中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
Chinese Journal of Emergency Medicine
2015年
9期
994-999
,共6页
徐道剑%张琳%黎俊%张静%何岱昆%钟志越%申捷
徐道劍%張琳%黎俊%張靜%何岱昆%鐘誌越%申捷
서도검%장림%려준%장정%하대곤%종지월%신첩
急性肺损伤%内毒素%间充质干细胞%Wnt/β-Catenin
急性肺損傷%內毒素%間充質榦細胞%Wnt/β-Catenin
급성폐손상%내독소%간충질간세포%Wnt/β-Catenin
Acute lung injury%Endotoxin%Mesenchymal stem cells%Wnt/β-Catenin
目的 探讨Wnt/β-Catenin信号通路在骨髓间充质干细胞(MSCs)治疗急性肺损伤中的作用.方法 采用6只SPF雄性进行贴壁分离法分离培养骨髓间充质干细胞.采用细菌脂多糖诱(LPS)导急性肺损伤,72只6周龄SPF级雄性SD大鼠,体质量约180～220 g,随机(随机数字法)分为4组,分别为:Control组(n=18)采用PBS处理;LPS组(n=18)采用细菌脂多糖诱导急性肺损伤;LPS+ MSCs组(n=18)在进行急性肺损伤的诱导后直接移植MSCs;Control+ MSCs组(n=18)在注射PBS后再移植MSCs.每组分别取LPS注射后6、24和48 h三个时间点.通过肺组织免疫组化分析及免疫印迹分析法检测Wnt信号通路成分的水平.实时定量PCR检测Wnt信号通路的靶基因的表达水平.结果 相比于PBS对照组,移植MSCs的能够显著降低急性肺损伤大鼠的湿干比,同时能显著提高急性肺损伤大鼠的血氧分压的水平.通过组织免疫组化分析,发现Wnt5a在急性肺损伤的肺组织中表达水平有显著的升高,移植的MSCs能够显著降低Wnt5a的水平.免疫印迹结果显示MSCs能够通过降低GSK-3β的磷酸化和β-catenin水平起到治疗的作用.通过实时定量PCR的结果显示MSCs处理能够显著降低Wnt信号靶基因Vegf、Axin2及Klf4的表达.结论 MSCs对急性肺损伤的治疗作用是通过影响Wnt5a的水平,降低GSK-3β和β-catenin的磷酸化及提高Wnt信号靶基因的表达得以发挥的,Wnt信号通路参与到MSCs治疗急性肺损伤的过程中.
目的 探討Wnt/β-Catenin信號通路在骨髓間充質榦細胞(MSCs)治療急性肺損傷中的作用.方法 採用6隻SPF雄性進行貼壁分離法分離培養骨髓間充質榦細胞.採用細菌脂多糖誘(LPS)導急性肺損傷,72隻6週齡SPF級雄性SD大鼠,體質量約180～220 g,隨機(隨機數字法)分為4組,分彆為:Control組(n=18)採用PBS處理;LPS組(n=18)採用細菌脂多糖誘導急性肺損傷;LPS+ MSCs組(n=18)在進行急性肺損傷的誘導後直接移植MSCs;Control+ MSCs組(n=18)在註射PBS後再移植MSCs.每組分彆取LPS註射後6、24和48 h三箇時間點.通過肺組織免疫組化分析及免疫印跡分析法檢測Wnt信號通路成分的水平.實時定量PCR檢測Wnt信號通路的靶基因的錶達水平.結果 相比于PBS對照組,移植MSCs的能夠顯著降低急性肺損傷大鼠的濕榦比,同時能顯著提高急性肺損傷大鼠的血氧分壓的水平.通過組織免疫組化分析,髮現Wnt5a在急性肺損傷的肺組織中錶達水平有顯著的升高,移植的MSCs能夠顯著降低Wnt5a的水平.免疫印跡結果顯示MSCs能夠通過降低GSK-3β的燐痠化和β-catenin水平起到治療的作用.通過實時定量PCR的結果顯示MSCs處理能夠顯著降低Wnt信號靶基因Vegf、Axin2及Klf4的錶達.結論 MSCs對急性肺損傷的治療作用是通過影響Wnt5a的水平,降低GSK-3β和β-catenin的燐痠化及提高Wnt信號靶基因的錶達得以髮揮的,Wnt信號通路參與到MSCs治療急性肺損傷的過程中.
목적 탐토Wnt/β-Catenin신호통로재골수간충질간세포(MSCs)치료급성폐손상중적작용.방법 채용6지SPF웅성진행첩벽분리법분리배양골수간충질간세포.채용세균지다당유(LPS)도급성폐손상,72지6주령SPF급웅성SD대서,체질량약180～220 g,수궤(수궤수자법)분위4조,분별위:Control조(n=18)채용PBS처리;LPS조(n=18)채용세균지다당유도급성폐손상;LPS+ MSCs조(n=18)재진행급성폐손상적유도후직접이식MSCs;Control+ MSCs조(n=18)재주사PBS후재이식MSCs.매조분별취LPS주사후6、24화48 h삼개시간점.통과폐조직면역조화분석급면역인적분석법검측Wnt신호통로성분적수평.실시정량PCR검측Wnt신호통로적파기인적표체수평.결과 상비우PBS대조조,이식MSCs적능구현저강저급성폐손상대서적습간비,동시능현저제고급성폐손상대서적혈양분압적수평.통과조직면역조화분석,발현Wnt5a재급성폐손상적폐조직중표체수평유현저적승고,이식적MSCs능구현저강저Wnt5a적수평.면역인적결과현시MSCs능구통과강저GSK-3β적린산화화β-catenin수평기도치료적작용.통과실시정량PCR적결과현시MSCs처리능구현저강저Wnt신호파기인Vegf、Axin2급Klf4적표체.결론 MSCs대급성폐손상적치료작용시통과영향Wnt5a적수평,강저GSK-3β화β-catenin적린산화급제고Wnt신호파기인적표체득이발휘적,Wnt신호통로삼여도MSCs치료급성폐손상적과정중.
Objective To investigate the role of Wnt/β-Catenin signaling pathway in the endotoxin induced acute lung injury (ALI) during the treatment by mesenchymal stem cells (MSCs).Methods Six SPF male SD rats were isolated and bone marrow mesenchymal stem cells were cultured.A total of 72 SPF male SD rats with 6-week-old were randomly (random number) divided into 4 groups:control group (n =18) in which phosphate buffered solution (PBS) used instead of lipopolysaccharide (LPS);LPS group (n =18) in which LPS used to induce acute lung injury;LPS + MSCs group (n =18) in which MSCs directly transplanted after injection of LPS;Control + MSCs group (n =18) in which MSCs transplanted after injection of PBS.And then 6 rats of each group were sacrificed at 6 h,24 h,and 48 h separately after injection of LPS.At 24 h after the modeling,lung tissue was taken and the levels of Wrnt signaling pathway components were detected by using immunohistochemistry and Western blot.In addition,quantitative realtime PCR was used to detect the expression of Wnt signaling pathway target genes.Results Compare with the PBS control group,significant decrease in lung dry-to-wet ratio and increase in arterial oxygen partial pressure (PaO2) were found in MSCS transplantation groups.According the immunohistological results,Wnt 5a was significantly increased in the LPS-induced ALI rats and decreased after MSCs transplantation.Moreover,decrease in levels of GSK-3β phosphorylation and β-catenin was found in the lung tissue after MSCs transplantation.In addition,the expressions of Wnt signaling target genes Vegf,Axin2 and Klf4 were decreased significantly after MSCs transplantation.Conclusions In the setting of ALI,the therapeutic effect of MSCs was exerted by decreasing the expressions of Wnt 5a,GSK-3β phosphorylation,β-catenin,and Wnt signaling target genes Vegf,Axin2 and Klf4.Wnt signaling implicated in the therapeutic effect of MSC in the setting of ALI.
