Huang Yali, Zeng Wen, Liao Xiaoling, Qin Fang, Ma Xuejia, Pan Jie, Li Guangting, Tang Kaijiang, Lei Ling
Department of Rheumatology and Immunology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Province; and Department of Rheumatology and Immunology, The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, Guangxi Province, China.
Department of Rheumatology and Immunology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Province, China.
Clin Exp Rheumatol. 2025 Aug;43(8):1398-1407. doi: 10.55563/clinexprheumatol/1t3s0i. Epub 2025 Mar 19.
We investigated the impact of IL-35 and IL-10 on the immune response and pulmonary fibrosis using a bleomycin (BLM)-induced SSc mouse model.
BLM was administered subcutaneously to Balb/c mice and either mouse recombinant (rm)IL-35, rmIL-10 or neutralising antibody of IL-35 and IL-10 was injected intraperitoneally after BLM administration. Lung fibrosis was assessed by the pathological alterations, hydroxyproline content, and the collagen I and α-SMA mRNA expression. The expression of immune cells and their related factors were respectively measured by flow cytometry and ELISA. Western blot was used to measure STAT3 pathway expression.
Compared with controls, BLM exposure induced increased Ashcroft ratings, hydroxyproline and lung collagen I and α-SMA expression, which was lessened by rmIL-35 or rmIL-10 intervention, while it did not change after blocking IL-35 and IL-10. BLM exposure increased IL-4 and IL-17A expression in bronchoalveolar lavage (BAL) supernatant, which was downregulated by rmIL-35 or rmIL-10 administration. Compared with the BLM group, the RmIL-35 and rmIL-10 group both downregulated Th2/nTreg and Th17/nTreg percentage, while increased Treg cell proportion in the spleen. Moreover, the spleen iTr35 cell ratio was negatively correlated with BAL supernatant IL-17A and IL-4 levels and lung collagen I and α-SMA expression. Further pathway analysis revealed that rmIL-35 administration decreased the phosphorylation of STAT3 compared with the BLM group.
Our findings suggest that IL-35 and IL-10 might alleviate pulmonary inflammation and fibrosis via upregulating the proportion of Treg cells and reducing BAL supernatant IL-17A and IL-4 levels in a bleomycin-induced SSc mouse model.
我们使用博来霉素（BLM）诱导的系统性硬化症（SSc）小鼠模型，研究白细胞介素-35（IL-35）和白细胞介素-10（IL-10）对免疫反应和肺纤维化的影响。
将BLM皮下注射给Balb/c小鼠，并在注射BLM后腹腔注射小鼠重组（rm）IL-35、rmIL-10或IL-35和IL-10的中和抗体。通过病理改变、羟脯氨酸含量以及I型胶原蛋白和α-平滑肌肌动蛋白（α-SMA）mRNA表达评估肺纤维化。分别通过流式细胞术和酶联免疫吸附测定（ELISA）测量免疫细胞及其相关因子的表达。使用蛋白质免疫印迹法测量信号转导和转录激活因子3（STAT3）通路的表达。
与对照组相比，BLM暴露导致阿什克罗夫特评分、羟脯氨酸以及肺I型胶原蛋白和α-SMA表达增加，rmIL-35或rmIL-10干预可减轻这些变化，而阻断IL-35和IL-10后则无变化。BLM暴露增加了支气管肺泡灌洗（BAL）上清液中IL-4和IL-17A的表达，rmIL-35或rmIL-10给药可使其下调。与BLM组相比，RmIL-35和rmIL-10组均下调了脾脏中Th2/天然调节性T细胞（nTreg）和Th17/nTreg的百分比，同时增加了调节性T细胞（Treg）的比例。此外，脾脏诱导性Tr35细胞比例与BAL上清液中IL-17A和IL-4水平以及肺I型胶原蛋白和α-SMA表达呈负相关。进一步的通路分析显示，与BLM组相比，rmIL-35给药降低了STAT3的磷酸化水平。
我们的研究结果表明，在博来霉素诱导的SSc小鼠模型中，IL-35和IL-10可能通过上调Treg细胞比例和降低BAL上清液中IL-17A和IL-4水平来减轻肺部炎症和纤维化。
