摘要
目的:观察肝凋亡细胞及活性核因子κB在非酒精性脂肪性肝病大鼠模型中的表达及其意义。方法:实验于2005-05/09在解放军南京军区福州总医院动物实验室进行。取40只雄性SD大鼠随机分为2组:造模组30只,给予88%基础饲料+10%猪油+2%胆固醇;正常对照组10只,给予基础饲料。对照组大鼠于8周后处死,造模组分别于8,12,16周处死,取肝组织标本,行苏木精-伊红观察肝组织光镜下的病理改变;苦味酸染色观察肝脏纤维化程度;用TUNEL法检测肝细胞凋亡;活性核因子κB通过免疫组化检测核因子κBp65来表示。采用Pearson相关分析和spearman秩相关分析评估核因子κBp56与肝组织病理学改变及凋亡的相关性。结果:40只大鼠全部进入结果分析。①随着造模时间的延长,造模组大鼠肝组织炎症、脂肪变程度逐渐加重,造模16周窦周出现轻度纤维化(P<0.05)。②造模组12周、16周大鼠肝细胞凋亡细胞数和核因子κBp65阳性细胞数明显多于对照组(P<0.01)。③活化的核因子κB与肝组织炎症、脂肪变及纤维化呈显著正相关(r=0.8491,0.6703,0.4237,P<0.01,0.05);凋亡程度与炎症、脂肪变相关亦呈显著正相关(r=0.6286,0.5936,P<0.01)。④核因子κBp56表达与凋亡存在明显相关关系(r=0.6412,P<0.01)。结论:在非酒精性脂肪性肝病大鼠模型中,肝损伤与肝细胞凋亡的增加明显相关,凋亡与核因子κBp65表达、疾病严重程度相一致。
AIM： To investigate the expression and significance of hepatocyte apoptosis and activation of nuclear＇factor（NF）-gB in rat models with nonalcoholic fatty liver disease.
METHODS： This experiment was conducted at the animal laboratory, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA between May and September 2005, Totally 40 male SD rats were selected and assigned randomly into 2 groups： model group with 30 rats, giving 88% basic feed ＋ 10% pork fat ＋ 2% cholesterol; normal control group with 10 rats, giving basic feed. The rats in the control group were killed 8 weeks later, and those in the model group were killed at the 8^th, 12^th and 16^th weeks to gain hepatic tissue samples. Pathological change of hepatic tissue was observed with hematoxylin-esoin （HE） staining under light microscope. Picric acid staining was applied to observe the degree of hepatic fibrosis. Apoptosis of hepatic cells was detected with TdT-mediated x- dUTP nick end labeling （TUNEL） method. Activated NF-kB was expressed with NF-kBp65 detected by immunohistochemical method. The correlation of NF-kBp65 with pathological change of hepatic tissue and apoptosis was evaluated with Pearson correlation analysis arid spearman rank correlation analysis.
RESULTS： Totally 40 rats entered the analysis of results. ①With the prolonged duration of building models, degrees of inflammation of hepatic tissues and steatosis became aggravated in the model group. At the 16^th week after building models mild fibrosis occurred around sinuses （P 〈 0.05）. ② Numbers of apoptotic cells of hepatic cells and positive cells of NF-kBp65 were significantly more than those in the control group at the 12^th and the 16^th weeks （P 〈 0.01 ）. ③Activated NF-kB had obviously positive correlation with inflammation of hepatic tissues, steatosis and fibrosis （r=0.849 1, 0.670 3,0.423 7,P〈 0.01,0.05）, The degree of apoptosis had markedly positive correlation with inflammation and steatosis （r=0.628 6, 0.593 6,P〈 0.01）. ④The expression of NFkBp56 was dramatically associated with apoptosis （r=0.641 2, P〈 0.01 ）.
CONCLUSION： Liver injury in nonalcoholic fatty liver disease rat models is distinctly associated with the increasing of hepatocyte apoptosis. Further, apoptosis correlates with the expression of NF-kBp65 and disease severity.
出处
《中国临床康复》
CSCD
北大核心
2006年第24期97-99,共3页
Chinese Journal of Clinical Rehabilitation
