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目的探讨静脉输注骨髓基质细胞(BMSCs)在脂多糖(LPS)诱导的肺损伤小鼠肺组织定植并向肺泡上皮细胞分化的可能性。方法绿色荧光蛋白(GFP)转基因C57BL/6J小鼠作为BMSCs移植供体,同种野生型小鼠为移植受体。气道滴入LPS诱导肺损伤。受体分为以下几组:(1)PBS+BMSCs移植(CM);(2)LPS+BMSCs移植(LM);(3)PBS+全身放射+BMSCs移植(CIM);(4)LPS+全身放射+BMSCs移植(LIM)。移植14d后,以免疫荧光双标染色检测BMSCs在受体肺组织的定植情况,流式细胞仪检测体外培养的肺泡II型上皮细胞(AECII)GFP阳性率,荧光定量PCR法检测肺组织表面活性物质蛋白(SP)-A、SP-C和水通道蛋白(AQP)-5mRNA的表达。结果移植后14d,免疫荧光双标染色可见CIM和LIM组有少量肺泡上皮细胞呈GFP和角蛋白双染阳性,而且LIM组较CIM组有较多阳性细胞。与CM和LM组(分别为2.82±1.03%和3.81±0.93%)比较,CIM和LIM组的AECII GFP阳性率更高(分别为11.10±3.19%和14.40±2.40%;P<0.05)。LIM组SP-C mRNA表达较CM组增高(2.09±0.18vs1.38±0.30;P<0.05)。结论供体来源的BMSCs能在LPS诱导的受损肺组织定植分化,提示静脉输注BMSCs可能有助于肺损伤的修复。
Objective To investigate the possibility of intravenous infusion of bone marrow stromal cells (BMSCs) in the lung tissue of mice with lung injury induced by lipopolysaccharide (LPS) and their differentiation into alveolar epithelial cells. Methods Green fluorescent protein (GFP) transgenic C57BL / 6J mice were used as donor for BMSCs transplantation and allogeneic wild type mice were used as transplant recipients. Instillation of airway LPS induces lung injury. The recipients were divided into the following groups: (1) PBS + BMSCs transplantation (CM); (2) LPS + BMSCs transplantation (LM); (3) PBS + systemic irradiation + BMSCs transplantation BMSCs transplantation (LIM). Fourteen days after transplantation, the colonization of BMSCs in recipient lung tissue was detected by double immunofluorescence staining. The positive rate of GFP in cultured alveolar type II epithelial cells (AECII) was detected by flow cytometry. The surface activity of lung tissue was detected by real-time fluorescence quantitative PCR Substance Protein (SP) -A, SP-C, and Aquaporin (AQP) -5 mRNA expression. Results 14 days after transplantation, immunofluorescence double-stained staining showed that a small amount of alveolar epithelial cells in CIM and LIM groups were positive for GFP and keratin, and there were more positive cells in LIM group than CIM group. Compared with CM and LM groups (2.82 ± 1.03% and 3.81 ± 0.93%, respectively), AECII GFP positive rates were higher in CIM and LIM groups (11.10 ± 3.19% and 14.40 ± 2.40%, respectively; P <0.05). The expression of SP-C mRNA in LIM group was higher than that in CM group (2.09 ± 0.18vs1.38 ± 0.30; P <0.05). Conclusion Donor-derived BMSCs can colonize and differentiate into damaged LPS-induced lung tissue, suggesting that intravenous infusion of BMSCs may contribute to the repair of lung injury.