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目的 采用趋化因子受体即CXCR3基因敲除小鼠 ,探讨CXCR3在博莱霉素诱导的肺损伤及纤维化中的作用。方法 采用基因打靶技术得到无CXCR3基因小鼠 6 2只 ,同时选择性别、年龄和体重配对的野生型小鼠 4 8只 ,随机分入不同的实验组及对照组。大鼠气管内注入博莱霉素0 0 2 5U。实验动物按要求处死后 ,取无血的肺组织切片 ,经 10 %甲醛固定后 ,用苏木精 伊红 (HE)和Massontrichrome染色 ,分别观察实验动物肺炎症和纤维化的程度 ;用浓盐酸消化肺组织提取羟脯氨酸并定量来表示肺胶原的含量 ;用磷酸盐缓冲液 (PBS)灌洗肺组织 ,酶联免疫吸附 (ELISA)法测定灌洗液中肿瘤坏死因子α(TNF α)、白细胞介素 5 (IL 5 )及转化生长因子 β(TGF β)的浓度 ;用流式细胞仪检测T细胞亚群。结果 气管内注入博莱霉素后 14d ,CXCR3基因敲除小鼠肺组织炎症程度及纤维化程度较野生型小鼠明显减轻。肺组织炎症程度评分和羟脯氨酸含量 (左肺 )在CXCR3基因敲除小鼠分别为 3 92± 0 37和 (6 7 0± 2 4 2 ) μg ,在野生型小鼠则分别为 5 33± 0 34和 (2 11 5± 2 4 2 )μg ,两组比较差异有统计学意义 (P分别 <0 0 5 ,<0 0 1) ;TGF β水平在CXCR3基因敲除小鼠为(2 2 11± 2 89) μg/L ,在野生型小鼠为 (5 388± 10 71) μg/L ,
Objective To investigate the role of CXCR3 in bleomycin-induced lung injury and fibrosis in CXCR3-knockout mice, a chemokine receptor. Methods Forty-two mice without CXCR3 gene were obtained by gene targeting technique. Forty-eight wild-type mice of sex, age and body weight were randomly assigned to different experimental and control groups. Rat bleomycin intratracheal injection 0 0 2 5U. After the animals were killed according to the requirements, blood-free lung tissue sections were taken and stained with hematoxylin and eosin (HE) and Massontrichrome after being fixed with 10% formaldehyde. The degree of lung inflammation and fibrosis in the experimental animals were observed respectively. Lung tissues were digested with hydroxyproline and quantitated to express lung collagen. Lung tissue was perfusion with phosphate buffered saline (PBS). Tumor necrosis factor-α (TNFα) in the lavage fluid was measured by enzyme linked immunosorbent assay (ELISA) ), Interleukin 5 (IL 5) and transforming growth factor β (TGF β) were detected by flow cytometry. T cell subsets were detected by flow cytometry. Results 14 days after intratracheal instillation of bleomycin, lung inflammation and fibrosis in CXCR3 knockout mice were significantly lower than those in wild type mice. Lung tissue inflammation score and hydroxyproline content (left lung) were CX3D3 knockout mice were 3 92 ± 0 37 and (67 0244) μg, respectively, in wild-type mice were 5 33 ± 0 34 and (2 11 5 ± 2 4 2) μg, respectively, with significant difference between the two groups (P <0 05 and <0 0 1, respectively); TGF β levels in CXCR3 knockout mice were ( 2 2 11 ± 2 89) μg / L, (5 388 ± 10 71) μg / L in wild-type mice,