Department of Internal Medicine, Section on Pulmonary, Critical Care, Allergy, and Immunologic Diseases, Wake Forest School of Medicine, Winston-Salem, NC, United States.
Department of Internal Medicine, Section on Pulmonary, Critical Care, Allergy, and Immunologic Diseases, Wake Forest School of Medicine, Winston-Salem, NC, United States.
J Immunol Methods. 2022 Oct;509:113329. doi: 10.1016/j.jim.2022.113329. Epub 2022 Aug 3.
Macrophages are important mediators of skeletal muscle function in both healthy and diseased states. In vivo specific depletion of macrophages provides an experimental method to understand physiological and pathophysiological effects of macrophages. Systemic depletion of macrophages can deplete skeletal muscle macrophages but also alters systemic inflammatory responses and metabolism, which confounds the muscle specific effects of macrophage depletion. The primary aim of this manuscript is to evaluate two methods of murine intramuscular macrophage depletion in an acute lung injury-associated indirect skeletal muscle wasting mouse model. Adult C57BL/6 (WT) and Macrophage Fas-Induced Apoptosis (MaFIA, C57BL/6-Tg) mice received clodronate liposomes or the dimerization drug AP20187 through intramuscular injection of the tibialis anterior muscle compartment, respectively. Vehicle control was injected in the contralateral muscle. We demonstrate intramuscular AP20187 in the MaFIA mouse depletes macrophages but causes an infiltration of CD45 intermediate neutrophils. In contrast, intramuscular clodronate liposomes successfully depletes macrophages without an associated increase in CD45 intermediate cells. In conclusion, intramuscular clodronate is effective for selective depletion of muscle macrophages without eliciting acute inflammation seen with AP20187 in MaFIA mice. This technique is an important tool to study the functional roles of macrophages in skeletal muscle.
巨噬细胞是健康和患病状态下骨骼肌功能的重要介质。体内特异性耗尽巨噬细胞为理解巨噬细胞的生理和病理生理效应提供了一种实验方法。系统性耗尽巨噬细胞可以耗尽骨骼肌巨噬细胞,但也会改变全身炎症反应和代谢,从而混淆巨噬细胞耗竭对肌肉的特异性影响。本文的主要目的是在急性肺损伤相关间接性骨骼肌消耗小鼠模型中评估两种小鼠肌肉内巨噬细胞耗竭方法。成年 C57BL/6(WT)和巨噬细胞 Fas 诱导凋亡(MaFIA,C57BL/6-Tg)小鼠分别通过胫骨前肌间隔肌内注射氯膦酸盐脂质体或二聚化药物 AP20187 接受治疗。对照侧肌肉注射载体对照。我们证明了 MaFIA 小鼠肌肉内的 AP20187 可以耗尽巨噬细胞,但会引起 CD45 中间中性粒细胞的浸润。相比之下,肌肉内氯膦酸盐脂质体成功耗尽了巨噬细胞,而没有伴随 MaFIA 小鼠中看到的 AP20187 引起的 CD45 中间细胞增加。总之,肌肉内氯膦酸盐有效地选择性耗尽肌肉巨噬细胞,而不会引起 MaFIA 小鼠中 AP20187 引起的急性炎症。这项技术是研究巨噬细胞在骨骼肌中功能作用的重要工具。