Page 33 - 南京医科大学学报自然科学版

第41卷第5期 南京医科大学学报（自然科学版）
2021年5月 Journal of Nanjing Medical University（Natural Sciences） ·663 ·

·基础医学·

NEK7在慢性肝脏炎症及纤维化形成中的作用

王艺睿，王 皓，刘硕琛，倪 鸣，李相成 *
南京医科大学第一附属医院肝胆中心，江苏 南京 210029

［摘 要］ 目的：探讨肝脏巨噬细胞中NIMA相关蛋白激酶7（never in mitosis gene a⁃related kinase 7，NEK7）在肝脏纤维化组织
中的表达及其通过对NOD样受体家族3（nucleotide⁃binding oligomerization domain⁃like receptors，NLRP3）炎症小体信号通路调
控肝脏纤维化（liver fibrosis）的作用机制。方法：通过HE和Masson染色以及qRT⁃PCR技术观察对比正常人和肝纤维化患者之
间肝脏纤维化程度的差异；采用Western blot和qRT⁃PCR技术检测正常人和肝纤维化患者肝脏组织中NEK7的表达；运用免疫
荧光染色技术观察CD68阳性细胞中NEK7的表达情况。通过腹腔注射四氯化碳（carbon tetrachloride，CCl4 ）8周的方法建立小
鼠肝脏纤维化模型，采用巨噬细胞特异性干扰RNA（small interfering RNA，siRNA）敲低巨噬细胞中NEK7的表达，取组织标本，
通过HE、Masson和天狼星红染色技术以及qRT⁃PCR技术，观察小鼠肝脏组织纤维化程度的差异；同时利用组织免疫化学染色
和组织免疫荧光染色技术，观察小鼠肝脏炎症反应中中性粒细胞和巨噬细胞聚集程度的差异；进一步运用Western blot技术检
测其对NLRP3炎症小体信号通路的影响。结果：NEK7在肝纤维化肝脏组织中表达明显高于正常肝脏组织；在体内特异性敲
低巨噬细胞中NEK7的表达有助于减轻肝脏纤维化的程度和炎症反应以及抑制NLRP3炎症小体信号通路的激活。结论：抑制
巨噬细胞中NEK7的表达能通过抑制NLRP3炎症小体信号通路，减轻炎症反应，阻止肝纤维化病变。
［关键词］ NEK7；纤维化；NLRP3炎症小体信号通路；肝硬化

［中图分类号］ R575.2 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）05⁃663⁃08
doi：10.7655/NYDXBNS20210505

NEK7 in chronic liver inflammatory and liver fibrosis
WANG Yirui，WANG Hao，LIU Shuochen，NI Ming，LI Xiangcheng *
Hepatobiliary Center，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，China

［Abstract］ Objective：This study aims to investigate the never in mitosis gene a⁃related kinase 7（NEK7）expression in liver fibrosis
and its effect on nucleotide⁃binding oligomerization domain⁃like receptors（NLRP3）signal pathway to regulate liver fibrosis. Methods：
HE staining，Masson staining and quantitative reverse transcription PCR（qRT⁃PCR）were used to evaluate those degree of hepatic
fibrosis in normals and patients with cirrhosis. The expression of NEK7 in liver tissues of non⁃cirrhotic and cirrhotic patients was
detected by Western blot and qRT⁃PCR. CD68⁃positive cells was determined by immunofluorescence tissue staining. To induce hepatic
fibrosis，C57BL/6J mice were injected intraperitoneally with carbon tetrachloride（CCL4 ）twice a week for 8 weeks. C57BL/6J mice were
injected through tail vein with macrophage⁃specific siRNA to knockdown the expression of NEK7 in macrophages. HE staining，Masson
staining and sirius red staining were used to observe the changes of liver fibrosis in mice liver tissues. The level of the migration and
aggregation of neutrophils and macrophages were monitored by immunohistochemistry tissue staining and immunofluorescence tissue
staining. The impacts of NEK7 on NLRP3 signaling pathway were measured by Western blot. Results：In the fibrotic liver tissues，
NEK7 expression was significantly higher than that in the normal tissues. Specific blockade of NEK7 in macrophages obviously
attenuated liver fibrosis and hepatic inflammation in vivo. Specific blockade of NEK7 in macrophages also inhibited the activation of
NLRP3 inflammasome signaling pathway. Conclusion：Inhibition of NEK7 may alleviate macrophage inflammation reaction and liver
fibrosis through inhibiting NLRP3 inflammasome signaling pathway.
［Key words］ NEK7；liver fibrosis；NLRP3 inflammasome signaling pathway；liver cirrhosis
［J Nanjing Med Univ，2021，41（05）：663⁃669，689］

［基金项目］ 国家自然科学基金项目（81670570）；江苏省重点病种规范化诊疗研究项目（BE2016789）
∗
通信作者（Corresponding author），E⁃mail： drxcli@njmu.edu.cn