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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
文章发布日期:2016年04月07日  来源:  作者:李冬, 陆忠华,甘建和  点击次数:1243次  下载次数:314次

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【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-GalN)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-GalN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-GalN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-GalN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-GalN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-GalN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-GalN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-GalN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-GalN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失衡加剧。
【Abstract】:ObjectiveTo 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. MethodsA total of 64 rats were randomly divided into carbon tetrachloride (CCl4) group and endotoxin [lipopolysaccharide (LPS)]/D-galactosamine (D-GalN) 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. ResultsThe 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 (CCl4 group: t=-3.95, -3.60, and -3.57, P=0.006, 0009, and 0.009; LPS/D-GalN 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(CCl4 group: 0.830±0.224 vs 0.580±0.105, t=3.25, P=0.014; LPS/D-GalN combination group: 1.301±0.163 vs 0.921±0141, t=4.12, P=0.004), an increase in Treg (CCl4 group: 0.317±0.076 vs 0.385±0.083, t=-11.13, P<0.001; LPS/D-GalN combination group: 0.351±0.110 vs 0.570±0.119, t=-4.06, P=0.005), and Th17/Treg rebalance (CCl4 group: 2.201±0556 vs 1511±0534, t=356, P=009; LPS/D-GalN combination group: 3699±0976 vs 1619±0423, t=382, P=007) After thymosin intervention, the rats showed an increase in Th17 (CCl4 group: 1161±0219 vs 1270±0230, t=-574, P=0001; LPS/D-GalN combination group: 0451±0095 vs 0929±0130, t=-861, P<0001), a reduction in Treg (CCl4 group: 0643±0100 vs 0615±0092, t=266, P=0032; LPS/D-GalN combination group: 0200±0085 vs 0161±0095, t=315, P=0016), and aggravation of Th17/Treg imbalance (CCl4 group: 1799±0625 vs 2071±0587, t=-6.47, P<0.001; LPS/D-GalN combination group: 2.244±0.634 vs 5.770±1.455, t=-11.72, P<0.001). ConclusionThe two methods of CCl4 and LPS/D-GalN combination can successfully establish the rat model of early-stage liver failure with no deaths, and liver histological results and serum biochemical changes are in consistence with the changes in early-stage liver failure. Dexamethasone intervention helps to improve Th17/Treg imbalance, while thymosin intervention causes aggravation of Th17/Treg imbalance.
【关键字】:肝功能衰竭;四氯化碳;脂多糖素类;半乳糖胺;疾病模型,动物
【Key words】:liver failure; carbon tetrachloride; lipopolysaccharide; galactosamine; disease models, animal
【引证本文】:李冬, 陆忠华, 甘建和. 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[J]. 临床肝胆病杂志, 2016, 32(5): 928-932.

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