Ren Ying, Yang Hui, Zhu Ping, Fan Chun-mei, Wang Yan-hong, Li Jia, Liu Hui
Department of Clinical Immunology, Xijing Hospital, Fourth Military Medical University, Xi'an, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 Mar;28(3):232-6.
To observe the therapeutic effect of cyclosporine A (CsA) on bleomycin (BLM) induced pulmonary fibrosis and to investigate its mechanism.
One hundred and twenty C57BL/6 female mice were divided randomly into five groups: BLM model group, control saline group, CsA30 mg treatment group, CsA50 mg treatment group and control treatment group. Treatment groups and model groups were administrated BLM intratracheally to induce interstitial pulmonary disease model, with control saline group administrated with equal volume of normal saline instead. Mice in treatment groups were intraperitoneal injected with CsA, while control treatment group were injected with equal volume of normal saline instead. On the 4th, 7th and 14th day after administration, 8 mice of each group were sacrificed, and the peripheral blood was obtained to count total leucocytes with counting chamber and quantify CD4(+); T cells, CD14(+); monocytes and CD19(+); B cells by flow cytometry (FCM). Bronchoalveolar levage fluid was harvested for cell counting and Giemsa staining. Lung tissues were harvested for immunohistochemical staining and pathological examination.
The quantity of total leucocyte was higher in BLM model group than those in control saline group.The proportion of CD14(+); T cells and CD19(+);B cells in BLM model group were increased markedly than those in control saline group on the 4th, 7th and 14th day post BLM. With CsA treatment, The proportion of CD14(+); T cells was lower than BLM model group at the same time point, especially on the 4th day. The proportion of CD19(+); B cells were significantly lower than those of BLM model group at the same time point(7 d, 14 d). The total and classification of cells of BLM model group were increased markedly than those in control saline group, and decreased obviously in the treatment groups at the same time point. Examination of lung tissues: With the prolonged time of BLM administration, it showed wider alveolar septum, more collagen deposition, as well as more infiltrating inflammatory cells which consisted of generous lymphocyte and few mononuclear macrophages than those in saline control group. With the prolonged time of CsA injection, the interstitial pulmonary inflammation was remissive, and there was less fibroblast infiltration and collagen deposition in pulmonary interstitium and periphery of bronchiole. Alveolar epithelial cells, bronchiolar epithelial cells, mononuclear macrophages, neutrophils and lymphocytes were demonstrated to express CD147, there was higher CD147 expression in BLM model group than those in CsA treatment groups.
CsA may heal BLM induced interstitial pulmonary disease by blocking CD147-CypA interaction, then decreasing chemotaxis for the immunocyte, and reducing migration of immunocytes to the lung and collagen deposition in the lung.
观察环孢素A（CsA）对博来霉素（BLM）诱导的肺纤维化的治疗效果并探讨其机制。
将120只C57BL/6雌性小鼠随机分为五组：BLM模型组、生理盐水对照组、CsA 30mg治疗组、CsA 50mg治疗组和对照治疗组。治疗组和模型组经气管内给予BLM以诱导间质性肺病模型，生理盐水对照组给予等体积的生理盐水。治疗组小鼠腹腔注射CsA，对照治疗组注射等体积的生理盐水。给药后第4、7和14天，每组处死8只小鼠，采集外周血用血细胞计数板计数白细胞总数并通过流式细胞术（FCM）定量CD4(+)；T细胞、CD14(+)；单核细胞和CD19(+)；B细胞。收集支气管肺泡灌洗液进行细胞计数和吉姆萨染色。采集肺组织进行免疫组织化学染色和病理检查。
BLM模型组白细胞总数高于生理盐水对照组。BLM后第4、7和14天，BLM模型组CD14(+)；T细胞和CD19(+)；B细胞的比例明显高于生理盐水对照组。经CsA治疗后，同一时间点CD14(+)；T细胞的比例低于BLM模型组，尤其是在第4天。同一时间点（7天、14天）CD19(+)；B细胞的比例明显低于BLM模型组。BLM模型组细胞总数和分类明显高于生理盐水对照组，同时治疗组明显降低。肺组织检查：随着BLM给药时间延长，肺泡间隔增宽，胶原沉积增多，浸润的炎性细胞比生理盐水对照组更多，由大量淋巴细胞和少量单核巨噬细胞组成。随着CsA注射时间延长，间质性肺炎缓解，肺间质和细支气管周围成纤维细胞浸润和胶原沉积减少。肺泡上皮细胞、细支气管上皮细胞、单核巨噬细胞、中性粒细胞和淋巴细胞均表达CD147，BLM模型组CD147表达高于CsA治疗组。
CsA可能通过阻断CD147-CypA相互作用，进而降低免疫细胞趋化性，并减少免疫细胞向肺内迁移及肺内胶原沉积，从而治愈BLM诱导的间质性肺病。
