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肝泡型包虫病患者中性粒细胞胞外诱捕网的表达及吞噬功能

[Expression of neutrophil extracellular traps and phagocytic functions among patients with hepatic alveolar echinococcosis].

作者信息

Renzeng Z, Fan H, Yang K, Wang Z, Zhang Y, Lu Y, Wang H

机构信息

Department of Hepatobiliary Surgery, The Affiliated Hospital of Qinghai University, Xining, Qinghai 810001, China.

Qinghai Provincial Key Laboratory of Hydatid Disease Research, Xining, Qinghai 810001, China.

出版信息

Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2024 Feb 26;36(1):25-33. doi: 10.16250/j.32.1374.2023172.

Abstract

OBJECTIVE

To investigate the expression of neutrophil extracellular traps (NETs) and phagocytic function in the peripheral blood of patients with hepatic alveolar echinococcosis (HAE), and to examine their correlations with clinical inflamma tory indicators and liver functions.

METHODS

A total of 50 patients with HAE admitted to Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qinghai University from August 2022 to June 2023 were enrolled, while 50 age- and gender-matched healthy individuals from the Centre for Healthy Examinations of the hospital during the same period served as controls. The levels of NETs markers neutrophil myeloperoxidase (MPO) and neutrophil elastase (NE) were measured using enzyme-linked immunosorbent assay (ELISA). Peripheral blood neutrophils were isolated using density gradient centrifugation, stimulated using phorbol 12-myristate 13 acetate (PMA), and the levels of MPO and citrullination histone H3 (CitH3) released by neutrophils were quantified using flow cytometry. The phagocytic functions of neutrophils were examined using flow cytometry. In addition, the correlations of MPO and NE levels with clinical inflammatory indicators and liver biochemical indicators were examined using Spearman correlation analysis among HAE patients.

RESULTS

The peripheral blood plasma MPO[(417.15 ± 76.08) ng/mL vs. (255.70 ± 80.84) ng/mL; = 10.28, < 0.05], NE[(23.16 ± 6.75) ng/mL vs. (11.92 ± 3.17) ng/mL; = 10.65, < 0.05]and CitH3 levels[(33.93 ± 18.93) ng/mL vs. (19.52 ± 13.89) ng/mL; = 4.34, < 0.05]were all significantly higher among HAE patients than among healthy controls, and a lower phagocytosis rate of neutrophils was detected among HAE patients than among healthy controls[(70.85 ± 7.32)% vs. (94.04 ± 3.90)%; = 20.18, < 0.05], and the ability to produce NETs by neutrophils was higher among HAE patients than among healthy controls following PMA stimulation. Pearson correlation analysis showed that the phagocytosis rate of neutrophils correlated negatively with platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), interleukin-6 (IL-6) level and C-reactive protein (CRP) level ( = -0.515 to -0.392, all values < 0.05), and the MPO and NE levels positively correlated with inflammatory markers NLR, PLR, CRP and IL-6 ( = 0.333 to 0.445, all values < 0.05) and clinical liver biochemical indicators aspartic transaminase, alanine aminotransferase, direct bilirubin and total bilirubin among HAE patients ( = 0.290 to 0.628, all values < 0.001).

CONCLUSIONS

Excessive formation of NETs is found among HAE patients, which affects the phagocytic ability of neutrophils and results in elevated levels of inflammatory indicators. NETs markers may be promising novel biomarkers for early diagnosis, monitoring, and severity assessment of liver disease.

摘要

目的

探讨肝泡型包虫病(HAE)患者外周血中性粒细胞胞外陷阱(NETs)的表达及吞噬功能,并检测其与临床炎症指标及肝功能的相关性。

方法

选取2022年8月至2023年6月青海大学附属医院肝胆胰外科收治的50例HAE患者,同期选取该医院健康体检中心50例年龄、性别匹配的健康个体作为对照。采用酶联免疫吸附测定(ELISA)法检测NETs标志物中性粒细胞髓过氧化物酶(MPO)和中性粒细胞弹性蛋白酶(NE)水平。采用密度梯度离心法分离外周血中性粒细胞,用佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯(PMA)刺激,采用流式细胞术定量检测中性粒细胞释放的MPO和瓜氨酸化组蛋白H3(CitH3)水平。采用流式细胞术检测中性粒细胞的吞噬功能。此外,在HAE患者中采用Spearman相关分析检测MPO和NE水平与临床炎症指标及肝脏生化指标的相关性。

结果

HAE患者外周血血浆MPO[(417.15±76.08)ng/mL对(255.70±80.84)ng/mL;t = 10.28,P < 0.05]、NE[(23.16±6.75)ng/mL对(11.92±3.17)ng/mL;t = 10.65,P < 0.05]和CitH3水平[(33.93±18.93)ng/mL对(19.52±13.89)ng/mL;t = 4.34,P < 0.05]均显著高于健康对照,且HAE患者中性粒细胞吞噬率低于健康对照[(70.85±7.32)%对(94.04±3.90)%;t = 20.18,P < 0.05],PMA刺激后HAE患者中性粒细胞产生NETs的能力高于健康对照。Pearson相关分析显示,中性粒细胞吞噬率与血小板/淋巴细胞比值(PLR)、中性粒细胞/淋巴细胞比值(NLR)、白细胞介素 - 6(IL - 6)水平及C反应蛋白(CRP)水平呈负相关(r = -0.515至 -0.392,均P值 < 0.05),在HAE患者中MPO和NE水平与炎症标志物NLR、PLR、CRP和IL - 6呈正相关(r = 0.333至0.445,均P值 < 0.05),与临床肝脏生化指标天冬氨酸转氨酶、丙氨酸转氨酶、直接胆红素和总胆红素也呈正相关(r = 0.290至0.628,均P值 < 0.001)。

结论

HAE患者存在NETs过度形成,影响中性粒细胞吞噬能力,导致炎症指标升高。NETs标志物可能是有前景的用于肝脏疾病早期诊断、监测及严重程度评估的新型生物标志物。

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