摘要
目的探讨不同极化方向巨噬细胞对小鼠活化的肝星状细胞(hepatic stellate cell,HSC)凋亡的影响及机制。方法体内构建CCl4诱导的肝纤维化小鼠模型和橄榄油对照组小鼠模型,采用流式细胞术检测不同极化方向巨噬细胞在两组小鼠肝脏中的比例。体外诱导骨髓来源的巨噬细胞极化(M0、M1、M2巨噬细胞),分别收集上清与活化的LX-2细胞共培养。采用CCK-8法、Caspase-3/7活性检测、流式细胞术和Western Blot检测LX-2细胞的凋亡。结果 CCl_4组小鼠肝脏巨噬细胞比例显著高于对照组(t=10.86,P <0.0001),其中M1、M2巨噬细胞的比例均显著高于对照组(t值分别为4.62、5.28,P值分别为0.0036、0.0007)。与M0和M2巨噬细胞共培养组相比,M1巨噬细胞上清可显著抑制LX-2细胞的增殖、诱导LX-2细胞的凋亡、增加Caspase-3/7酶活性并上调凋亡蛋白Bax的表达(P均<0.05)。此外,M1巨噬细胞肿瘤坏死因子相关凋亡诱导配体(TNF-related apoptosis-inducing ligand,TRAIL)的表达水平显著高于M0和M2巨噬细胞(t值分别为17.15、17.20,P均<0.0001)。TRAIL重组蛋白可显著抑制活化的LX-2细胞增殖、促进其凋亡、增强Caspase-3/7酶活性并诱导Bax表达上调。结论 M1巨噬细胞通过表达TRAIL促进小鼠活化的HSC凋亡。
Objective To investigate the effects and its mechanism of different polarization macrophages on the apoptosis of activated hepatic stellate cell(HSC)in mice.Methods Mouse models of CCl4 induced liver fibrosis and olive oil control were constructed in vivo.The proportion of macrophages with different polarization directions in liver of mice in two groups were measured by flow cytometry.In vitro,bone marrow derived macrophages polarization(M0,M1 and M2 macrophage)was induced,and supernatant was collected to co-cultured with activated LX-2 cells.The apoptosis of LX-2 cells was detected by CCK-8,enzyme activity of caspase-3/7,flow cytometry and Western Blot.Results The content of hepatic macrophages of mice in CCl4 group was significantly higher than that in control group(t=10.86,P<0.0001),and the contents of M1 and M2 macrophages were also significantly higher than those in control group(t=4.62,5.28;P=0.0036,0.0007).Compared with the co-culture group of M0 and M2 macrophage,M1 macrophage supernatant significantly inhibited the proliferation of LX-2 cells,induced the apoptosis of LX-2 cells,increased the enzyme activity of Caspase-3/7 and up-regulated the expression of apoptosis protein Bax(P<0.05).In addition,the expression levels of TNF-related apoptosis-inducing ligand(TRAIL)of M1 macrophage were significantly higher than those of M0 and M2 macrophage(t=17.15,17.20;P<0.0001).TRAIL recombinant protein can significantly inhibit the proliferation of activated LX-2 cells,promote their apoptosis,enhance the activity of Caspase-3/7 enzyme and induce the up-regulation of Bax expression.Conclusion M1 macrophage mediate the apoptosis of activated hepatic stellate cells by expressing TRAIL.
作者
麦维利
陈琦琪
杨晓宇
曹颖
朱镠娈
李蕊
闫杰
MAI Wei-Li;CHEN Qi-qi;YANG Xiao-yu;CAO Ying;ZHU Liu-luan;LI Rui;YAN Jie(Department of Liver Diseases ,Peking University Ditan Teaching Hospital,Beijing University,Beijing 100015,China;Department of Liver Diseases ,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Institute of Infectious Diseases/Beijing Key Laboratory of Emerging Sudden Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China)
出处
《中国肝脏病杂志（电子版）》
CAS
2018年第4期59-65,共7页
Chinese Journal of Liver Diseases：Electronic Version
基金
国家自然青年科学基金项目(81600454)
北京市医院管理局"青苗"计划(QML20161802)
