Department of Molecular and cellular medicine, Institute of Liver and Biliary Sciences, Delhi, India.
Department of Pathology, Institute of Liver and Biliary Sciences, Delhi, India.
Aliment Pharmacol Ther. 2018 Apr;47(8):1151-1161. doi: 10.1111/apt.14564. Epub 2018 Feb 20.
Severe alcoholic hepatitis patients have high mortality and limited response to corticosteroids. Microvesicles reflect cellular stress and disease conditions.
To investigate whether microvesicles are associated with severity, response to steroid therapy and inflammation in severe alcoholic hepatitis.
Microvesicles originating from different cells were studied pre-therapy in 101 patients; (71 responder to corticosteroid therapy and 30 nonresponders) and 20 healthy controls. Microvesicles and cells were determined in peripheral and hepatic vein samples using flow cytometry and correlated with outcomes. Inflammatory signalling pathways and functional alterations of immune cells after stimulation with microvesicles were also investigated.
Microvesicles mean levels were higher in nonresponders for T cells (CD3 CD4 ; 10.1 MV/μL vs 5.4; P = 0.06), macrophages (CD68 CD11b ; 136.5 vs 121.9 MV/μL; P = 0.01), haematopoietic stem-cells (CD45 CD34 ; 116.8 vs 13.4 MV/μL; P = 0.0001) and hepatocytes (ASGPR ; 470 vs 361 MV/μL; P = 0.01); the latter two predicting steroid nonresponse in 94% patients at baseline in peripheral plasma. Microvesicle levels correlated with histological and liver disease severity indices. Whereas, in non-responders hepatic vein CD34+ cells were lower (P = 0.02), the CD34+ microvesicles there from were higher (P = 0.04), thus suggesting impaired regeneration. Also, microvesicles of 0.2-0.4 μm size were higher in nonresponders (P < 0.03) at baseline. Microvesicles from patients trigger more (P = 0.04) ROS generation, TNF-α production (P = 0.04) and up-regulate pro-inflammatory cytokine related genes in neutrophils in vitro.
Pre-therapy peripheral plasma levels of CD34 and ASGPR microvesicles are reliable non-invasive markers of steroid nonresponse and mortality in patients with severe alcoholic hepatitis.
重症酒精性肝炎患者死亡率高，对皮质类固醇的反应有限。微泡反映细胞应激和疾病状况。
研究微泡是否与重症酒精性肝炎的严重程度、对类固醇治疗的反应和炎症有关。
在 101 例患者（71 例对皮质类固醇治疗有反应，30 例无反应）和 20 例健康对照者中，治疗前检测了来自不同细胞的微泡。使用流式细胞术在周围静脉和肝静脉样本中检测微泡和细胞，并与结果相关联。还研究了微泡刺激后免疫细胞的炎症信号通路和功能改变。
无反应者的 T 细胞（CD3+CD4+）、巨噬细胞（CD68+CD11b+）、造血干细胞（CD45+CD34+）和肝细胞（ASGPR+）的微泡平均水平较高（分别为 10.1MV/μL、136.5MV/μL、116.8MV/μL 和 470MV/μL）（P<0.06、P=0.01、P=0.0001 和 P=0.01）；这两种细胞在基线时可预测 94%的外周血患者对皮质类固醇的无反应。微泡水平与组织学和肝病严重程度指数相关。然而，无反应者肝静脉 CD34+细胞较低（P=0.02），但那里的 CD34+微泡较高（P=0.04），提示再生受损。此外，无反应者的 0.2-0.4μm 大小的微泡基线较高（P<0.03）。患者的微泡引发更多的 ROS 生成（P=0.04）、TNF-α 产生（P=0.04），并在体外上调中性粒细胞中与促炎细胞因子相关的基因。
重症酒精性肝炎患者治疗前外周血 CD34 和 ASGPR 微泡水平是预测皮质类固醇无反应和死亡率的可靠非侵入性标志物。
