皮下注射巨噬细胞集落刺激因子诱导机械痛超敏的性别二态性  

Sexual Dimorphism in Mechanical Hypersensitivity Induced by Subcutaneous Injection of M-CSF

作　　者：彭湘格 林震嘉 张辉[2] 冷世泽 唐源 郑钰凡 谈智[1] 周利君[1] PENG Xiang-ge;LIN Zhen-jia;ZHANG Hui;LENG Shi-ze;TANG Yuan;ZHENG Yu-fan;TAN Zhi;ZHOU Li-jun(Department of Physiology,Zhongshan School of Medicine,Sun Yat-sen University/Center for Pain Research,Sun Yat-sen University,Guangzhou 510080,China;Department of Anesthesiology,Guangdong Second People′s Hospital,Guangzhou 510317,China)

机构地区：[1]中山大学中山医学院生理教研室/中山大学疼痛研究中心,广东广州510080 [2]广东省第二人民医院麻醉科,广东广州510317

出　　处：《中山大学学报（医学科学版）》2023年第3期395-402,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金(2021A1515011201,2021A1515011742,2020A1515010204);广东省第二人民医院2019年医院引进人才科研启动基金(YY2019-003);广州市科技技术局市校(院)联合资助项目(SL2023A03J00808)。

摘　　要：【目的】通过正常小鼠皮下注射巨噬细胞集落刺激因子(M-CSF)来研究其对疼痛行为的影响是否存在性别差异,并初步探讨其机制。【方法】将30只10周龄C57 BL/6J小鼠随机分为三组(每组10只,雌雄各半):牛血清白蛋白对照组(BSA,0.3μg)、低剂量M-CSF组(L M-CSF,0.075μg)、高剂量M-CSF组(H M-CSF,0.3μg)。分别于左大腿内侧皮下注射50μL BSA或M-CSF,每日1次,连续3 d。给药前后采用von-Frey机械敏感性测试检测各组小鼠的机械撤足阈值(PWT)。再用免疫荧光染色法观察皮肤离子钙结合衔接蛋白1(Iba1),L5-L6 DRG及腰段脊髓背角中降钙素基因相关肽(CGRP)、磷酸化细胞外信号调节激酶(p-ERK)的表达变化。【结果】在雌性小鼠中,只有高剂量M-CSF引起显著的机械痛超敏,而在雄性小鼠中两个剂量都引起明显痛觉异常。机制上,高剂量M-CSF诱导雌性小鼠皮下巨噬细胞大量聚集(Iba1标识);雄性小鼠巨噬细胞呈浓度依赖性增多,但没有雌性小鼠明显。在DRGs中,p-ERK与CGRP表达增多也存在类似的性别差异。值得注意的是,CGRP在雄性小鼠DRG的神经纤维中显著升高。相应地,脊髓背角浅层的p-ERK与CGRP+末梢在M-CSF处理后显著增多,且雄性小鼠多于雌性小鼠。【结论】皮下注射M-CSF诱发的机械性痛觉超敏存在性别二态性,并与外周巨噬细胞扩增和伤害性通路敏化的差异性变化有关。【Objective】To investigate whether there exists gender differences in mechanical pain hypersensitivity in-duced by the subcutaneous injection of macrophage colony-stimulating factor(M-CSF)in normal mice and to explore the preliminary mechanism.【Methods】Thirty 10-week-old C 57 BL/6 J mice were randomly divided into three groups,(n=10mice/group,half male and half female).The albumin control group(BSA,0.3μg),low dose M-CSF group(L M-CSF,0.075μg)and high dose M-CSF group(H M-CSF,0.3μg)received 50μL BSA or M-CSF injected subcutaneously into the left medial thigh once daily for 3 consecutive days.Before and after drug administration,von-Frey mechanical sensitiv-ity test was used to detect the mechanical paw withdrawal threshold(PWT)in each group.Immunofluorescence was per-formed to examine the expression changes of Ionized calcium-binding adaptor molecule 1(Iba 1)in skin,calcitonin gene related peptide(CGRP)and phosphorylated ERK 1/2(p-ERK)in L 5-L 6 DRG and lumbar spinal dorsal horn.【Results】In female mice,only high dose of M-CSF caused mechanical allodynia,whereas in male mice both doses produced marked allodynia.Mechanically,high-dose M-CSF induced massive aggregation of subcutaneous macrophages(marked by Iba 1)in male and female mice,but more dramatic dependence in female mice.Similar gender differences were also found in the increase of p-ERK and CGRP expression in dorsal root ganglion(DRGs).Notably,CGRP expression was es-pecially elevated in the fibers of DRG in male mice.Correspondingly,the expressions of p-ERK and CGRP+terminals in the superficial spinal dorsal horn of male mice were significantly higher than those of female mice after M-CSF treatment.【Conclusion】Subcutaneous injection of M-CSF triggers sexual dimorphism in mechanical pain hypersensitivity,which is related with differential changes in peripheral macrophage expansion and sensitization of the nociceptive pathway.

关 键 词：痛觉超敏 性别二态性 巨噬细胞 巨噬细胞集落刺激因子 降钙素基因相关肽 

分 类 号：R441.1[医药卫生—诊断学]