的能力。表明MLKL可能参与调控MFbCM诱导的M2型巨噬细胞分泌活动。3)MFbCM可促进M1型巨噬细胞IL-6、TNF-α和NO的表达,且依赖于MLKL的存在,表明MLKL可能参与调控MFbCM诱导的M1型巨噬细胞分泌活动。综上,本研究确定了MLKL参与巨噬细胞和肌成纤维细胞间相互作用,这种相互作用在伤口愈合的每个阶段都是必不可少的。因此,该研究将有助于更好地理解MLKL在创伤愈合中的作用,为创面愈合提供新的治疗策略。To explore the regulatory effect of MLKL on the interaction between myofibroblasts and macrophages and its effect on skin wound healing in mice,this study constructed back trauma models of wild-type C57BL/6J mice and MLKL knockout(MLKL^(-/-))mice to analyze skin wound healing.Myofibroblasts and bone marrow-derived macrophages of Wild-type C57BL/6J mice and MLKL^(-/-)mice were cultured in vitro.M1 and M2 macrophages of wild-type C57BL/6J mice were treated with mouse ear skin myofibroblast conditioned medium(MFbCM),and the secretion of IL-6,TNF-αand NO in M1 macrophages were detected.The expression of Arginase and chitinase 3-like molecule(Ym1)in M2 macrophages was detected.The results showed that:1)Compared with wild-type C57BL/6J mice,MLKL^(-/-)mice had slower wound contraction.On the 5th,7th and 10th day after injury,the wound contraction rate of wild-type C57BL/6J mice was significantly higher than that of MLKL^(-/-)mice(P<0.001).2)MLKL deficiency significantly reduced the expression of Ym1 and Arginase on the wound surface,which were more significant on the 5th,7th and 10th day after injury(P<0.001).In addition,the deletion of MLKL affected the ability of MFbCM to up-regulate the expression of Ym1 and Arginase in M2 macrophages,suggesting that MLKL may be involved in the regulation of MFbCM-induced M2 macrophage secretion.3)MFbCM can promote the expression of IL-6,TNF-αand NO in M1 macrophages,which was dependent on the presence of MLKL.It was suggested that MLKL may be involved in the regulation of MFbCM-induced M1 macrophage secretion.In summary,this study identified that MLKL is involved in the interaction between macrophages and myofibroblasts,which is essential at every stage of wound healing.Therefore,this study will help to better understand the role of MLKL in wound healing and provide new therapeutic strategies for wound healing.

关 键 词：MLKL 创伤愈合 巨噬细胞 肌成纤维细胞 

分 类 号：S859.7[农业科学—临床兽医学]