CAJ | 학술논문

目的观察静脉移植的骨髓间充质干细胞(MSC)在烟雾吸入性损伤家兔体内"归巢"和分化情况.方法将32只新西兰大耳白兔按照随机数字表法分为正常对照组、吸入性损伤组、正常对照+MSC处理组和MSC治疗组,每组8只.正常对照组家兔不致伤,经耳缘静脉注入10 mLPBS;正常对照+MSC处理组家兔不致伤,经耳缘静脉注入内含1×107个溴脱氧尿苷(BrdU)标记的第3代兔MSC的PBS 10 mL.其余2组家兔造成重度烟雾吸入性损伤后,吸入性损伤组同正常对照组处理,MSC治疗组同正常对照+MSC处理组处理.4组家兔分别于注射后7或28 d,取肺组织、气管组织行免疫组织化学染色观察MSC在受损组织中"归巢"情况;核标记物BrdU分别与肺或气管内特异性膜标记物水通道蛋白5(AQP-5)、碱性磷酸酶(AKP)、CD34以及角蛋白组合,行免疫组织化学双染色,观察MSC向功能细胞分化情况;HE染色观察肺及气管损伤情况.结果 (1)免疫组织化学染色显示,MSC治疗组可见MSC于注射后7 d在肺组织和气管组织中"归巢";正常对照+MSC处理组未见此现象.(2)免疫组织化学双染色显示,MSC治疗组于注射后28 d肺组织中可见AQP-5和AKP以及CD34表达阳性的MSC,气管组织中未见角蛋白表达阳性的MSC,正常对照+MSC处理组未见上述阳性表达细胞.(3)HE染色显示,MSC治疗组肺损伤和气管损伤较吸入性损伤组明显减轻,Fb增殖显著减少.结论静脉移植的MSC能"归巢"至烟雾吸入性肺损伤炎症反应明显的肺组织和气管组织区域,并分化为肺泡Ⅰ型上皮细胞、肺泡Ⅱ型上皮细胞和肺血管内皮细胞,可能参与了烟雾吸入性损伤的组织修复过程.
목적 관찰정맥이식적골수간충질간세포(MSC)재연무흡입성손상가토체내"귀소"화분화정황.방법장32지신서란대이백토안조수궤수자표법분위정상대조조、흡입성손상조、정상대조+MSC처리조화MSC치료조,매조8지.정상대조조가토불치상,경이연정맥주입10 mLPBS;정상대조+MSC처리조가토불치상,경이연정맥주입내함1×107개추탈양뇨감(BrdU)표기적제3대토MSC적PBS 10 mL.기여2조가토조성중도연무흡입성손상후,흡입성손상조동정상대조조처리,MSC치료조동정상대조+MSC처리조처리.4조가토분별우주사후7혹28 d,취폐조직、기관조직행면역조직화학염색관찰MSC재수손조직중"귀소"정황;핵표기물BrdU분별여폐혹기관내특이성막표기물수통도단백5(AQP-5)、감성린산매(AKP)、CD34이급각단백조합,행면역조직화학쌍염색,관찰MSC향공능세포분화정황;HE염색관찰폐급기관손상정황.결과 (1)면역조직화학염색현시,MSC치료조가견MSC우주사후7 d재폐조직화기관조직중"귀소";정상대조+MSC처리조미견차현상.(2)면역조직화학쌍염색현시,MSC치료조우주사후28 d폐조직중가견AQP-5화AKP이급CD34표체양성적MSC,기관조직중미견각단백표체양성적MSC,정상대조+MSC처리조미견상술양성표체세포.(3)HE염색현시,MSC치료조폐손상화기관손상교흡입성손상조명현감경,Fb증식현저감소.결론정맥이식적MSC능"귀소"지연무흡입성폐손상염증반응명현적폐조직화기관조직구역,병분화위폐포Ⅰ형상피세포、폐포Ⅱ형상피세포화폐혈관내피세포,가능삼여료연무흡입성손상적조직수복과정.
Objective To observe the homing and differentiation of marrow-derived mesenchymal stem cells (MSC) transplanted intravenously in smoke inhalation injured rabbits. Methods Thirty-two New Zealand big ear rabbits were divided into normal control group ( NC ), inhalation injury group ( Ⅱ),normal control + MSC treatment group (NM) , and MSC treatment group (MT) according to the random number table, with 8 rabbits in each group. Rabbits in NC group were injected with 10 mL phosphate buffered saline (PBS) via ear marginal vein. Rabbits in NM group were injected with 10 mL PBS containing the third generation MSC labeled by BrdU ( 1 × 107 per 10 mL PBS) via ear marginal vein. Severe smoke inhalation injury model was reproduced in the other two groups, among them rabbits in Ⅱ group were treated as rabbits in NC group, rabbits in MT group treated as rabbits in NM group. On the 7th and 28th day post treatment (PTD), lung tissue and trachea tissue were harvested from four groups for observation on injury with HE staining. Homing of MSC in injured tissue was observed with immunohistochemistry staining. The differentiation of MSC into functional cells was observed with immunohistochemical double staining of combining nuclear marker BrdU with lung (trachea) membrane-specific marker aquaporin-5 (AQP-5) , alkaline phosphatase (AKP), CD34, and cytokeratin respectively. Results (1) MSC homing in lung and trachea tissue was observed in MT group on PTD 7, which was not observed in NM group. (2) AQP-5, AKP, and CD34 positive MSC were observed in lung tissue in MT group on PTD 28, while cytokeratin positive MSC was not observed in trachea tissue. No positively marked MSC was observed in NM group. (3) Injury in lung and trachea was less severe in MT group than in Ⅱ group; and the proliferation of fibroblasts was less in MT group. Conclusions Intravenous injection of MSC to rabbits with smoke inhalation injury can migrate to lung and trachea tissue at obviously inflammatory site, and differentiate into alveolar epithelial cells type Ⅰ and Ⅱ , and pulmonary vascular endothelial cells, which may participate in the process of tissue repair in smoke inhalation injury.