摘要
目的通过原位气管移植建立闭塞性细支气管炎(OB)模型,探讨骨髓间充质干细胞(BMSCs)对OB的治疗效果。方法Brown Norway→Lewis(BN→Lew)大鼠气管原位移植建模,术后分别给予15 mg/(kg·d)环孢菌素A(CsA)灌胃至预杀期,BMSCs单细胞混悬液1 ml(1×106/ml),生理盐水(NS)1 ml尾静脉注射治疗。移植后第7、30和60天获取标本,组织切片,苏木素-伊红(HE)染色,行组织病理观察,测量管腔阻塞率和上皮细胞脱落率;流式细胞术检测外周血CD4+、CD8+T细胞比例;Western blot法检测移植物转化生长因子-β1(TGF-β1)的表达。结果NS、CsA和BMSCs组移植术后60 d气道阻塞率分别为(52.62±10.21)%、(28.52±8.81)%和(21.30±2.27)%,CsA和BMSCs显著减轻气道阻塞(P=0.030,P=0.004);气道上皮脱落率分别为(96.10±0.86)%、(93.67±4.04)%和(25.26±1.10)%,BMSCs组较NS组和CsA组低(P=0.001,P=0.009);60 d外周血CD8+T细胞比例分别为(24.13±1.82)%、(49.50±3.03)%和(26.27±0.40)%,BMSCs组较NS组和CsA组低有效抑制外周血CD8+T细胞增殖(P=0.007);60 d TGF-β1相对表达量分别为1.21、0.77和0.36,BMSCs组表达较NS组和CsA组低(P=0.000)。结论在大鼠原位气管移植OB模型中,BMSCs可下调TGF-β1表达,减轻CD8+T细胞浸润,保护上皮细胞,从而明显减轻气道阻塞。
Objective Through orthotopic tracheal transplantation model to analyze the therapeutic effect of bone marrow mesenchymal stem cells (BMSCs) on obliterative bronchiolitis (OB).Methods Brown Norway-to-Lewis rats were subjected to orthotopic tracheal transplantation, and intragastically given 15 mg/(kg·d) cyclosporine (CsA) until the day when the animals were ready to be killed. BMSCs single cell suspension liquid [1 ml, (1×106/ml) and normal saline (NS) (1 ml) were injected via the tail vein, respectively. Tracheal grafts were harvested at 7th, 30th, and 60th days post-transplantation. The tracheal luminal obliteration rate and epithelial loss were assessed on sections stained with hematoxylin and eosin. CD4+ and CD8+ T lymphocytes in peripheral blood were aexamined using flow cytometry. The expression of transforming factor-beta1 (TGF-β1) was detected by Western blotting.Results The luminal obliteration in NS, CsA and BMSCs treated groups after 60 days post-transplant were (52.62±10.21)%, (28.52±8.81)% and (21.30±2.27)% respectively, CsA and BMSCs were significantly attenuated (P=0.030, P=0.004); the epithelial loss were (96.10±0.86)%, (93.67±4.04)% and (25.26±1.10)% respectively, BMSCs were protective against the epithelial loss post-transplant compared with NS and CsA group (P=0.009); the percentage of CD8+ T lymphocyte in peripheral blood were (24.13±1.82)%, (49.50±3.03)% and (26.27±0.40)% respectively, which were significantly suppressed by BMSCs compared with NS and CsA group (P=0.007). The relative expression quantity of TGF-β1 were 1.21, 0.77 and 0.36 respectively, which can decrease TGF-β1 expression induced by transplant (P=0.000).Conclusion BMSCs could prevent development of airway occlusion through downregulating the expression of TGF-β1, reducing the CD8+ T lymphocyte infiltration and enhancing epithelial preservation in a orthotopic tracheal transplantation mouse model of OB.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2017年第1期75-78,共4页
Chinese Journal of Experimental Surgery
基金
福建省自然科学基金(2012J01354)
关键词
原位气管移植
骨髓间充质干细胞
环孢素
肺移植
闭塞性细支气管炎
大鼠
Orthotopic tracheal transplantation
Bone marrow mesenchymal stem cells
Cyclosporine
Lung transplantation
Obliterative bronchiolitis
Rat