髓系细胞特异性敲除 Capns1 可防止巨噬细胞向 M1 表型极化，并减少博来霉素诱导的系统性硬皮病模型中的间质性肺病。
Myeloid cell-specific deletion of Capns1 prevents macrophage polarization toward the M1 phenotype and reduces interstitial lung disease in the bleomycin model of systemic sclerosis.
机构信息
Department of Dermatology, Zhongshan Hospital, Fudan University, Xuhui District, 180 Fenglin Road, Shanghai, 200032, People's Republic of China.
Department of Dermatology, Huashan Hospital, Fudan University, 12 Wulumuqi Middle Road, Shanghai, 200040, People's Republic of China.
出版信息
Arthritis Res Ther. 2022 Jun 21;24(1):148. doi: 10.1186/s13075-022-02833-7.
BACKGROUND
Calpains are a family of calcium-dependent thiol proteases that participate in a wide variety of biological activities. In our recent study, calpain is increased in the sera of scleroderma or systemic sclerosis (SSc). However, the role of calpain in interstitial lung disease (ILD) has not been reported. ILD is a severe complication of SSc, which is the leading cause of death in SSc. The pathogenesis of SSc-related ILD remains incompletely understood. This study investigated the role of myeloid cell calpain in SSc-related ILD.
METHODS
A novel line of mice with myeloid cell-specific deletion of Capns1 (Capns1-ko) was created. SSc-related ILD was induced in Capns1-ko mice and their wild-type littermates by injection 0.l mL of bleomycin (0.4 mg/mL) for 4 weeks. In a separate experiment, a pharmacological inhibitor of calpain PD150606 (Biomol, USA, 3 mg/kg/day, i.p.) daily for 30 days was given to mice after bleomycin injection on daily basis. At the end of the experiment, the animals were killed, skin and lung tissues were collected for the following analysis. Inflammation, fibrosis and calpain activity and cytokines were assessed by histological examinations and ELISA, and immunohistochemical analyses, western blot analysis and Flow cytometry analysis.
RESULTS
Calpain activities increased in SSc-mouse lungs. Both deletion of Capns1 and administration of PD150606 attenuated dermal sclerosis as evidenced by a reduction of skin thickness and reduced interstitial fibrosis and inflammation in bleomycin model of SSc mice. These effects of reduced calpain expression or activity were associated with prevention of macrophage polarization toward M1 phenotype and consequent reduced production of pro-inflammatory cytokines including TNF-α, IL-12 and IL-23 in lung tissues of Capns1-ko mice with bleomycin model of SSc. Furthermore, inhibition of calpain correlated with an increase in the protein levels of PI3K and phosphorylated AKT1 in lung tissues of the bleomycin model of SSc mice.
CONCLUSIONS
This study for the first time demonstrates that the role of myeloid cell calpain may be promotion of macrophage M1 polarization and pro-inflammatory responses related PI3K/AKT1 signaling. Thus, myeloid cell calpain may be a potential therapeutic target for bleomycin model of SSc-related ILD.
背景
钙蛋白酶是一类依赖钙的硫醇蛋白酶家族，参与多种生物活性。在我们最近的研究中，硬皮病或系统性硬皮病（SSc）患者血清中的钙蛋白酶增加。然而，钙蛋白酶在间质性肺病（ILD）中的作用尚未报道。ILD 是 SSc 的严重并发症，是 SSc 的主要死亡原因。SSc 相关 ILD 的发病机制尚不完全清楚。本研究探讨了髓细胞钙蛋白酶在 SSc 相关 ILD 中的作用。
方法
创建了髓细胞特异性钙蛋白酶 1（Capns1）缺失的新型小鼠（Capns1-ko）。通过注射 0.1 mL 博来霉素（0.4mg/mL），4 周后在 Capns1-ko 小鼠及其野生型同窝仔鼠中诱导 SSc 相关 ILD。在另一个实验中，在博来霉素注射后每天腹腔内给予钙蛋白酶的药理学抑制剂 PD150606（Biomol，美国，3mg/kg/天）30 天。实验结束时，处死动物，采集皮肤和肺组织进行以下分析。通过组织学检查和 ELISA、免疫组织化学分析、western blot 分析和流式细胞术分析评估炎症、纤维化和钙蛋白酶活性以及细胞因子。
结果
钙蛋白酶活性在 SSc 小鼠肺中增加。Capns1 缺失和 PD150606 给药均减轻了博来霉素诱导的 SSc 小鼠的皮肤硬化，表现为皮肤厚度减少，间质纤维化和炎症减少。钙蛋白酶表达或活性降低的这些作用与防止巨噬细胞向 M1 表型极化以及随后减少肺组织中 TNF-α、IL-12 和 IL-23 等促炎细胞因子的产生有关。此外，博来霉素诱导的 SSc 小鼠肺组织中钙蛋白酶的抑制与 PI3K 和磷酸化 AKT1 蛋白水平的增加相关。
结论
本研究首次证明，髓细胞钙蛋白酶的作用可能是促进巨噬细胞 M1 极化和与 PI3K/AKT1 信号相关的促炎反应。因此，髓细胞钙蛋白酶可能是博来霉素诱导的 SSc 相关 ILD 的潜在治疗靶点。
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