CXC趋化因子配体12及其CXC趋化受体4/7在肝硬化脾亢大鼠脾脏组织中的表达及意义  

The expression of chemokine CXC motif ligand 12, CXC chemokine receptor 4 and CXC chemokine receptor 7 in spleens of rats with cirrhosis and hypersplenism

作　　者：黎业娟[1,2] 吕云福[2] 韩晓玉[2] 刘宁[2] 邓杰[1,2] 李晴晴[1,2] 

机构地区：[1]海南大学,海口570228 [2]海南省人民医院普外科,海口570311

出　　处：《中华实验外科杂志》2017年第7期1122-1126,共5页Chinese Journal of Experimental Surgery

基　　金：海南省科技合作专项资金项目（KJHZ2015-28）;海南省重点科技研发项目（S92016SHFZ0044）

摘　　要：目的探讨CXC趋化因子配体12（CXCL12）及其CXC趋化受体4/7（CXCR4/7）在肝硬化脾亢大鼠模型巨脾组织中的表达及意义。方法将50只雄性SD纯系大鼠，随机分成模型组（n=40）和对照组（n=10）。对照组采用0.9%生理盐水灌胃，剂量为0.3 ml/100 g，每周2次，共8周；模型组则采用40%四氯化碳（CCL4）花生油溶液灌胃，剂量0.3 ml/100 g，每周2次，共8周。经病理学和血常规检查证实制成肝硬化脾亢模型后，用Masson三色染色观察脾脏组织纤维化程度，用免疫组织化学，Western blot和实时定量反转录聚合酶链反应（RT-qPCR）三方法检测脾脏组织中趋化轴CXCL12-CXCR4/CXCR7的表达水平，并与对照组进行比较。结果Masson三色染色显示：肝硬化脾亢大鼠脾脏纤维面积[（11.17±4.85）%]显著高于对照组[（2.83±0.90）%]，两组比较差异有统计学意义（t=7.939，P=0.000）。模型组脾脏指数[（3.14±0.98） mg/g]高于对照组[（2.33±1.03） mg/g]，两者比较差异有统计学意义（t=2.351，P=0.025）。免疫组织化学检测：模型组CXCL12、CXCR4及CXCR7的平均吸光度值分别为（8.89±4.89）×10^－3、（8.78±4.67）×10^－3、（8.47±3.97）×10^－3，显著高于对照组（2.89±1.11）×10^－3、（2.03±0.45）×10^－3、（1.81±0.69）×10^－3，差异有统计学意义（t=4.126，P=0.001；t=4.973，P=0.000；t=5.644，P=0.000）；模型组CXCL12、CXCR4及CXCR7阳性细胞率分别为（31.02±9.03）%、（33.22±8.31）%、（28.89±8.40）%，显著高于对照组（20.62±8.88）%、（18.81±6.23）%、（10.49±2.99）%，差异有统计学意义（t=3.112，P=0.004；t=4.170，P=0.001；t=6.293，P=0.000）。Western blot检测：模型组CXCL12、CXCR4及CXCR7蛋白相对表达量分别为1.71±0.98、1.01±0.57、1.15±0.77，显著高于对照组0.39±0.13、0.32±0.11、0.51±0.31，差异有统计学意义（t=6.366，P=0.000；t=5.550，P=0.000；t=3.348，P=0.002）。RT-qPCR检�Objective To investigate expression the of CXC motif ligand 12 （CXCL12）-CXC chemokine receptor 4 （CXCR4）/CXC chemokine receptor 7 （CXCR7） axis in the spleens of rats with cirrhosis and hypersplenism, provide theoretical basis for the further explore the mechanism of spleen fibrosis of cirrhosis and hypersplenism.Methods Fifty male SD rats were randomized into 2 groups： In normal control group （n=10） were fed normal saline （0.3 ml/100 g （twice per week for 8 weeks）; In model group （n=40） were fed with the volume fraction of 40% carbon tetrachloride （CCL4） of peanut oil solution （0.3 ml/100 g, twice per week for 8 weeks）. The establishment of animal models with cirrhotic hypersplenic models were indicated by pathology and hemogram.. Spleen fibrosis degree was assessed by Masson trichrome staining. The expression of chemokine receptors of CXCL12, CXCR4 and CXCR7 were investigated by immunohistochemical staining, Western blotting analysis and real-time quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR）. The comparison between the two groups were used the independent sample T test.Results Masson trichrome staining displayed that collagen fiber area in the spleen of rats induced by CCL4 is remarkably larger than the controls [（11.17±4.85）% vs. （2.83±0.90）%, t=7.939, P=0.000]. There was significance differences （t=2.351, P=0.025） in the spleen index between the models （3.14±0.98） mg/g and the controls （2.33±1.03） mg/g. Immunohistochemical staining revealed that the mean density of CXCL12, CXCR4 and CXCR7 was significantly increased in models compared with the controls [（8.89±4.89）×10^-3s. （2.89±1.11）×10^-3,t=4.126, P=0.001; （8.78±4.67）×10^-3vs. （2.03±0.45）×10^-3,t=4.973, P=0.000; （8.47±3.97）×10^-3 vs. （1.81±0.69）×10^-3,t=5.644, P=0.000; respectively]. Meanwhile, the models displayed a higher rate of positive cells of CXCL12, CXCR4 and CXCR7 [（31.02±9.03）%vs. （20.62±8.88）%, t=3.112, P=0.004; �

关 键 词：趋化轴CXC趋化因子配体12-CXC趋化受体4/CXC趋化受体7 脾脏纤维化 肝硬化 脾亢 模型 动物 人鼠 

分 类 号：R575.2[医药卫生—消化系统]