肝衰竭前期大鼠模型的建立及辅助性T淋巴细胞17/调节性T淋巴细胞失衡的研究  

Establishment of a rat model of early-stage liver failure and Th17/Treg imbalance

作　　者：李冬[1] 陆忠华[2] 甘建和[1] 

机构地区：[1]苏州大学附属第一医院感染科,江苏苏州215006 [2]江南大学附属无锡市第五人民医院肝病科,江苏无锡214005

出　　处：《临床肝胆病杂志》2016年第5期928-932,共5页Journal of Clinical Hepatology

基　　金：国家"十二五"科技重大专项(2012ZX10002004-008)

摘　　要：目的研究肝衰竭前期大鼠模型建立的方法以及地塞米松和胸腺肽干预前后辅助性T淋巴细胞(Th)17、调节性T淋巴细胞(Treg)和Th17/Treg的变化。方法 64只大鼠随机分为四氯化碳(CCl4)组和脂多糖(LPS)联合D-氨基半乳糖(D-Gal N)组,分别建立肝衰竭前期大鼠模型。通过观察大鼠活动,肝功能以及全血Th17、Treg的变化,评价成模效果。并用地塞米松和胸腺肽进行干预,观察干预效果。计量资料组间比较采用t检验,组内比较采用配对t检验。结果造模组血清ALT、AST、TBil水平显著升高,与对照组相比差异均有统计学意义(CCl4组:t值分别为-3.95、-3.60、-3.57,P值分别为0.006、0.009、0.009;D-Gal N联合LPS组:t值分别为-10.56、-9.63、-11.82,P值均<0.001)。造模后大鼠外周血Th17均上升、Treg均下降,Th17/Treg失衡。造模组大鼠肝组织病理符合肝衰竭前期改变。地塞米松干预后大鼠Th17下降(CCl4组:0.830±0.224 vs 0.580±0.105,t=3.25,P=0.014;D-Gal N联合LPS组:1.301±0.163 vs 0.921±0.141,t=4.12,P=0.004)、Treg上升(CCl4组:0.317±0.076 vs 0.385±0.083,t=-11.13,P<0.001;D-Gal N联合LPS组:0.351±0.110 vs 0.570±0.119,t=-4.06,P=0.005)、Th17/Treg再平衡(CCl4组:2.201±0.556 vs 1.511±0.534,t=3.56,P=0.09;D-Gal N联合LPS组:3.699±0.976 vs 1.619±0.423,t=3.82,P=0.07),胸腺肽干预后Th17上升(CCl4组:1.161±0.219 vs 1.270±0.230,t=-5.74,P=0.001;D-Gal N联合LPS组:0.451±0.095 vs 0.929±0.130,t=-8.61,P<0.001)、Treg下降(CCl4组:0.643±0.100 vs 0.615±0.092,t=2.66,P=0.032;D-Gal N联合LPS组:0.200±0.085 vs 0.161±0.095,t=3.15,P=0.016)、Th17/Treg失衡加剧(CCl4组:1.799±0.625 vs 2.071±0.587,t=-6.47,P<0.001;D-Gal N联合LPS组:2.244±0.634 vs 5.770±1.455,t=-11.72,P<0.001)。结论 CCl4和LPS联合D-Gal N两种方法均可成功建立肝衰竭前期大鼠模型,均未出现死亡,肝组织学及血清生化学变化符合肝衰竭前期改变。地塞米松干预后Th17/Treg失衡改善,胸腺肽干预后动物模型Th17/Treg失衡加�Objective To investigate the methods for establishing a rat model of early- stage liver failure and the changes in Th17,Treg,and Th17 / Treg after dexamethasone and thymosin interventions. Methods A total of 64 rats were randomly divided into carbon tetrachloride（ CCl4） group and endotoxin [lipopolysaccharide（ LPS） ]/D- galactosamine（ D- Gal N） combination group to establish the rat model of early- stage liver failure. The activities of the rats and changes in liver function and whole blood Th17 and Treg were observed to evaluate the effectiveness of the rat model. Dexamethasone and thymosin were used for intervention and related effects were observed. The t- test was used for comparison of continuous data between groups,and the paired t- test was used for comparison of continuous data within one group.Results The serum levels of alanine aminotransferase,aspartate aminotransferase,and total bilirubin showed significant increases in the model groups and were significantly different from those in the control group（ CCl4group： t =- 3. 95,- 3. 60,and- 3. 57,P = 0. 006,0. 009,and 0. 009; LPS / D- Gal N combination group： t =- 10. 56,- 9. 63,and- 11. 82,all P〈0. 001）. After the model was established,the rats showed an increase in Th17,a reduction in Treg,and Th17 / Treg imbalance. The liver pathology in the rats in the model groups was consistent with the changes in early- stage liver failure. After dexamethasone intervention,the rats showed a reduction in Th17（ CCl4group： 0. 830 ± 0. 224 vs 0. 580 ± 0. 105,t = 3. 25,P = 0. 014; LPS / D- Gal N combination group： 1. 301 ± 0. 163 vs 0. 921 ±0. 141,t = 4. 12,P = 0. 004）,an increase in Treg（ CCl4group： 0. 317 ± 0. 076 vs 0. 385 ± 0. 083,t =- 11. 13,P〈0. 001; LPS / D-Gal N combination group： 0. 351 ± 0. 110 vs 0. 570 ± 0. 119,t =- 4. 06,P = 0. 005）,and Th17 / Treg rebalance（ CCl4group： 2. 201 ±0. 556 vs 1. 511 ± 0. 534,t = 3. 56,P = 0. 09; LPS / D- Gal N combination group： 3. 699 ± 0. 976 vs 1. 619 ± 0. 423,t = 3. 82

关 键 词：肝功能衰竭 四氯化碳 脂多糖素类 半乳糖胺 疾病模型 动物 

分 类 号：R575.3[医药卫生—消化系统]