目 录缩略语表 .............................................. 2 中文摘要 .............................................. 4 ABSTRACT .............................................. 6 前 言 .............................................. 8 正 文 ............................................. 14 第一部分 小鼠肺纤维化模型的制备 ..................... 14 实验材料 ...................................... 14 实验方法 ...................................... 17 结果 .......................................... 20 讨 论 ........................................ 25 结论 .......................................... 26 第二部分 细胞输注对百草枯诱导小鼠肺纤维化的影响 ...... 27 实验材料 ...................................... 27 实验方法 ...................................... 28 结果 .......................................... 30 讨 论 ........................................ 37 结论 .......................................... 40 参考文献 ............................................. 41 附录 ................................................. 47 致谢 ................................................. 48 综述 ................................................. 491 缩略语表缩略词PQ英文全称 中文全称Paraquat 百草枯NADPHAECⅠAECⅡ7AADcfustriphosphopyridine nucleotidetype I alveolar epithelial cellstypeⅡalveolar epithelial cells7-Amino-Actinomycin Dcolony forming unitsBleomycin烟酰胺腺嘌呤二核苷酸磷酸肺泡Ⅰ型上皮细胞肺泡Ⅱ型上皮细胞7氨基放线菌素D集落形成单位博来霉素BLMBone marrow-derived mesenchymalBMSCs 骨髓间充质干细胞stem cellsUC-MSCsEGFumbilical cord mesenchymal stem cellsepidermal growth factorfibroblast growth factor -10hepatic growth factor脐带间充质干细胞表皮生长因子FGF-10HGF成纤维细胞生长因子10肝细胞生长因子内皮细胞EC endotheliocyteTGFβ-1KGFtransforming growth factor β-1keratinocyte growth factorbovine serum albumin转化生长因子β-1角质细胞生长因子牛血清白蛋白BSAPBS phosphate buffered saline 磷酸盐缓冲液2 EMTId2epithelial-Mesenchymal transitioninhibitor of differentiation 2polyunsa turated fatty acidsuperoxide dismutase上皮间质细胞转化分化抑制因子2多不饱和脂肪酸超氧化物歧化酶谷胱甘肽PUFASODGSH glutathioneES embryonic stem cell 胚胎干细胞LD50 median lethal dose 半数致死剂量3 细胞输注治疗百草枯中毒性肺纤维化的可行性研究摘要 目的：通过腹腔注射百草枯构建小鼠肺纤维化模型；探讨肺组织细胞亚群输注对百草枯诱导肺纤维化的影响及可能机制。
方法：一、腹腔注射不同浓度百草枯制备小鼠肺纤维化病理模型，筛选出最佳造模剂量 1、 动物分组：正常雌性 C57BL/6小鼠 60只，随机分为 6组，10只/组，分别为正常对照组，百草枯给药 30mg/kg组，40mg/kg组，50mg/kg组，60mg/kg组和 80mg/kg组；2、模型制备：各染毒组予一次性腹腔注射不同浓度百草枯溶液，正常对照组予腹腔注射等容积无菌生理盐水；3、观察指标：染毒后 28内各组小鼠精神状态、存活情况、体重及肺组织病理改变，并计算其半数致死剂量（LD50值）及各组肺纤维化Ashcroft评分二、探讨肺组织细胞亚群输注对百草枯诱导小鼠肺纤维化的影响及可能机制 1、动物分组: 正常雌性 C57BL/6小鼠 90只，随机分为 6 组，分别为正常对照组、百草枯 50mg/kg 模型组、全肺细胞输注组、上皮细胞输注组、间质细胞输注组；2、模型制备及肺组织细胞亚群流式分选：模型组及细胞治疗组予一次性腹腔注射百草枯溶液（50mg/kg），给药后 24h 各细胞输注组予尾静脉输注正常小鼠各肺组织细胞亚群（流式细胞仪分选），正常对照组予腹腔注射等容积生理盐水，24h 后予尾静脉输注等容积无菌生理盐水；3、观察指标：细胞输注后 28内各组小鼠精神状态、存活情况及肺组织病理变化（HE、Masson染色），并根据HE染色结果进行肺纤维化 Ashcroft评级。
结果：一、百草枯中毒致肺纤维化造模剂量筛选：至观察期28天，小鼠一次性腹腔注射百草枯溶液的LD50为55.19mg/kg；各染毒组均出现不同程度的肺纤维化改变，差异有统计学意义（P0.05）结论：40mg/kg和50mg/kg均为较合适的造模剂量，研究者可根据不同的研究目的选择相应的染毒剂量；肺组织全肺细胞、上皮细胞、间质细胞输注可减轻或延缓百草枯诱导的小鼠肺纤维化，尤其是特定的单一肺组织细胞亚群输注亦能发挥其修复效应关键词：肺纤维化 百草枯 肺组织细胞亚群5 Feasibility Study of Cell infusion on Treatment of PulmonaryFibrosis Induced by ParaquatAbstractObjective: By establish a model of pulmonary fibrosis induced by intraperitonealinjection of paraquat in mice, to investigate the effects of different lung tissue cellpopulations on paraquat-induced pulmonary fibrosis and to study the possiblemechanism of its effects.Methods:Ⅰ.to select the optimal paraquat dosage by a single intraperitoneal injectionof different concentrations of paraquat to establish a model of pulmonary fibrosis.1.Animal grouping: 60 healthy female C57BL/6 mice were randomly divided into sixgroups according to the paraquat dosage:the saline control group,30mg/kg paraquatgroup, 40mg/kg paraquat group,50mg/kg paraquat group,60mg/kg paraquat group and80mg/kg paraquat group. Each group had 10 mice;2. Model Preparation: the mice inmodel groups were injected intraperitoneal 30mg/kg、40mg/kg、50mg/kg、60mg/kg and80mg/kg paraquat while the control group were administrated 0.9% sodium chloridesolution;3.Observing indexes：mental status、life-span、body weight changes and thepathologic changes were observed,and the LD50 was calculated , the sections afterHE staining were evaluated for the formation of pulmonary fibrosis by using Ashcroftscoring system.Ⅱ.to investigate the effects of different lung tissue cell populations onparaquat-induced pulmonary fibrosis.1. Animal grouping: 90 health female C57BL/6mice, were randomly divided into 6 groups,respectively,the normal control group,the 50mg/kg paraquat group, the 7AAD- infusion group,CD45-/CD31-/Sca-1-/EpCAM+infusion group,CD45-/CD31-/EpCAM-/Sca-1+ cell infusion group;2.Model Preparationand flow sorting of lung tissue cell subsets: Each group were administrated a singleintraperitoneal injection of solution(50mg/kg),After 24h, the mice in infusion groupwere administrated different lung tissue cell subsets respectively (FACS sorting) whilethe control group were administrated 0.9% sodium ch。