骨髓基质细胞在脂多糖诱导的肺损伤小鼠肺组织的定植研究

Engraftment of bone marrow stromal cells in lipopolysaccharide-injured mouse lungs

摘要:

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14 d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（AEC II）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）-5 mRNA的表达。结果：移植后14 d，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AEC II 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±0.18 vs 1.38±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。［中国当代儿科杂志，2009，11（5）：321-327］

Abstract:

OBJECTIVE: To explore a feasibility of engraftment of systemically transplanted bone marrow stromal cells (BMSCs) and differentiation into lung epithelial cells in lipopolysaccharides (LPS)-injured lungs. METHODS: BMSCs were isolated from bone marrow of transgenic green fluorescent protein (GFP) C57BL/6J mice and systemically administered to bone marrow-suppressed wild-type C57BL/6J mice. A mouse model of lung injury was prepared by intratracheal instillation of LPS. Recipients were assigned to four groups: intratracheal PBS + BMSCs transplantation (CM), intratracheal LPS + BMSCs transplantation (LM), intratracheal PBS + irradiation + BMSCs transplantation (CIM) and intratracheal LPS+ irradiation + BMSCs transplantation (LIM). BMSCs engraftment in recipient lungs was determined by immunofluorescent staining 14 days after BMSCs administration. Alveolar epithelial type II cells were isolated from recipient lungs and the rate of GFP positive cells was measured by flow cytometry. Expression of surfactant protein (SP)-A, SP-C and aquaporin (AQP)-5 mRNA in the lungs was evaluated by real-time PCR. RESULTS: GFP and cytokeratin positive cells were observed in lung parenchyma of the CIM and the LIM groups, but not in the CM and the LM groups. The LIM group had more positive cells than the CIM group. The rates of GFP positive cells were higher in the CIM (11.10±3.19%) and the LIM groups (14.40±2.40%) than those in the CM and the LM groups (2.82±1.03% and 3.81±0.93%, respectively; P<0.05). The LIM group had higher mRNA expression of SP-C than the CM group (2.09±0.18 vs 1.38±0.30; P<0.05). CONCLUSIONS: Donor derived BMSCs can engraft in LPS-injured lungs and differentiate into lung epithelial cells, suggesting BMSCs transplantation might contribute to lung repair.［Chin J Contemp Pediatr, 2009, 11 (5):321-327］