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ERβ介导的雌二醇信号促进小鼠急性骨骼肌炎缓解与肌纤维修复

谢俊毅¹, 蔡祺晖¹, 牛晓璐¹, 王楠¹, 胡欣雨¹, 黄晓蕾¹, 王瑞雪², 陈哲³, 廖钊宏^1,4,*

¹佛山大学医学部医学技术系，佛山 528225；²佛山大学医学部基础医学系，佛山 528225；³佛山大学医学部护理学系，佛山528225；⁴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人体解剖学教研室，广州 510515

摘要

创伤应激可导致性激素水平异常，其中雌激素水平升高与患者预后改善相关，但其对急性骨骼肌炎炎症反应及修复进程的影响机制尚不明确。本文旨在探究雌二醇(estradiol, E2)信号对心肌毒素(cardiotoxin, CTX)诱导的小鼠急性骨骼肌炎缓解与肌纤维修复的影响。采用C57BL/6 小鼠建立急性骨骼肌炎模型，通过雌鼠卵巢去势(ovariectomized, OVX)降低内源性E2水平，并用β-雌二醇(β-Estradiol)或雌激素受体β (estrogen receptor β, ERβ)特异性拮抗剂4-[2-苯基-5,7-双(三氟甲基)吡唑并[1,5-a]嘧啶-3-基]苯酚(4- [2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a] -pyrimidin-3-yl]phenol, PHTPP)进行干预；分化培养C2C12 细胞为成熟肌管后，用干扰素-γ (interferon γ, IFN-γ)联合β-Estradiol 或PHTPP处理。用ELISA检测各组小鼠血清E2水平，用HE染色检测损伤肌炎性细胞浸润程度，用免疫荧光染色检测损伤肌内ERβ、胚胎肌球蛋白重链(embryonic myosinheavy chain, eMHC)、CD45 和分化肌管内生肌决定因子(myogenic determining factor, MyoD)、肌细胞生成素(myogenin)表达水平，用流式细胞术检测损伤肌内CD45⁺炎性细胞比例，用Western blot 检测分化肌管内小鼠主要组织相容性复合体Ⅰ类分子(major histocompatibility complex class I molecules, MHC-I) H-2Kb、小鼠主要组织相容性复合体II 类分子(majorhistocompatibility complex class II molecules, MHC-II) H2-Eα、Toll 样受体3 (Toll-like receptor 3, TLR3)，以及ERβ 蛋白表达水平，用RT-qPCR 法检测损伤肌及分化肌管内ERα、ERβ、G蛋白耦联受体30 (G protein-coupled receptor 30, GPR30)、白细胞介素1β (interleukin-1β, IL-1β)、IL-6、IL-10，及单核细胞趋化蛋白1 (monocyte chemoattractant protein-1, MCP-1) mRNA表达水平。结果显示，在急性骨骼肌炎条件下，雌鼠损伤肌炎症轻于雄鼠，且血清E2水平显著高于雄鼠；损伤肌及炎性肌管中ERβ 表达显著上调，而ERα 和GPR30 无明显变化。OVX导致雌鼠血清E2水平降低、损伤肌炎症加重及CD45⁺炎性细胞浸润增加，β-Estradiol 处理可逆转OVX雌鼠的上述改变，而PHTPP的作用与OVX相同，除了不能影响血清E2水平。在炎性肌管上，β-Estradiol 上调ERβ 表达水平，下调免疫分子H-2Kᵇ、H2-Eα、TLR3，以及促炎因子IL-1β、IL-6、MCP-1 表达，上调抗炎因子IL-10 表达，并上调MyoD和myogenin 表达，而PHTPP 的作用和β-Estradiol 相反。OVX小鼠损伤肌中eMHC表达下调，β-Estradiol 可恢复其表达。以上结果提示，在急性骨骼肌炎中，E2可能通过与肌纤维ERβ 结合抑制损伤肌内炎性细胞聚集，调控炎性肌纤维内在免疫行为，从而促进炎症缓解与肌纤维再生修复，为急性骨骼肌炎的临床治疗提供了新思路。

关键词： 雌二醇信号; 雌二醇受体β; 急性骨骼肌炎; 免疫行为; 肌纤维修复

ERβ-mediated estrogen signaling promotes the alleviation of inflammation and myofiber repair in cardiotoxin-induced acute skeletal myositis in mice

XIE Jun-Yi¹, CAI Qi-Hui¹, NIU Xiao-Lu¹, WANG Nan¹, HU Xin-Yu¹, HUANG Xiao-Lei¹, WANG Rui-Xue², CHEN Zhe³, LIAO Zhao-Hong^1,4,*

¹Department of Laboratory Medicine, School of Medicine, Foshan University, Foshan 528225, China；²Department of Basic Medicine, School of Medicine, Foshan University, Foshan 528225, China；³Department of Nursing Science, School of Medicine, Foshan University, Foshan 528225, China；⁴Guangdong Provincial Key Laboratory of Tissue Construction and Testing, Department of Anatomy, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China

Abstract

Traumatic stress often leads to abnormalities in sex hormone levels, among which elevated estrogen levels are associated with improved patient prognosis. However, the mechanism by which estrogen affects the inflammatory response and repair process in acute skeletal myositis remains unclear. This study aimed to investigate the effects of estradiol (E2) signaling on the resolution of acute skeletal myositis and muscle fiber repair in mice induced by cardiotoxin (CTX). An acute skeletal myositis model was established in C57BL/6 mice. Endogenous E2 levels were reduced by ovariectomy (OVX) in female mice, and interventions were performed with β-Estradiol or the estrogen receptor β (ERβ)-specific antagonist 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP). Differentiated C2C12 myotubes were treated with interferon γ (IFN-γ) in combination with β-Estradiol or PHTPP. Serum E2 levels were measured by ELISA. Inflammatory cell infiltration in injured muscle was assessed by HE staining. Immunofluorescence staining was used to detect the expression levels of ERβ, embryonic myosin heavy chain (eMHC), CD45, myogenic determining factor (MyoD) and myogenin. Proportion of CD45+ inflammatory cells in injured muscle was detected by flow cytometry. Western blot was used to detect protein expression levels of mouse major histocompatibility complex class I molecules (MHC-I) H-2Kb, major histocompatibility complex class II molecules (MHC-II) H2-Eα, Toll-like receptor 3 (TLR3), and ERβ in differentiated myotubes. RT-qPCR was used to measure mRNA expression levels of ERα, ERβ, G protein-coupled receptor 30 (GPR30), interleukin-1β (IL-1β), IL-6, IL-10, and monocyte chemoattractant protein-1 (MCP-1) in injured muscle and differentiated myotubes. The results showed that, in acute skeletal myositis, injured muscle of female mice exhibited milder inflammation compared to male mice, and serum E2 levels were significantly higher in the female mice. Meanwhile, expression of ERβ was significantly up-regulated in injured muscle and inflammatory myotubes, whereas no significant changes were observed in ERα and GPR30. OVX decreased serum E2 level, exacerbated inflammation, and increased infiltration of CD45+ inflammatory cells in injured muscle, whereas these effects were reversed by β-Estradiol treatment. The effects of PHTPP were the same as those of OVX, except that it didn't affect serum E2 levels. In inflammatory myotubes, β-Estradiol up-regulated the protein expression level of ERβ, down-regulated the expression levels of immune molecules H-2Kb, H2-Eα, TLR3, and pro-inflammatory factors IL-1β, IL-6, MCP-1, up-regulated the expression of anti-inflammatory factor IL-10, and up-regulated the expression of MyoD and myogenin. The effects of PHTPP were opposite to those of β-Estradiol. The expression of eMHC in the injured muscle of OVX mice was down-regulated, while β-Estradiol restored its expression. These results suggest that in acute skeletal myositis, E2 inhibits the accumulation of inflammatory cells in the injured muscle by binding to ERβ in muscle fibers, regulates the intrinsic immune behavior of inflammatory myofibers, thereby promoting inflammation resolution and muscle fiber regeneration and repair, providing new insights for the clinical treatment of acute skeletal myositis.

Key words: estrogen signaling; estrogen receptor β; acute skeletal myositis; immune behavior; myofiber repair

收稿日期：　　录用日期：

通讯作者：廖钊宏　　E-mail:

DOI: 10.13294/j.aps.2026.0049

引用本文：

谢俊毅, 蔡祺晖, 牛晓璐, 王楠, 胡欣雨, 黄晓蕾, 王瑞雪, 陈哲, 廖钊宏. ERβ介导的雌二醇信号促进小鼠急性骨骼肌炎缓解与肌纤维修复[J]. 生理学报 2026; 78 (3): 631-641.

XIE Jun-Yi, CAI Qi-Hui, NIU Xiao-Lu, WANG Nan, HU Xin-Yu, HUANG Xiao-Lei, WANG Rui-Xue, CHEN Zhe, LIAO Zhao-Hong. ERβ-mediated estrogen signaling promotes the alleviation of inflammation and myofiber repair in cardiotoxin-induced acute skeletal myositis in mice. Acta Physiol Sin 2026; 78 (3): 631-641 (in Chinese with English abstract).