摘要
目的:研究外源性骨髓间充质干细胞(BMSCs)移植对大鼠矽肺纤维化的疗效,并探讨其量效关系。方法:体外分离、培养雄性5周龄SD大鼠BMSCs。将50只健康SD雌性大鼠随机分为5组:对照组、矽肺模型组、BMSCs治疗A组(1×109/L)、BMSCs治疗B组(3×109/L)和BMSCs治疗C组(5×109/L),每组10只。采用非暴露式气管插管一次性灌注二氧化硅(SiO2)粉尘混悬液法建立大鼠矽肺模型,并给予不同剂量BMSCs干预治疗,各组大鼠于21 d处死取材。采用HE染色和Masson染色观察肺组织形态学变化;免疫组织化学法检测肿瘤坏死因子α(TNF-α)和转化生长因子β(TGF-β)的定位和分布;免疫印迹法检测TNF-α、TGF-β、Ⅰ型胶原和Ⅲ型胶原的蛋白表达量;免疫荧光法检测性别决定区Y(SRY)蛋白证实BMSCs的归巢。结果:矽肺模型组较对照组大鼠有明显炎症细胞浸润、矽结节形成、胶原沉积等病理改变,BMSCs治疗A组较矽肺模型组病理改变有所缓解,BMSCs治疗B组病情缓解更加显著,但BMSCs治疗C组较矽肺模型组病理改变加重。大鼠肺组织TNF-α、TGF-β、Ⅰ型胶原和Ⅲ型胶原表达的情况:BMSCs治疗C组>矽肺模型组>BMSCs治疗A组>BMSCs治疗B组>对照组,除BMSCs治疗C组与矽肺模型组比较差异无统计学意义外(P>0.05),其它各组间比较差异均有统计学意义(P<0.05)。激光扫描共聚焦显微镜观察到BMSCs治疗B组大鼠肺组织中SRY阳性表达,心、肝、脾、肾组织中未见明显表达。结论:外源性BMSCs移植对大鼠矽肺纤维化有一定的拮抗作用,并且存在剂量-效应关系。
AIM： To study the effects of exogenous bone mesenchymal stem cell （BMSC） transplantation on silicosis fibrosis in rats, and to explore the dose - effect relationship. METHODS ： BMSCs were isolated and cultured from male 5 - week - old SD rats in vitro. Fifty healthy female SD rats were randomly divided into 5 groups ： control group, sili- eosis model group, BMSCs treatment A group （ 1×109 cells/L）, BMSCs treatment B group （3×109cells/L） and BMSCs treatment C group （5×109 cells/L）. The silicosis model was made by one -time infusion of silica dust suspension using the non -exposed tracheal intubation, and different doses of BMSCs were given for intervention therapy. All the rats were sacrificed on the 21st day after the model was established. The morphological changes of the lung tissues were observed by HE staining and Masson staining. The localization and distribution of tumor necrosis factor α （TNF -α） and transforming growth factor β （ TGF - β） were determined by the method of immunohistochemistry. The protein levels of TNF -α, TGF -β, collagen type Ⅰ and collagen type Ⅲ were detected by Western blotting. The sex -determining region （SRY） protein was searched by an immunofluorescence method to confirm the homing of BMSCs. RESULTS： Compared with control group, the silicosis model group had significant alveolitis changes, silicon nodule formation, collagen deposition and other pathological characteristics. Compared with silicosis model group, the pathological changes in BMSCs treatment A group were improved. The conditions of BMSCs treatment B group were also improved significantly. However, the pathological changes in BMSCs treatment C group were increased obviously. The protein levels of TNF - α, TGF - β, collagen type Ⅰ and collagen type Ⅲ in the lung tissues ranked as follows： BMSCs treatment C group 〉 silicosis model group 〉 BMSCs treatment A group 〉 BMSCs treatment B group 〉 control group. The difference between BMSCs treatment C group and sil- icosis model group was not statistically significant, and the differences between the other groups were statistically signifi- cant. The SRY -positive cells were observed in BMSCs treatment B group, but no significant expression in the heart, liv- er, spleen and kidney tissues was observed. CONCLUSION： The exogenous BMSC transplantation antagonizes the devel- opment of silicosis fibrosis in rats, which has dose - effect relationship.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第12期2205-2210,共6页
Chinese Journal of Pathophysiology
基金
河北省科技支撑重点专项基金资助项目(No.09276191D)
国家安全生产监督管理局科技发展计划资助项目(No.06-549)
关键词
矽肺
骨髓间充质干细胞
Ⅰ型胶原
Ⅲ型胶原
肿瘤坏死因子α
转化生长因子Β
Silicosis
Bone mesenchymal stem cells
Collagen type Ⅰ
Collagen type Ⅲ
Tumor necrosis factor alpha
Transforming growth factor beta
