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腹透液来源外泌体中的糖蛋白 96：评估腹膜转运特性和炎症状态的工具。

Glycoprotein 96 in Peritoneal Dialysis Effluent-Derived Extracellular Vesicles: A Tool for Evaluating Peritoneal Transport Properties and Inflammatory Status.

机构信息

Division of Nephrology and Unit of Critical Nephrology, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

Research and Development Center, Shanghai Applied Protein Technology Co., Ltd., Shanghai, China.

出版信息

Front Immunol. 2022 Feb 10;13:824278. doi: 10.3389/fimmu.2022.824278. eCollection 2022.

DOI:10.3389/fimmu.2022.824278

PMID:35222405

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8866190/

Abstract

BACKGROUND

Extracellular vesicles (EVs) from peritoneal dialysis effluent (PDE), containing molecules such as proteins and microRNAs (miRNAs), may be potential biological markers to monitor peritoneal function or injury. Peritoneal inflammation is an important determinant of peritoneal solute transport rate (PSTR). Thus, the aim of this study is to determine whether the specific proteins capable of evaluating the PSTR could be found in PDE-EVs, and explore the underlying mechanism for the association between PSTR and peritoneal inflammation.

METHODS

Sixty patients undergoing peritoneal dialysis (PD) were divided into two groups: high/high average transport (H/A) group (PET >0.65) and low/low average transport (L/A) group (PET <0.65). EVs derived from PDE (PDE-EVs) were isolated by ultracentrifugation. Proteomic analysis was performed to explore the differentially expressed proteins and identify the potential biomarkers in PDE-EVs from the two groups, and we focused on glycoprotein 96 (GP96) as it could be involved in the inflammatory process. The expression of GP96 in PDE-EVs and inflammatory cytokines was quantified by real-time PCR and enzyme-linked immunosorbent assay. The infiltration of macrophages and neutrophils into the peritoneum was detected using immunohistochemistry in a PD rat model.

RESULTS

The expression of PDE-EVs-GP96 was significantly higher in the H/A group, and was positively correlated with the PSTR and the level of the inflammatory factor interleukin (IL)-6. GP96-enriched EVs enhanced the secretion of proinflammatory cytokines IL-1β, IL-6, tumor necrosis factor (TNF)-α, and IL-8 in macrophages, which was reversed by a pharmacological GP96-specific inhibitor (PU-WS13). The GP96 inhibitor also reduced local peritoneal inflammation by decreasing the infiltration of inflammatory cells and levels of proinflammatory cytokines (IL-6 and TNF-α) and chemokines (CCL2, CXCL1, and CXCL2) in a PD rat model.

CONCLUSIONS

PDE-EVs-GP96 is a new promising tool to evaluate the status of peritoneal inflammation and PSTR, and the mechanism may be related to affecting the inflammatory properties of macrophages.

摘要

背景

腹腔透析液（PDE）中的细胞外囊泡（EVs）含有蛋白质和 microRNAs（miRNAs）等分子，可能是监测腹膜功能或损伤的潜在生物标志物。腹膜炎症是腹膜溶质转运率（PSTR）的重要决定因素。因此，本研究旨在确定 PDE-EVs 中是否存在能够评估 PSTR 的特定蛋白质，并探讨 PSTR 与腹膜炎症之间关联的潜在机制。

方法

将 60 名接受腹膜透析（PD）的患者分为两组：高/高平均转运（H/A）组（PET>0.65）和低/低平均转运（L/A）组（PET<0.65）。通过超速离心法从 PDE 中分离 EVs（PDE-EVs）。进行蛋白质组学分析以探讨两组 PDE-EVs 中差异表达的蛋白质，并鉴定潜在的生物标志物，我们重点关注糖蛋白 96（GP96），因为它可能参与炎症过程。通过实时 PCR 和酶联免疫吸附试验定量测定 PDE-EVs 中 GP96 和炎症细胞因子的表达。通过 PD 大鼠模型的免疫组织化学检测巨噬细胞和中性粒细胞浸润到腹膜中。

结果

H/A 组中 PDE-EVs-GP96 的表达明显升高，并且与 PSTR 和炎症因子白细胞介素（IL）-6 的水平呈正相关。富含 GP96 的 EVs 增强了巨噬细胞中促炎细胞因子 IL-1β、IL-6、肿瘤坏死因子（TNF）-α 和 IL-8 的分泌，而 GP96 特异性抑制剂（PU-WS13）可逆转这一作用。GP96 抑制剂还通过减少炎症细胞浸润和降低促炎细胞因子（IL-6 和 TNF-α）和趋化因子（CCL2、CXCL1 和 CXCL2）的水平，减少 PD 大鼠模型中的局部腹膜炎症。