肝脏自然杀伤细胞对梗阻性黄疸小鼠模型肝纤维化的影响  

Effect of liver natural killer cells on liver fibrosis in mice with obstructive jaundice

作　　者：刘传江[1] 周文婧[2] 许先玲[1] 秦涛[1] 陈琳[1] 付强[1] 胡明星[1] 王玉柱[1] 

机构地区：[1]河南省人民医院肝胆胰腺外科郑州大学人民医院,450003 [2]河南省人民医院内分泌科郑州大学人民医院,450003

出　　处：《中华实验外科杂志》2017年第3期406-409,共4页Chinese Journal of Experimental Surgery

摘　　要：目的 观察肝脏自然杀伤（NK）细胞对梗阻性黄疸小鼠模型肝纤维化的影响.方法将小鼠随机分为A组：手术＋聚肌胞苷酸（Poly I∶C）组;B组：手术组＋磷酸盐缓冲液（PBS）组;C组：假手术＋Poly I∶C组;D组：假手术＋PBS组,每组20只.手术组行胆总管双重结扎术构建梗阻性黄疸模型,假手术组仅游离胆总管.A组、C组每48h腹腔注射1次Poly I∶C,B组、D组注射等量PBS,于第14天处死小鼠,比较各组小鼠血清肝功能指标变化,对肝脏组织行苏木素-伊红（HE）染色、Masson三染色、α-平滑肌肌动蛋白（α-SMA）免疫染色,实时荧光定量聚合酶链反应（FQ-PCR）检测肝脏组织NK细胞表面特异性标志NK1.1、α-SMA及肿瘤坏死因子（TNF）-α mRNA相对表达量.结果 手术组血清肝功能指标高于假手术组,血清谷丙转氨酶（ALT）、谷草转氨酶（AST）水平A组[（829.78±79.33）、（752.00±67.90）U/L]较B组[（508.69±102.46）、（486.20±126.80）U/L]升高（P=0.032,P=0.030）;肝脏组织HE染色、Masson三染色结果表明,A组肝纤维化程度较B组重,α-SMA免疫染色结果根据阳性细胞数及阳性强度评分A组（＋＋＋）、B组（＋＋）、C组（＋）、D组（＋）;NK1.1、α-SMA、肿瘤坏死因子-α（TNF-α） mRNA相对表达量手术组（A组：181.79±13.68、239.67±12.09、12.39±0.87;B组：168.69±12.90、179.96±20.95、7.44±0.66）高于假手术组（C组：91.86±5.95、1.21±0.31、1.51±0.03）,NK1.1 mRNA表达量均较D组升高（P=0.029,P=0.031,P=0.027）.结论 Poly I∶C 诱导肝脏NK细胞活化并肝内聚集,加重了梗阻性黄疸小鼠模型肝纤维化,梗阻性黄疸减弱了肝脏NK细胞的抗肝纤维化的作用.Objective To observe the effect of liver natural killer （NK） cells on a liver fibrosis in mice with obstructive jaundice.Methods Mice were randomly divided into 4 groups with 20 mice in each group： group A, operation ＋ Poly I∶C group;group B, operation ＋ phosphate buffered solution （PBS） group;group C, sham-operation ＋ Poly I∶C group;group D, sham-operation ＋ PBS group.Operation groups underwent common bile duct ligation （BDL） to establish obstructive jaundice models and sham-operation groups only underwent dissociation of common bile duct.Poly I∶C was injected to activate and induce the accumulation of NK cells in mouse liver.All the mice were sacrificed at day 14 after operation.The levels of serum alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, total bilirubin （TBIL）, direct bilirubin （DBIL）, and indirect bilirubin （IBIL） were determined, and the liver tissue received hematoxylin and eosin （HE） stain and Masson stain.The levels of alpha-smooth muscle actin （α-SMA） was detected by histopathologic method.The liver NK 1.1, α-SMA, and tumor necrosis factor-α （TNF-α） mRNA expression was analyzed by real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）.Results As compared with group B [（508.69±102.46）, and （486.20±126.80） U/L] at day 14 after BDL, the serum ALT and AST levels were significantly increased in group A [（829.78±79.33）, and （752.00±67.90） U/L].The results of HE stain and Masson stain showed that the degree of liver fibrosis in the group A was more severe than that in the group B.The liver histopathologic scores of α-SMA in each group were as follows： group A （＋＋＋）, group B （＋＋）, group C （＋）, group D（＋）.As compared with group C （91.86±5.95, 1.21±0.31, and 1.51±0.03） and group D （1.01±0.03, 1.00±0.00, and 0.99±0.02）, the expression levels of NK1.1, α-SMA, and TNF-α mRNA in group A （181.79±13.68, 239.67±12.09, and 12.39±0.87） and gr

关 键 词：梗阻性黄疸 自然杀伤细胞 肝纤维化 

分 类 号：R575[医药卫生—消化系统] R-332[医药卫生—内科学]