CaMKIV信号缺失促进肌损伤炎症并影响肌再生

doi:10.13418/j.issn.1001-165x.2023.6.11

中国临床解剖学杂志 ›› 2023, Vol. 41 ›› Issue (6): 692-697.doi: 10.13418/j.issn.1001-165x.2023.6.11

CaMKIV信号缺失促进肌损伤炎症并影响肌再生

王涵¹，厉洋洋¹，菅晓婷¹，黄静雯¹，蓝海强¹，胡稷杰²*，廖华¹*

1.广东省组织构建与检测重点实验室，南方医科大学基础医学院解剖教研室，广州 510515；
2.南方医科大学南方医院创伤骨科，广州 510515

收稿日期:2023-04-06 出版日期:2023-11-25 发布日期:2023-12-26
通讯作者: 胡稷杰，副主任医师，E-mail: zhuchunmin@lzu.edu.cn；廖华，教授，E-mail: hua-liao@163.com
作者简介:王涵（1998-），男，湖南人，硕士研究生，研究方向：骨骼肌损伤，E-mail:wh2211@163.com
基金资助:
国家自然科学基金面上项目（32071181）；广州市科技计划项目（202002030497）；广东省自然科学基金面上项目（2023A1515012191）；国家重点研发计划（2022YFF1202603）

Loss of CaMKIV signaling aggravates muscle inflammation and affects muscle regeneration

Wang Han¹, Li Yangyang¹, Jian Xiaoting¹, Huang Jingwen¹, Lan Haiqiang¹, Hu Jijie²*, Liao Hua¹*

1. Guangdong Provincial Key Laboratory of Tissue Construction and Testing, Department of Anatomy, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China; 2. Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China

Received:2023-04-06 Online:2023-11-25 Published:2023-12-26

摘要/Abstract

摘要： 目的探究CaMKIV信号对肌损伤炎症反应及肌修复的调控作用。方法采用CRISPR/Cas9技术构建CaMKIV基因缺失（CaMKIV-/-）小鼠，PCR表型鉴定。心脏毒素（Cardiotoxin ，CTX)诱导野生B6鼠及CaMKIV-/-鼠急性肌损伤。HE染色观察损伤肌内炎症程度及肌修复过程。Dystrophy、F4/80免疫荧光双标，观察损伤肌内巨噬细胞渗出差异。F4/80、Ly-6C、CD206、Ki67染色及流式检测，分析损伤肌内M1、M2巨噬细胞的数量和比例，以及M1、M2巨噬细胞的局部增殖能力。结果成功构建并繁殖CaMKIV-/-鼠。HE及荧光染色观察证实小鼠体内CaMKⅣ信号缺失可致肌内炎症加剧、肌修复延迟。CaMKⅣ-/-小鼠损伤肌内再生的中央核肌纤维（Dystrophin+）数量低于WT鼠，巨噬细胞（F4/80+）数量显著高于WT鼠，以M1细胞（F4/80+Ly-6C+）为主，M2细胞（F4/80+CD206+）比例低于WT鼠。CaMKⅣ-/-鼠损伤肌内增殖巨噬细胞（F4/80+Ki67+）数量显著高于对照鼠，但以增殖的M1细胞为主(Ly6C+Ki67+)，M2细胞(CD206+Ki67+)的增殖与对照鼠无显著差异。结论 CaMKⅣ信号缺失可致肌损伤炎症加剧、促进损伤肌内M1巨噬细胞的渗出及增殖，延误肌修复。

关键词: 骨骼肌损伤, , , 钙调蛋白依赖性蛋白激酶4, , , 巨噬细胞, , , 炎症反应, , , , 增殖

Abstract: Objective To explore the effects of CaMKIV signaling on muscle inflammatory response and muscle repair post-injury. Methods CaMKIV gene deletion (CaMKIV-/-) mice were constructed by using CRISPR/Cas9 technology and identified by PCR. Cardiotoxin (CTX) induced skeletal muscle injury was performed in wild B6 and CaMKIV-/- mice. HE staining was used to observe the inflammation and repair in injured muscles. Dystrophy immunofluorescence staining was used to identify the regenerating myofibers while F4/80 staining to present infiltrated macrophage in inflamed muscle. F4/80, Ly-6C, CD206, Ki67 staining and flow cytometry skill were used to analyze the number and proportion of M1 (F4/80+Ly6C+) and M2 ((F4/80+CD206+) macrophages in the injured muscle, as well proliferation capacity of macrophages (F4/80+Ki67+). Results CaMKIV-/- mice were successfully constructed and bred. HE and fluorescence staining showed that, the absence of CaMKⅣ signaling in mice resulted in increasing intramuscular inflammation and delaying muscle repair. The number of the regenerating central nuclear muscle fibers（Dystrophin+） in CaMKⅣ-/- mice was lower than that in WT mice. The number of macrophages (F4/80+) in inflamed muscle from CaMKⅣ-/- mice was significantly higher than that of WT mice, which was dominant by M1 cells, comparing with that of WT mice. In inflamed muscle of CaMKⅣ-/- mice, the number and proportion of proliferating macrophages (F4/80+Ki67+) was significantly higher than that of control mice, but the proliferating macrophages were mainly M1 cells (Ly6C+Ki67+). Conclusions The loss of CaMKⅣ signaling leads to the aggravation of muscle inflammation, promotes the exudation and proliferation of M1 macrophages in inflamed muscle, and delays muscle repair after injury.

Key words: Muscle injury, , , CaMKⅣ, , , Macrophage, , , Inflammation, , , Proliferation

中图分类号:

R392.31

王涵, 厉洋洋, 菅晓婷, 黄静雯, 蓝海强, 胡稷杰, 廖华. CaMKIV信号缺失促进肌损伤炎症并影响肌再生[J]. 中国临床解剖学杂志, 2023, 41(6): 692-697.

Wang Han, Li Yangyang, Jian Xiaoting, Huang Jingwen, Lan Haiqiang, Hu Jijie, Liao Hua. Loss of CaMKIV signaling aggravates muscle inflammation and affects muscle regeneration[J]. Chinese Journal of Clinical Anatomy, 2023, 41(6): 692-697.

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CaMKIV信号缺失促进肌损伤炎症并影响肌再生

Loss of CaMKIV signaling aggravates muscle inflammation and affects muscle regeneration

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