同种异体骨髓间充质干细胞移植对大鼠肝脏缺血-再灌注损伤的影响  

Transplantation of bone marrow mesenchymal stem cells alleviates hepatic ischemia-reperfusion injury in rats

作　　者：郑盛[1] 杨涓[1] 张帆[1] 王玉波[1] 

机构地区：[1]云南省第三人民医院消化内科,云南省昆明市650011

出　　处：《世界华人消化杂志》2016年第11期1639-1648,共10页World Chinese Journal of Digestology

基　　金：云南省自然科学基金资助项目;No.2012FD095;云南省教育厅科研基金重点资助项目;Nos.2014Z125;2015Z146~~

摘　　要：目的：观察同种异体骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs）移植对大鼠肝脏缺血-再灌注损伤（hepatic ischemia reperfusion injury,HIRI）的修复作用并对其可能的治疗机制进行探讨.方法：将健康雌性清洁级SD大鼠的股骨骨髓进行分离、培养、鉴定获得BMSCs;选取60只SD大鼠建立大鼠HIRI模型.动物模型建立后,将90只SD大鼠随机分为BMSCs移植组（n=30,予以尾静脉注射BMSC悬液1 mL,1.0×10~7/mL）,HIRI组（n=30,予以尾静脉注射L-DMEM 1 mL）,另设空白对照组（n=30,予以尾静脉注射生理盐水1 mL）.分别于移植后的1、2、3 wk于各组随机选取5只大鼠取血检测丙氨酸转氨酶（alanine transanminase,ALT）、天冬氨酸转氨酶（aspartate transaminase,AST）、超氧化物歧化酶（superoxide dismutase,SOD）、丙二醛（malondialdehyde,MDA）、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白介素-18（interleukin-18,IL-18）水平;移植后2 wk对肝组织行HE染色,观察移植后肝组织的形态学变化;利用RT-PCR法及Western b l o t法检测肝细胞生长因子（hepatocyte growth factor,HGF）及α-平滑肌蛋白（alphasooth muscle actin,α-SMA）在肝组织中的表达水平.结果：移植后1 wk,BMSCs移植组、HIRI组的血清ALT、AST、MDA、TNF-α和IL-18水平明显高于对照组,SOD活性明显低于对照组,差异均有统计学意义（均P〈0.05）;移植后2、3 wk,BMSCs移植组ALT、AST、MDA、TNF-α和IL-18水平与HIRI组比较明显下调,SOD活性明显升高（均P〈0.05）,但与对照组相比SOD活性仍偏低（P〈0.05）.HE染色后发现,BMSCs移植组大鼠肝细胞变性、坏死及纤维化程度较HIRI组均有明显减轻（P〈0.05）.RT-PCR法检测得知,与HIRI组比较,BMSCs移植组中HGF基因表达水平明显升高,而α-SMA的基因表达水平明显下降（均P〈0.05）.Western blot法检测得知,与HIRI组比较,BMSCs移植组中HGF蛋白表达水平明显升高,而α-SMA的蛋白�AIM： To assess the therapeutic effects of transplantation of bone marrow mesenchymal stem cells（BMSCs） on hepatic ischemiareperfusion injury（HIRI） in rats and explore the underlying mechanism.METHODS： BMSCs were isolated from bone tissues of SD rats, cultured, and identified. SD rats were randomly divided into three groups：a control group, an HIRI group and a BMSCs transplantation group. HIRI was induced by the pringle occlusion method. After hepatic ischemia-reperfusion injury induction, blood samples were taken at 1, 2, and 3 weeks.Serum alanine transaminase（ALT）, aspartate transaminase（AST）, malondialdehyde（MDA）,superoxide dismutase（SOD）, interleukin（IL）-18 and tumor necrosis factor-α（TNF-α）were determined. Liver pathological changes were assessed by HE staining after 2 wk.Expression of hepatocyte growth factor（HGF）and alpha-smooth muscle protein（α-SMA） in the liver was detected by RT-PCR and Western blot.RESULTS： At 1 wk, the levels of serumALT, AST, IL-18, TNF-α and MDA in the transplantation group and HIRI group were significantly higher than those in the control group（P 〈0.05 for all）, and the levels of serum SOD were lower than that in the control group（P 〈0.05）. At 2 wk, the levels of serum ALT, AST, IL-18, TNF-α and MDA in the transplantation group were significantly lower than those in the HIRI group（P〈 0.05 for all）, but the level of serum MDA was higher and the level of serum SOD was lower in the transplantation group than in the control group（P 〈0.05 for both）.Hepatic degeneration, necrosis and fibrosis in the transplantation group were reduced significantly compared with the HIRI group（P〈 0.05）. HGF expression in the liver tissue was significantly higher and α-SMA expression was significantly lower in the transplantation group than in the HIRI group（P〈 0.05 for both）.CONCLUSION： Transplantation of BMSCs can alleviate hepatic ischemia-reperfusion injury via mechanism possibly associated with inhibiting inflam

关 键 词：骨髓间充质干细胞 免疫调节 白介素-18 移植 缺血-再灌注 氧化损伤 

分 类 号：R575[医药卫生—消化系统]