摘要
目的探讨同种异体骨髓间充质干细胞(BM-MSCs)对大鼠肝脏热缺血再灌注损伤的保护作用。方法取10只健康Wistar大鼠骨髓,体外分离、培养间充质干细胞,对其进行生物学鉴定,并对门静脉途径移植入肝脏的间充质干细胞行4,6-联脒-2-苯基吲哚(DAPI)染色示踪。建立大鼠70%肝脏缺血再灌注损伤模型,将32只雄性Wistar大鼠随机分成假手术组(Sham组)、缺血再灌注组(I/R组)、维生素C治疗组(VC组)及BM-MSCs组,每组8只,分别于再灌注24 h后取材,检测血清天门冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)水平,肝脏组织总超氧化物歧化酶(SOD)和丙二醛(MDA)含量,并对肝脏组织行HE染色观察病理学改变,TUNEL法检测肝脏凋亡指数(AI)。结果体外分离的BM-MSCs生长稳定,增殖旺盛,表达CD29及CD44,不表达CD34及CD45;大鼠肝脏缺血再灌注损伤模型稳定,在肝脏组织切片内见间充质干细胞定植,多位于肝小叶中央静脉周围。VC组和BM-MSCs组的AST、ALT、MDA和AI均较I/R组低(P<0.05),SOD均较I/R组高(P<0.05);BM-MSCs组的AST、ALT、MDA和AI均低于VC组(P<0.05),SOD高于VC组(P<0.05)。结论 BM-MSCs可通过减轻氧化应激损伤和抑制肝细胞凋亡,来保护大鼠肝脏热缺血再灌注损伤。
Objective To explore repair role of allogeneic bone marrow mesenchymal stem cells（BM-MSCs） transplantation on treating hepatic ischemia reperfusion injury（HIRI） in rats.Methods Ten rats were executed to get BM-MSCs,then BM-MSCs were cultured in vitro and dyed by 4,6-diamidino-2-phenylindole（DAPI）.Models of 70% hepatic ischemia reperfusion injury were eatablished.Thirty two rats were randomly divided into sham operation group（Sham group）,ischemia reperfusion group（I/R group）,Vitamin C group（VC group）,and BM-MSCs group.Serum samples were analyzed for ALT and AST,and hepatic tissue were for superoxide dismutase（SOD） and malondialdehyde（MDA）.Liver sections were stain with hematoxylin and eosin（HE） for histological analysis,TUNEL staining was applied to detect hepatic apoptosis.Serum and tissues were both collected at 24 h after reperfusion.Results The isolated BM-MSCs maintained vigorous growth in vitro.Specific markers for MSCs antigens CD29 and CD44 were detected by flow cytometry,but antigens CD34 and CD45 were not be detected.Models of HIRI were stable,and BM-MSCs were detected around the periportal area by DAPI staining.Compared with I/R group,levels of ALT,AST,MDA,and AI in the VC group and BM-MSCs group decreased at 24 h after reperfusion（P0.05）,meanwhile SOD level increased（P0.05）.Compared with VC group,levels of ALT,AST,MDA,and AI in the BM-MSC group decreased at 24 h after reperfusion（P0.05）,meanwhile SOD level increased（P0.05）.Conclusion BM-MSCs could protect HIRI by alleviating oxidative stress and inhibiting cellular apoptosis.
出处
《中国普外基础与临床杂志》
CAS
2013年第1期60-65,共6页
Chinese Journal of Bases and Clinics In General Surgery
关键词
骨髓间充质干细胞
肝脏缺血再灌注损伤
氧化应激
凋亡
干细胞治疗
Bone marrow derived mesenchymal stem cells
Hepatic ischemia reperfusion injury
Oxidative stress
Apoptosis
Stem cell therapy
