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目的研究小鼠酒精性肝损伤过程中肝细胞凋亡的变化规律。方法按照体重将健康KM雄性小鼠随机分为2组:对照组(n=10),模型组(n=80)。模型组给予56°白酒(7.5μL·g~(-1)·d~(-1))灌胃,持续8周。分别于1,2,3,4,5,6,7,8周脱臼处死小鼠,取肝组织;对照组小鼠等量蒸馏水灌胃后处死。进行苏木素-伊红染色(HE)和Hoechst_33258染色,检测小鼠酒精性肝损伤中肝细胞的凋亡变化;荧光显微镜下辨认和计数正常肝细胞和凋亡肝细胞,计算细胞凋亡率。结果第3周,模型组小鼠发生急性肝损伤改变,表现为炎症细胞浸润加剧;到第8周,进展为慢性肝损伤,表现为肝组织水样变,脂肪空泡。第1,2,3周,模型组的凋亡率分别为(9.28±0.41)%,(13.20±0.81)%,(20.43±0.97)%,与对照组的(1.38±0.18)%比较,差异均有统计学意义(均P<0.05)。第8周末,凋亡率达到(51.99±1.70)%,与对照组相比,差异有统计学意义(P<0.01)。结论小鼠酒精性肝损伤程度与肝细胞凋亡的发生发展密切相关。
Objective To study the changes of hepatocyte apoptosis during alcoholic liver injury in mice. Methods Healthy KM male mice were randomly divided into 2 groups according to body weight: control group (n = 10) and model group (n = 80). The model group was intragastrically given 56 ° liquor (7.5μL · g -1 d -1) for 8 weeks. Mice were sacrificed at 1, 2, 3, 4, 5, 6, 7 and 8 weeks after dislocation, and the liver tissues were removed. The control mice were sacrificed by equal dosage of distilled water. The hematoxylin-eosin staining (HE) and Hoechst_33258 staining were used to detect the changes of hepatocytes in mice with alcoholic liver injury. The normal hepatocytes and apoptotic hepatocytes were identified and counted under fluorescence microscope, and the apoptosis rate was calculated. Results At the third week, the mice in the model group changed acute liver injury, showing the aggravation of the infiltration of inflammatory cells. By the eighth week, the mice developed chronic liver injury, manifested as watery changes of the liver tissue and vacuoles. The apoptotic rates in the model group were (9.28 ± 0.41)%, (13.20 ± 0.81)% and (20.43 ± 0.97)%, respectively, compared with those in the control group (1.38 ± 0.18)% All were statistically significant (P <0.05). At the end of the 8th week, the apoptotic rate reached (51.99 ± 1.70)%, which was significantly different from the control group (P <0.01). Conclusion The degree of alcoholic liver injury in mice is closely related to the occurrence and development of hepatocyte apoptosis.