摘要
目的比较骨髓间充质干细胞(BMMSCs)经静脉和经气管2种移植方式对染矽尘大鼠的肺损伤修复作用。方法取14只无特定病原体级健康雄性SD大鼠培养BMMSCs,另取48只同类大鼠随机分为对照组、染尘组、静脉组和气管组,每组12只。对照组大鼠气管内注入无菌生理氯化钠溶液1.0 m L,另3组大鼠均气管内注入质量浓度为40 g/L的无菌矽尘混悬液1.0 m L,静脉组、气管组再分别经鼠尾静脉和气管内注入细胞密度为5×109个/L的BMMSCs 0.5 m L。第28天放血处死大鼠,检测肺脏脏器系数;观察肺组织病理学改变情况,进行肺泡炎症和肺间质纤维化评分;检测肺组织羟脯氨酸(HYP)、转化生长因子-β1(TGF-β1)和肿瘤坏死因子-α(TNF-α)水平,以及周围血血常规和血生化指标。结果染尘组、静脉组和气管组大鼠肺脏脏器系数均高于对照组(P<0.05),静脉组和气管组大鼠肺脏脏器系数均低于染尘组(P<0.05),但静脉组和气管组大鼠肺脏脏器系数差异无统计学意义(P>0.05)。肺组织病理学观察显示:对照组大鼠肺组织结构正常;染尘组肺部炎症最严重,并可见大量矽结节和胶原纤维沉积;静脉组和气管组炎症、矽结节数量以及胶原纤维增生均较染尘组有改善。染尘组、静脉组和气管组大鼠的肺泡炎、肺间质纤维化评分以及肺组织TGF-β1、TNF-α水平均高于对照组(P<0.05),静脉组和气管组大鼠上述4个指标均低于染尘组(P<0.05),但静脉组和气管组大鼠上述4个指标差异均无统计学意义(P>0.05)。染尘组大鼠肺组织HYP水平高于其余3组(P<0.05);但静脉组、气管组和对照组HYP水平分别两两比较差异均无统计学意义(P>0.05)。染尘组、静脉组和气管组大鼠周围血白细胞计数、红细胞计数、血红蛋白水平、红细胞压积均分别高于对照组(P<0.05)。气管组大鼠血清尿素氮水平高于静脉组(P<0.05),染尘组大鼠血清铜蓝蛋白水平高于其余3组(P<0.05)。4组大鼠周围血血小板和血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、肌酸激酶、总蛋白、白蛋白、球蛋白、肌酐等指标的差异均无统计学意义(P>0.05)。结论经静脉和经气管2种BMMSCs移植方式对矽尘所致大鼠肺损伤均具有修复作用,2种移植方式的修复作用没有显著性差异。
Objective To compare different effects of intravenous injection and trachea transplantation ways of bone marrow-derived mesenchymal stem cell （BMMSC） on the damage repair of lung injury in rats exposed to silica dust. Methods BMMSCs were isolated from 14 specific pathogen free healthy male SD rats, cultured and amplified in vitro, while 48 rats of the same kind were randomly divided into control group, silica-exposed group, intravenous injection way group, and trachea way group, 12 rats in each group. The latter 3 groups were injected intratracheally with 1.0 mL of 40 g/L sterile silica dust suspension while control group was given 1.0 mL of sterile physiological sodium chloride solution. Then, intravenous injection way group and trachea way group were injected with 0.5 mL of BMMSCs （cell density, 5 ~ 109/L） through the tail intravenous injection and trachea respectively. Rats were weighted and executed on day 28, and lung organ coefficients were calculated. The pathomorphological changes in lung tissues were observed and alveolar inflammation and pulmonary fibrosis were scored, while hydroxyproline （HYP）, transforming growth factor-beta 1 （TGF-β1） and tumor necrosis factor-α （TNF-α） concentration of lung, routine blood test and some biochemical indices of peripheral blood were also detected. Results The lung organ coefficients of silica-exposed group, intravenous injection way group, and trachea way group were higher than that of control group respectively （P 〈 0.05 ）, while the lung organ coefficients of intravenous injection way group and trachea way group were lower than that of silica-exposed group （ P 〈 0. 05）, but there was no significant difference in the lung organ coefficients between intravenous injection way group and trachea way group （ P 〉 0. 05 ）. The pathomorphological observation showed normal lung tissue structure in control group. Lung sections showed that inflammation was the most serious in the silica-exposed group with numerous silicotic nodules and collagen fiber deposition. Pulmonary inflammation, the number of silicotic nodules and collagen fiber deposition of intravenous injection way group and trachea way group were less than those of silica-exposed group. The scores of alveolar inflammation and pulmonary fibrosis and the pulmonary TGF-β1 and TNF-α levels of silica-exposed group, intravenous injection way group, and trachea way group were higher than those of the control group, respectively （ P 〈 0. 05 ）, while the above 4 indices of intravenous injection way group and trachea way group were lower than those of the silica-exposed group （ P 〈 0. 05 ） , but there was no statistical significant difference between intravenous injection way group and trachea way group （P 〉 0.05）. The pulmonary HYP content of silica-exposed group was higher than those of the other 3 groups （P 〈 0. 05）, but the index of HYP content showed no statistical significant difference between any two groups among the intravenous injection way group, trachea way group, and control group （P 〉 0. 05）. The counts of white blood cell and erythrocytes, hemoglobin level, and hematocrit of peripheral blood in silica-exposed group, intravenous injection way group, and trachea way group were higher than those in control group, respectively （ P 〈 0. 05 ）. Serum urea nitrogen concentration in trachea way group was higher than that in intravenous injection way group （P 〈 0. 05）, and the serum ceruloplasmin concentration of silica-exposed group was higher than those of the other 3 groups （ P 〈 0. 05 ）. There were no statistical significant differences in peripheral blood platelet count, serum alanine aminotransferase, aspartate aminotransferase, creatine kinase activities, serum total protein, albumin, globulin, and creatinine concentrations in the 4 groups （P 〉 0. 05）. Conclusion BMMSC transplantation through intravenous injection and trachea ways has the similar effects with no significant difference on the damage repair of lung injury induced by silica dust in rats.
出处
《中国职业医学》
CAS
北大核心
2015年第4期366-371,共6页
China Occupational Medicine
基金
国家科技支撑计划(2014BAI12B01)
国家自然科学基金(81302396)
国家临床重点专科建设项目(2011-09)
广东省职业病防治重点实验室(2012A061400007)
广东省医学科研基金(A2015233)
关键词
骨髓间充质干细胞
移植方式
二氧化硅
矽肺
肺损伤
修复
大鼠
Bone marrow-derived mesenchymal stem cells
Transplantation way
Silica
Silicosis
Lung injury
Repair
Rat
