胶原纤维在小鼠酒精性肝损伤过程中的表达变化

Changes of collagen fibers in development of alcoholic liver injury

目的:探究胶原纤维在酒精性肝损伤发展过程中各个阶段的变化规律及其与酒精性肝损伤的关系.方法:饲养清洁健康BALB/c♂小鼠40只,随机分为对照组(n=3)与模型组(n=37).将对照组3只小鼠处死,取肝;模型组小鼠每日给予0.15 m L/10 kg 56°红星二锅头白酒灌胃,持续4 wk,于开始灌胃后1、2、3、4 w k各取3只小鼠进行肝脏取材.天狼星红染色法观察小鼠肝组织胶原纤维病理变化情况,蛋白印迹法检测小鼠肝脏增殖细胞核抗原(proliferating cell nuclear antigen,P C N A)蛋白表达及变化趋势.运用ImageProPlus6.0对病理切片样本进行积分光密度(IOD)定量分析,Gel Pro4.0软件对目标蛋白条带灰度值进行测定.运用SPSS16.0对数据进行处理,用ANOVE法和非参数秩和检验法进行显著性分析.结果:第1、2周组小鼠肝组织胶原纤维含量持续显著增高(852.21±10.65 vs 345.24±65.94,1054.15±10.80 vs 852.21±10.65,P<0.01),第2周达到最高,随后第3、4周小鼠肝组织胶原纤维含量依次降低(588.75±17.18 vs 1054.15±10.80,559.40±14.17,P<0.01);小鼠肝组织中PCNA蛋白含量前2 wk明显下降,第2周时达到最小值,第3周较第2周增加显著,差异有统计学意义(P<0.05).第4周与第3周变化无明显差异.结论:在酒精诱导小鼠肝脏损伤的过程中,肝脏胶原纤维的变化规律是先增加后减少,变化明显;PCNA的表达出现显著动态变化,二者均可能与肝脏的损伤修复机制密切相关.

AIM： To observe the change of collagen fibers in the development of alcohol induced liver injury in mice, and investigate the relationship between the change of collage fibers and the stage of alcoholic liver injury. METHODS： Forty BALB/c mice were randomly divided into two groups： a normal group（n = 3） and a model group（n = 37）. Mice of the normal group were sacrificed after adapting the environment for one week, and all the mice of the model group were given 0.15 m L/10 kg Red Star Erguotou（56 degrees） daily by gavage for 4 weeks. Each three mice were sacrificed at 1, 2, 3, and 4 weeks after alcohol administration（n = 37）. Sirius red staining was used to observe collagen fibers in the liver tissue. Western blot was used to detect liver proliferation cell nuclear antigen（PCNA） protein expression. Image-Pro Plus6.0 software was used to analyze the integral optical density（IOD） of the samples, and the gray levels of target protein bands were measured with Gel Pro4.0 software. ANOVE and nonparametric rank sum test were used toanalyze the significance of the data.RESULTS： The contents of collagen fibers increased significantly at 1 and 2 weeks（852.21 ± 10.65 vs 345.24 ± 65.94, 1054.15 ± 10.80 vs 852.21 ± 10.65, P〈0.01）, peaked at 2 weeks, and then decreased at 3 and 4 weeks（588.75 ± 17.18 vs 1054.15 ± 0.80, 559.40 ± 14.17, P〈0.01）. The content of PCNA protein in the liver tissue of mice decreased significantly in the first two weeks, reached the minimum value at 2 weeks, and then increased at 3 and 4 weeks（P〈0.05）.CONCLUSION： In the process of alcohol induced liver injury in mice, the content of collagen fibers in the liver increases first and then decreases, and the expression of PCNA shows significant dynamic changes. The changes may be closely related to the mechanism of liver injury.