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大鼠染矽尘后不同时间重复移植人脐带间充质干细胞对肺纤维化的影响
引用本文:陈志军,沙焱,黄惠霞,谢英,张健杰,李智民. 大鼠染矽尘后不同时间重复移植人脐带间充质干细胞对肺纤维化的影响[J]. 职业卫生与应急救援, 2019, 37(4): 302. DOI: 10.16369/j.oher.issn.1007-1326.2019.04.002
作者姓名:陈志军  沙焱  黄惠霞  谢英  张健杰  李智民
作者单位:深圳市职业病防治院, 广东 深圳 518020
基金项目:深圳市科技研发资金技术攻关项目JSGG20160226161357949;深圳市知识创新计划项目JCYJ20140414110951762
摘    要: 目的 探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUCMSCs)在不同时间窗的重复移植对染矽尘大鼠肺纤维化的疗效。
方法 将48只健康雄性SD大鼠随机分为4组:对照组、矽肺模型组、预防性干预组和治疗性干预组(每组12只)。采用非暴露式气管插管内灌注法,对照组注入0.5 mL生理盐水,其余各组均注入0.5 mL二氧化硅(SiO2)混悬液(50 mg/mL),1周后重复1次。首次染尘后,预防性干预组在第1、7、14、21天,治疗性干预组在第28、35、42、49天,经尾静脉缓慢注射0.5 mL HUCMSCs细胞悬液(3.0×109/L)。在第70天统一处理动物,以HE染色和Masson染色,观察肺组织病理改变并进行Aschcroft肺纤维化评分,检测肺系数、肺组织羟脯氨酸(hydroxyproline,Hyp)及转化生长因子-β1(TGF-β1)、血清白细胞介素-6(IL-6)的水平。
结果 肺组织病理显示,对照组肺泡结构正常,其余各组肺脏有不同程度纤维化病变,矽肺模型组见细胞纤维性肉芽肿形成,预防性干预组和治疗性干预组与矽肺模型组比较,胶原纤维明显减少;Aschcroft评分、肺系数、肺组织Hyp和TGF-β1、血清IL-6比较,染尘各组均明显高于对照组(P < 0.01),而预防性干预组和治疗性干预组明显低于矽肺模型组(P < 0.01),预防性干预组和治疗性干预组之间以上指标差异均无统计学意义(P>0.05)。
结论 采用重复注射HUCMSCs的方法,无论在以肺泡炎为主的阶段还是在以纤维化为主的阶段进行干预,均可以有效减轻矽尘导致的大鼠肺纤维化。


关 键 词:脐带间充质干细胞   矽肺   肺纤维化   羟脯氨酸   转化生长因子-β1   血清白细胞介素-6
收稿时间:2019-05-27

Effects of repeated transplantation of human umbilical cord mesenchymal stem cells in different time windows on silica-induced pulmonary fibrosis in rats
Abstract: Objective To investigate the effect of repeated transplantation of human umbilical cord mesenchymal stem cells (HUCMSCs) at different time windows on pulmonary fibrosis in rats exposed to silica dust. Methods Totally 48 healthy male SD rats were randomly divided into 4 groups (n=12), namely, control group, silicosis model group, early treatment group and late treatment group. Non-exposed endotracheal intubation was used to expose rats to 0.5 mL saline (control group) or 0.5 mL silica dust suspension(50 mg/mL mass concentration, the other groups)and repeated one time after a week. Furthermore, 0.5 mL HUCMSCs suspension (cell density 3.0×109/L)were given by tail vein infusion at day 1, day 7, day 14, day 21 in the early treatment group, and at day 28, day 35, day 42, day 49 in the late treatment group after the first dust exposure. Rats in each group were euthanized on the 70 th day of the experiments. The pathological changes of lung tissues were observed by HE staining and Mason staining; the pulmonary fibrosis scores (Ashcroft scores) were marked; the lung coefficients were measured; the levels of lung tissue hydroxyproline(Hyp), transforming growth factor-beta 1(TGF-beta 1) and serum interleukin-6(IL-6) were measured. Results Lung histopathology showed that the alveolar structure was normal in the control group, and fibrosis was observed in the other exposed groups, and cellular fibrous granuloma was observed in silicosis model group. Compared with silicosis model group, collagen fibers in both early and late treatment groups were significantly reduced. Compared with control group, the Aschcroft scores, lung coefficients, lung tissue Hyp, TGF-beta 1 and serum IL-6 in dust-exposed groups were significantly higher (P < 0.01), while those in the both early and late treatment groups were significantly lower than silicosis model group(P < 0.01). There were no significant differences of those indexes between early treatment group and late treatment group (P > 0.05). Conclusion Repeated injection of HUCMSCs, whether in the stage of alveolitis or fibrosis predominated stage, could effectively alleviate silica-induced pulmonary fibrosis in rats.
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