BMSC在I/R扩大肝切除大鼠模型中对肝功能衰竭的防治作用

Prevention and treatment of BM-MSC in hepatic failure in a rat model of I/R expanded hepatectomy

目的探讨缺血再灌注(I/R)扩大肝脏切除大鼠采用骨髓间充质干细胞(BMSC)预处理对大鼠肝功能的影响。方法选取健康雄性SD大鼠54只,采用随机数字表法分为假手术组(假手术处理)、实验组A(等量磷酸缓冲盐预处理后Pringle法肝门阻断30 min,实施肝脏切除手术)、实验组B(BMSC预处理后Pringle法肝门阻断30 min,实施肝脏切除手术)各18只;对比3组大鼠术后6 h、12 h、24 h的血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、直接胆红素(DBIL),采用实时定量PCR技术检测3组大鼠术后24 h肝脏组织中的微小RNA155(miR-155)、肿瘤坏死因子-α(TNF-α)的表达,采用Western-blot方法检测各组大鼠术后24 h肝脏组织中的生长因子样模体黏液样激素样受体(F4/80+)蛋白、Ki67蛋白表达水平;并对3组大鼠术后24 h的肝脏组织进行肝脏缺血再灌注损伤Suzuki评分。结果术后24 h,对各组大鼠肝脏组织实施HE染色后进行病理学观察,可见实验组A和实验组B的Suzuki评分均显著高于假手术组(P<0.05),实验组B的Suzuki评分均显著低于实验组A(P<0.05);术后6、12、24 h实验组A和实验组B的血清ALT、AST、TBIL、DBIL水平均显著高于假手术组(P<0.05),实验组B的血清ALT、AST、TBIL、DBIL水平均显著低于实验组A(P<0.05);术后24 h,实验组A和实验组B的肝组织中miR-155、TNF-αmRNA的表达均显著高于假手术组(P<0.05),实验组B的肝组织中miR-155、TNF-αmRNA的表达均显著低于实验组A(P<0.05);术后24 h实验组A和实验组B的肝组织中Ki67蛋白相对表达强度高于假手术组(P<0.05),F4/80+蛋白相对表达强度低于假手术组(P<0.05);实验组B的肝组织中Ki67蛋白相对表达强度高于实验组A(P<0.05),F4/80+蛋白阳性相对表达强度低于实验组A(P<0.05)。结论I/R扩大肝脏切除大鼠采用BMSC预处理对肝功能具有一定的保护作用,可防治术后发生肝衰竭。

Objective To investigate the effect of preconditioning with bone marrow mesenchymal stem cells(BMSCs)on hepatic function in rats with ischemia-reperfusion(I/R)expansion.Methods A total of 54 healthy male Sprague-Dawley rats were randomly divided into sham operation group(sham operation),experimental group A(liver resection was performed after Pringle method was used for 30 min),Experimental group B(BMSC pretreatment with Pringle method and liver resection),18 rats in each group;serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),direct bilirubin(DBIL)were compared at 6 h,12 h and 24 h after operation in three groups,real-time quantitative PCR was used to detect microRNA155(miR-155),tumor necrosis factor-α(TNF-α)in liver tissue of rats in the three groups after 24 h,Western-blot method was used to detect growth factor-like mucin-like hormone-like receptors F4/80+Protein and Ki67 protein expression levels in liver tissue of rats in each group 24 h after operation;Suzuki scores of liver ischemia-reperfusion injury were detected in liver tissues of the three groups after 24 h.Results HE staining was performed on the liver tissues of each group for pathological observation 24 h after operation.It was found that the Suzuki scores of experimental group A and experimental group B were significantly higher than those of sham operation group(P<0.05),and Suzuki score of experimental group B was lower than experimental group A.The levels of serum ALT,AST,TBIL and DBIL in experimental group A and experimental group B were significantly higher than those in sham operation group at 6 h,12 h and 24 h after operation(P<0.05).The serum levels of ALT,AST,TBIL and DBIL in experimental group B were significantly lower than those in experimental group A(P<0.05).MiR-155 and TNF-αmRNA in liver tissues of experimental group A and experimental group B 24 h after operation were lower than those in sham operation group.The expression of miR-155 and TNF-αmRNA in the liver tissue of experimental group B 24 h after operation was significantly lower than that in experimental group A(P<0.05).The relative expression of Ki67 protein in liver tissue of experimental group B and experimental group A was higher than that in sham operation group(P<0.05),and the relative expression intensity of F4/80+protein was lower than that in sham operation group(P<0.05).The relative expression intensity of Ki67 protein in liver tissue of experimental group B was higher than that of experimental group A(P<0.05),and the relative expression intensity of F4/80+protein was lower than that of experimental group A(P<0.05).Conclusion I/R expansion of liver resection rats with BMSC pretreatment has a certain protective effect on liver function,It can preven and treat liver failure after operation.