摘要
目的:观察C57BL/6N-Tg(1.28HBV)/Vst乙型肝炎病毒(HBV)转基因小鼠复合四氯化碳(CCl 4)腹腔注射诱导乙型肝炎背景下肝纤维化模型小鼠肝内淋巴细胞亚群变化特点,并分析其与血清HBV DNA及肝组织羟脯氨酸(Hyp)含量的相关性。 方法:HBV-Tg小鼠予10% CCl 4腹腔注射诱导加速肝纤维化形成,以血清HBV DNA、HBsAg、HBeAg水平和肝组织HBsAg表达情况评价模型小鼠病毒学特征,肝组织HE、天狼猩红染色及肝组织Hyp含量检测小鼠肝脏炎症及纤维化程度,流式细胞术观察肝内T淋巴细胞、B淋巴细胞、CD4 +T淋巴细胞、CD8 +T淋巴细胞、自然杀伤(NK)细胞、自然杀伤T淋巴(NKT)细胞分布情况。组间数据比较采用单因素方差分析,LSD进行两两比较;Pearson相关性分析上述各淋巴细胞亚群与血清HBV DNA、肝组织Hyp含量的相关性。 结果:HBV-Tg复合CCl 4模型组小鼠血清HBsAg、HBeAg及肝组织HBsAg均呈阳性表达,血清HBV DNA载量> 1×10 6 IU/ml。与野生型对照组相比,复合模型组小鼠肝组织Hyp含量显著升高[(196.39±38.14)μg/g与(347.67±59.53)μg/g, P < 0.01],肝组织炎症及纤维化程度加重,肝内CD4 +T、NK、NKT细胞占淋巴细胞比例均显著减少( P < 0.01),而CD8 +T淋巴细胞(30.58%±2.89%与46.50%±2.24%, P < 0.01)和B淋巴细胞(28.82%±2.24%与37.10%±8.59%, P < 0.05)比例明显增多。血清HBV DNA水平与肝内T淋巴细胞比例呈正相关( r = 0.413, P < 0.05),与NK细胞比例呈负相关( r = -0.419, P < 0.05);肝组织Hyp含量与CD4 +T淋巴细胞( r = -0.871)、NK细胞( r = -0.716)、NKT细胞( r = -0.876)比例均呈负相关( P值均< 0.01),与CD8 +T淋巴细胞( r = 0.852)、B淋巴细胞( r = 0.593)比例均呈正相关( P值均< 0.01)。 结论:HBV-Tg小鼠复合CCl 4造模可诱导HBV病毒学指标阳性、肝脏炎症和纤维化并存的乙型肝炎肝纤维化小鼠模型,该模型具有与人类疾病相似的肝脏淋巴细胞免疫紊乱的特点,且肝内淋巴细胞免疫紊乱与H
Objective To observe the alteration of clinical features of intrahepatic lymphocyte subsets in C57BL/6N-TG(1.28HBV)/Vst hepatitis B virus(HBV)transgenic mice composite carbon tetrachloride(CCl4)with intraperitoneal injection under the background of hepatitis B to induce liver fibrosis mice model,and analyze their correlation with serum HBV DNA and liver tissue hydroxyproline(Hyp)content.Methods HBV-Tg mice were intraperitoneally injected with 10%CCl4 to induce the rapid formation of hepatic fibrosis.Serum HBV DNA,HBsAg,HBeAg levels and liver tissue HBsAg expressional conditions were used to evaluate the virological characteristics of mice model.The degree of hepatic inflammation and fibrosis in mice were observed by HE,Sirius Red staining and liver tissue hydroxyproline(Hyp)content.Intrahepatic T lymphocyte,B lymphocyte,CD4+T lymphocyte,CD8+T lymphocyte,natural killer(NK)cell and natural killer T(NKT)cells distribution were observed by flow cytometry.One-way analysis of variance was used for intergroup data comparison,and LSD was used for pairwise comparison.Pearson’s correlation analysis was used to analyze the correlation between the above lymphocyte subsets and serum HBV DNA and liver tissue Hyp content.Results Serum HBsAg,HBeAg and liver tissue HBsAg had equal positive expression in the HBV-Tg composite CCl4 mice model group,and the serum HBV DNA load was>1×106 IU/ml.Compared with the wild-type control group,liver tissue Hyp content of the composite model group was significantly higher[(196.39±38.14)μg/g and(347.67±59.53)μg/g,P<0.01).The degree of inflammation and fibrosis in liver tissues was aggravated,and the proportion of all intrahepatic CD4+T,NK and NKT cells was significantly reduced(P<0.01),while the proportion of CD8+T lymphocytes(30.58%±2.89%vs.46.50%±2.24%,P<0.01)and B lymphocytes(28.82%±2.24%vs.37.10%±8.59%,P<0.05)was significantly increased.Serum HBV DNA level was positively correlated with the proportion of intrahepatic T lymphocytes(r=0.413,P<0.05),and negatively correlated with the
作者
孙鑫
黄恺
张满
刘洪亮
赵志敏
陶艳艳
彭渊
刘成海
Sun Xin;Huang Kai;Zhang Man;Liu Hongliang;Zhao Zhimin;Tao Yanyan;Peng Yuan;Liu Chenghai(Institute of Liver Diseases,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2020年第7期580-585,共6页
Chinese Journal of Hepatology
基金
国家自然科学基金(81730109)。
关键词
肝炎病毒
乙型
肝纤维化
淋巴细胞
转基因动物
四氯化碳
Hepatitis B virus
Liver fibrosis
Lymphocytes
Transgenic animal
Carbon tetrachloride
