核自身抗原精子蛋白(NASP)基因突变小鼠伴刀豆球蛋白A(ConA)诱导的肝纤维化加重  

Hepatic fibrosis aggravation in nuclear autoantigenic sperm protein (NASP) mutant mice induced by concanavalin A

作　　者：张艳艳[1] 闫宏伟[2] 朱君 陈龙 王惠 鞠吉雨[1] ZHANG Yanyan;YAN Hongwei;ZHU Jun;CHEN Long;WANG Huil;JU Jiyul(Department of Immunology,Weifang Medical University,Weifang 261053;Department of Blood Transfusion,Affiliated Hospital of Weifang Medical University,Weifang 261031,China)

机构地区：[1]潍坊医学院免疫学教研室,山东潍坊261053 [2]潍坊医学院附属医院输血科,山东潍坊261031

出　　处：《细胞与分子免疫学杂志》2022年第7期577-583,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81373185);山东省自然科学基金(ZR2018MH014)。

摘　　要：目的 探讨核自身抗原精子蛋白(NASP)基因突变对伴刀豆球蛋白A(ConA)诱导的小鼠肝纤维化的影响及其机制。方法 以野生型B6(B6-WT)小鼠为对照组、 NASP基因突变型B6(B6-NASP~M)小鼠为实验组。尾静脉注射ConA每周1次,连续8周,建立肝纤维化模型。HE染色观察小鼠肝组织病理变化,Masson染色观察肝组织胶原纤维沉积;免疫组织化学染色法检测肝组织α平滑肌肌动蛋白(α-SMA)表达;微板法测定血清丙氨酸转氨酶(ALT)和天门冬氨酸转氨酶(AST)水平;ELISA测定血清肿瘤坏死因子α(TNF-α)和γ干扰素(IFN-γ)水平;实时定量PCR检测肝组织Ⅰ型胶原蛋白(Col1)和Col3的mRNA的相对含量;流式细胞术检测肝组织T淋巴细胞亚群变化。结果 与B6-WT组相比,B6-NASP~M组小鼠肝组织结构紊乱、肝组织Col1和Col3的mRNA表达无明显变化,但胶原纤维增生及肝组织α-SMA表达更加明显,血清ALT及TNF-α水平显著升高,肝组织CD4~+CD44T淋巴细胞亚群比例及数量明显下降。结论 NASP基因突变通过增加TNF-α炎症因子释放、改变肝内T淋巴细胞亚群比例以及促进肝星状细胞活化加重小鼠肝纤维化。Objective To investigate how mutation of nuclear autoantigenic sperm protein(NASP) gene affects mouse liver fibrosis induced by concanavalinA(ConA) and its mechanism. Methods The wild-type B6(B6-WT) mice were used as a control group, and the NASP mutant B6(B6-NASP) mice as an experimental group. The mice were injected with ConA via tail vein once a week for 8 weeks to establish the model of liver fibrosis. The histopathological changes were observed by HE staining, the collagen fiber deposition by Masson staining, smooth muscle alpha-actin(α-SMA) expression in liver tissue by immunohistochemical staining. The levels of serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were measured by microplate assay. The serum tumor necrosis factor-alpha(TNF-α) and interferon-gamma(IFN-γ) were measured by ELISA. The mRNA expressions of type I collagen(Col1) and Col3 in liver tissue were determined by real-time quantitative PCR, and the changes of T lymphocyte subsets in liver tissue were detected by flow cytometry. Results Compared with the B6-WT group, B6-NASPgroup had disordered liver structure and no significant changes in mRNA expression of Col 1 and Col3, but the collagen fiber hyperplasia and α-SMA expression in liver tissue were more obvious, and the levels of serum ALT and TNF-α were significantly increased. In addition, the proportion and number of CD4CD44T lymphocyte subsets in liver tissue were markedly decreased. Conclusion Mutation of NASP gene aggravates mouse liver fibrosis by increasing the release of TNF-α, changing the proportion of T lymphocyte subsets in liver tissue and promoting the activation of hepatic stellate cells.

关 键 词：核自身抗原精子蛋白(NASP) 肝纤维化 T淋巴细胞亚群 细胞因子 

分 类 号：R392.12[医药卫生—免疫学] R575[医药卫生—基础医学] R392-33