放射性肺纤维化小鼠模型建立及治疗效果评价

Mouse model establishment and therapeutic evaluation of radiation-induced pulmonary fibrosis

摘要

目的 基于基因编辑技术构建人CD147(hCD147)小鼠放射性肺纤维化模型, 探究靶向CD147的人源化抗体———美珀珠单抗抑制肺纤维化进展的作用, 为临床放射性肺损伤治疗提供实验依据。方法 通过⁶⁰Co辐照建立小鼠放射性肺纤维化模型, 收集辐照后第10日及未辐照组(第0日)小鼠肺组织样本进行转录组测序, 明确辐照介导的肺部炎性损伤特征; 构建hCD147小鼠放射性肺纤维化模型, 将小鼠分为两组, 于辐照后第2、4和6日分别腹腔注射3 mg/kg的IgG(IgG组)和美珀珠单抗(美珀珠单抗组), 通过体质量监测、组织病理染色及Western blotting评估抗体治疗效果。结果 成功建立⁶⁰Co辐照的小鼠放射性肺纤维化模型, 辐照后第10日小鼠体质量降幅明显, 肺组织呈急性炎性损伤, 转录组测序显示未辐照和辐照后第10日小鼠肺组织的差异表达基因与免疫炎性反应密切相关, 与未辐照(第0日)相比, 辐照后第10日肺组织TNF-α、IL-1β水平显著升高; Masson染色实验显示, 辐照后第30日至第120日肺组织呈现慢性纤维化进展特征, 胶原纤维沉积增多; 与未辐照(第0日)相比, 辐照后第120日小鼠肺组织α-SMA表达升高。通过基因编辑技术成功构建hCD147小鼠并进行⁶⁰Co照射, 与IgG组相比, 美珀珠单抗组小鼠体质量恢复更佳, 肺组织炎性损伤及胶原纤维沉积较少, α-SMA表达水平较低。结论 本研究成功建立⁶⁰Co辐照诱导的放射性肺纤维化小鼠模型, 小鼠肺组织表现为急性炎性损伤向慢性纤维化演进的病理进程, 以CD147为靶点的美珀珠单抗可减轻炎性肺损伤、抑制纤维化进程, 为放射性肺纤维化治疗提供了潜在靶点。

Abstract

Objective To investigate the inhibitory effect of meplazumab, a humanized antibody targeting CD147, on the progression of pulmonary fibrosis by constructing a human CD147 (hCD147) mouse model of radiation-induced pulmonary fibrosis based on gene editing technology, thereby providing experimental evidence for the clinical treatment of radiation-induced lung injury. Methods A mouse model of radiation-induced pulmonary fibrosis was established by ⁶⁰Co irradiation. Lung tissue samples from mice on the 10th day after irradiation and the non-irradiated group (day 0) were collected for transcriptome sequencing to clarify the characteristics of radiation-mediated pulmonary inflammatory injury. The hCD147 mouse model of radiation-induced pulmonary fibrosis was constructed and divided into two groups, and on the 2nd, 4th and 6th days after irradiation, 3 mg/kg of IgG (IgG group) and meplazumab (meplazumab group) were intraperitoneally injected, respectively. The therapeutic effect of the antibody was evaluated by body mass monitoring, histopathological staining, and Western blotting experiments. Results A mouse model of radiation-induced pulmonary fibrosis induced by ⁶⁰Co irradiation was successfully established. On the 10th day post-irradiation, the mice exhibited a significant body mass decline and acute inflammatory injury in their lung tissues. Transcriptome sequencing revealed that the differentially expressed genes in lung tissues between unirradiated and irradiated mice on the 10th day after irradiation were closely associated with immune-inflammatory responses. Compared with day 0, the levels of TNF-α and IL-1β in the lung tissues of irradiated mice were significantly elevated on the 10th day. Masson staining results demonstrated that from the 30th to the 120th day post-irradiation, the lung tissues displayed progressive chronic fibrosis characteristics, accompanied by increased collagen fiber deposition. Additionally, the expression level of α-SMA in the lung tissues of irradiated mice on the 120th day was significantly higher than that on day 0. Furthermore, hCD147 mice were successfully constructed via gene editing technology and then subjected to ⁶⁰Co irradiation. Compared with the IgG group, mice in the meplazumab group showed better body mass recovery, reduced pulmonary inflammatory injury and collagen fiber deposition, as well as lower α-SMA expression level. Conclusion This study successfully establishes a mouse model of radiation-induced pulmonary fibrosis via ⁶⁰Co irradiation. The lung tissues of the mice exhibit a pathological progression from acute inflammatory injury to chronic fibrosis. Meplazumab targeting CD147 alleviates pulmonary inflammatory injury and inhibits the fibrotic process, thereby providing a potential therapeutic target for radiation-induced pulmonary fibrosis.

关键词

人CD147小鼠 / 放射性肺纤维化 / 炎性损伤 / 美珀珠单抗 / 免疫应答 / 胶原纤维 / 疗效评价 / 治疗靶点

Key words

human CD147 mouse / radiation-induced pulmonary fibrosis / inflammatory injury / meplazumab / immune response / collagen fiber / efficacy evaluation / therapeutic target

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基金资助

XX实验动物专项(SYDW_KY[2021]15)

中国科学技术协会青年人才托举工程项目(YESS20200011)