N-乙酰半胱氨酸调控非酒精性脂肪性肝病小鼠外周循环及肝脏局部髓源抑制细胞的研究

doi:10.12092/j.issn.1009-2501.2019.09.005

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (9): 989-996.doi: 10.12092/j.issn.1009-2501.2019.09.005

N-乙酰半胱氨酸调控非酒精性脂肪性肝病小鼠外周循环及肝脏局部髓源抑制细胞的研究

吴加元¹，郑丽¹，莫娟芬¹，郭丽¹，曹晨曦²，鲍轶¹

¹嘉兴市第二医院中心实验室，²肛肠外科，嘉兴 314001，浙江

收稿日期:2019-03-22 修回日期:2019-08-30 出版日期:2019-09-26 发布日期:2019-09-26
通讯作者: 鲍轶，女，主任医师，硕导，研究方向：肿瘤学、免疫学。 Tel: 0573-82082936 E-mail：ybao2011@gmail.com
作者简介:吴加元，男，硕士，药师，研究方向：药理学。 Tel: 0573-82082936 E-mail：jiayuan1100@163.com
基金资助:
浙江省自然科学基金项目（LY16H070007）；浙江省医药卫生科技计划（2016KYB292）；嘉兴市科技计划项目（2018AY32007）

Regulation of N-acetylcysteine on peripheral circulatory and hepatic myeloid-derived suppressor cells in non-alcoholic fatty liver disease mice

WU Jiayuan ¹, ZHENG Li ¹, MO Juanfen ¹, GUO Li ¹, CAO Chenxi², BAO Yi ¹

¹Key Laboratory,² Department of Pathology, the Second Hospital of Jiaxing, Jiaxing 314001, Zhejiang, China

Received:2019-03-22 Revised:2019-08-30 Online:2019-09-26 Published:2019-09-26

摘要/Abstract

摘要：

目的:观察高脂饮食（high fat diet，HFD）诱导的非酒精性脂肪性肝病（non-alcoholic fatty liver disease，NAFLD）小鼠局部及系统性免疫细胞髓源抑制细胞（myeloid-derived suppressor cells，MDSCs）的异常，并探讨N-乙酰半胱氨酸（NAC）对MDSCs及相关促炎细胞因子的调节作用。方法:三组雄性C57B/L6小鼠，每组10只：对照组（Lean）、模型组（HFD）、HFD+NAC给药组（NAC）。16周后，HE与油红O染色观察肝组织病理学改变；流式细胞术结合免疫组化检测小鼠外周血与肝脏中MDSCs的水平；实时荧光定量PCR与蛋白免疫印迹检测肝脏中MDSCs分泌的促炎细胞因子S100钙结合蛋白A8（S100A8）与S100钙结合蛋白A9（S100A9）的改变。结果:与Lean组相比，HFD组小鼠肝发生脂肪变与炎性细胞浸润，外周血CD11b+Gr-1+MDSCs细胞百分比升高（P<0.01），浸润至肝脏的Gr-1+细胞也增加，且肝脏中S100A8与S100A9的mRNA与蛋白水平均升高（P<0.05）。与HFD组对比，NAC改善小鼠肝脂肪变，减少炎性细胞浸润，降低MDSCs细胞百分比（P<0.05），并抑制S100A8与S100A9的表达（P<0.05）。NAC有效抑制肝脏中纤维化相关蛋白α-SMA的水平，缓解肝纤维化。结论:NAC可能通过调控NAFLD小鼠体内MDSCs的异常聚积，抑制炎症反应与纤维化，缓解非酒精性脂肪性肝病的发展进程。

关键词: 髓源抑制细胞, N-乙酰半胱氨酸, 非酒精性脂肪性肝病, 细胞因子

Abstract:

AIM: To observe the local and systemic dysregulation of immune cells myeloid-derived suppressor cells (MDSCs) in high fat diet (HFD) induced non-alcoholic fatty liver disease (NAFLD) mice, and to study the regulation function of N-acetylcysteine (NAC) on MDSCs and associated pro-inflammatory cytokines. METHODS: Thirty male C57B/L6 mice were equally divided into three groups: normal chow diet group (Lean), high fat diet group (HFD) and NAC group (NAC). After 16 weeks, liver pathological change was evaluated by HE and oil red O staining. The levels of MDSCs in peripheral blood and liver tissue were measured through flow cytometry and immunohistochemistry, respectively. The changes of MDSCs secreted pro-inflammatory cytokines S100 calcium-binding protein A8 (S100A8) and S100 calcium-binding proteinA9 (S100A9) in mice liver were tested by real-time quantitative reverse transcription-polymerize chain reaction and western blot. RESULTS:Compared with lean group, HFD group mice exhibited hepatic steatosis and inflammatory cell infiltration. The frequencies of CD11b+Gr-1+ MDSCs in peripheral blood and Gr-1+ cells in liver tissue were elevated in HFD group as well (P<0.01). The mRNA and protein levels of S100A8 and S100A9 also were increased in the livers of HFD group mice (P<0.05). Hepatic steatosis was alleviated and liver-infiltrated inflammatory cells were reduced after NAC treatment. The frequencies of MDSCs in peripheral blood and liver tissue, as well as the levels of S100A8 and S100A9 were decreased by NAC (P<0.05). In addition, NAC effectively suppressed α-SMA level and ameliorated liver fibrosis. CONCLUSION:NAC may slow the progression of NAFLD by regulating MDSCs accumulation, inhibiting inflammation and fibrosis.

Key words: myeloid-derived suppressor cells, N-acetylcysteine, non-alcoholic fatty liver disease, cytokine

中图分类号:

R965.2
R575

吴加元，郑丽，莫娟芬，郭丽，曹晨曦，鲍轶. N-乙酰半胱氨酸调控非酒精性脂肪性肝病小鼠外周循环及肝脏局部髓源抑制细胞的研究[J]. 中国临床药理学与治疗学, 2019, 24(9): 989-996.

WU Jiayuan, ZHENG Li, MO Juanfen, GUO Li, CAO Chenxi, BAO Yi. Regulation of N-acetylcysteine on peripheral circulatory and hepatic myeloid-derived suppressor cells in non-alcoholic fatty liver disease mice[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(9): 989-996.

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N-乙酰半胱氨酸调控非酒精性脂肪性肝病小鼠外周循环及肝脏局部髓源抑制细胞的研究

Regulation of N-acetylcysteine on peripheral circulatory and hepatic myeloid-derived suppressor cells in non-alcoholic fatty liver disease mice

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