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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.
Objective: To investigate the changes of collagen fibers in various stages of development of alcoholic liver injury and its relationship with alcoholic liver injury.Methods: Forty healthy and healthy BALB / c♂ mice were housed and randomly divided into control group (n = 3) and model group (n = 37). Three mice in the control group were sacrificed and the liver was taken. The model mice were orally administered with 0.15 m L / 10 kg 56 ° red star Erguotou liquor daily for 4 weeks, Three mice were harvested at 1, 2, 3 and 4 wk after gavage, respectively.The pathological changes of collagen fibers in liver were observed by Sirius red staining, and the proliferating cell nuclear antigen, PCNA) protein expression and trend of changes.Using ImageProPlus6.0 for pathological sections of the quantitative optical density (IOD) quantitative analysis, Gel Pro4.0 software for the determination of the target protein band gray value.Using SPSS16.0 pairs of data And the ANOVA and nonparametric rank sum test were used for significance analysis.Results: The content of collagen fiber in liver tissue of mice in the first and second week continued to increase significantly (852.21 ± 10.65 vs 345.24 ± 65.94, 1054.15 ± 10.80 vs 852.21 ± 10.65, P <0.01), reached at the second week The content of collagen fibers in the liver tissues of mice in the first and third week were decreased (588.75 ± 17.18 vs 1054.15 ± 10.80, 559.40 ± 14.17, P <0.01) (P <0.05), there was no significant difference between the 4th week and the 3rd week.Conclusion: In the alcohol-induced liver injury in mice During the process, the changes of hepatic collagen fibers firstly increased, then decreased and changed obviously. The expression of PCNA showed significant dynamic changes, both of which may be closely related to the liver injury and repair mechanism.