中性粒细胞在骨髓间充质干细胞治疗D-氨基半乳糖诱导急性肝衰竭大鼠中的作用  

作　　者：赵鑫[1] 施晓雷[1] 张志恒[1] 马虎成[1] 袁献温[1] 丁义涛[1] 

机构地区：[1]南京大学医学院附属鼓楼医院,210008

出　　处：《中华肝脏病杂志》2016年第8期601-607,共7页Chinese Journal of Hepatology

基　　金：国家自然科学基金（81170418）;江苏省自然科学基金（BK20131084）

摘　　要：目的 观察骨髓间充质干细胞（MSCs）移植对D-氨基半乳糖（D—GalN）诱导SD大鼠急性肝衰竭模型的治疗作用并探索中性粒细胞在其中的机制关联。方法将39只雄性SD大鼠分为4组：对照组（／7=8，腹腔注射等渗盐水）、模型组n=10，腹腔注射D—GaIN）、溶剂组n=9，腹腔注射D-GalN后2h尾静脉注射等渗盐水）及治疗组（n=12，腹腔注射D—GaIN后2h尾静脉注射MSCs）。建立D-GaIN诱导急性肝衰竭模型后24h处死大鼠，分别收集血液及肝脏组织，检测血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶和总胆红素，血液分析大鼠外周血中性粒细胞数量及比率，免疫荧光检测肝脏中性粒细胞标志物Ly6g表达水平，髓过氧化物酶（MPO）试剂盒检测肝脏MPO活性，RT—PCR检测肝脏组织炎性因子及趋化因子肿瘤坏死因子。a、白细胞介素1β、干扰素Y、白细胞介素10、CXC趋化因子配体1、CXC趋化因子配体2的mRNA水平；另将64只雄性SD大鼠随机分组后连续7d观察各组大鼠生存率。两两组间比较应用独立样本t检验（经Levene方差齐性检验，当JD〈0．05时，采用校正f检验）；大鼠生存分析采用两两比较的Logrank检验。结果D-GaIN诱导的急性肝衰竭SD大鼠24h后反映肝功能的生物化学指标显著升高[丙氨酸氨基转移酶：（2884．1±541．0）U／L与（45．4±11．0）U／L，P〈0．001；天冬氨酸氨基转移酶：（3634．9±755．9）U／L与（143．9±23．7）U／L，P〈0．001；总胆红素：（44．4±8．4）μmmol／L与（0．9±0．2）ummol／L，P〈0．001]外周血中性粒细胞数量与比率大幅上升[数量：（4．7±1．1）×10^9与（1．4±0．4）×10^9，P〈0．001；比率：44．9％±8．0％与18．3％±4．4％，P〈0．001）；肝脏组织中性粒细胞大量聚集，MPO活性[（4．72±1．09）U／g与（1．13±0．24）U／g，P〈0．001]、炎性因子及趋化因子均明显增多。MSCs移植�Objective To investigate the therapeutic effect of bone marrow mesenchymal stromal cell (BMSC) transplantation on D-galactosamine-induced acute liver failure in Sprague-Dawley (SD) rats,as well as the mechanism of neutrophils in this process.Methods A total of 39 male SD rats were divided into control group (8 rats,intraperitoneal injection of isotonic saline),model group (10 rats,intraperitoneal injection of D-galactosamine),solvent group (9 rats,tail vein injection of isotonic saline at 2 hours after intraperitoneal injection of D-galactosamine),and treatment group (12 rats,tail vein injection of MSCs at 2 hours after intraperitoneal injection of D-galactosamine).The rats were sacrificed at 24 hours after the model of D-galactosamine-induced acute liver failure was established,and the blood and liver tissue were harvested.The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and total bilirubin (TBil) were measured,and blood analysis was performed to measure the number and percentage of neutrophils in peripheral blood.Immunofluorescence assay was used to measure the expression of the neutrophil marker Ly6g in the liver,the myeloperoxidase (MPO) kit was used to measure the activity of MPO in liver,and RT-PCR was used to measure the mRNA expression of inflammatory cytokines and chemokines in the liver,i.e.,tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β),interferon-γ (IFN-γ),interleukin-10 (IL-10),CXC chemokine ligand 1 (CXCL1),and CXC chemokine ligand 2 (CXCL2).Another 64 male SD rats were randomly divided into groups,and the survival rates of rats in each group were observed for 7 days.The independent samples t-test was used for comparison between any two groups (Levene homogeneity test of variance,and the corrected t-test was used for a P value of ＜ 0.05),and the log-rank test was used for comparison of survival rates between any two groups.Results At 24 hours after acute liver failure was induced by D-galactosamine in the SD rats,there were significant increases

关 键 词：肝功能衰竭 急性 间充质干细胞 中性粒细胞 

分 类 号：R575.3[医药卫生—消化系统]