重度慢性乙型肝炎患者外周血单个核细胞中焦亡相关因子表达及与预后的关系

doi:10.19871/j.cnki.xfcrbzz.2023.02.006

摘要/Abstract

摘要： 目的探讨重度慢性乙型肝炎（CHB）患者外周血单个核细胞（PBMC）培养上清液中白细胞介素-18（IL-18）、白细胞介素-1β（IL-1β）、Nod样受体蛋白3（NLRP3）、半胱天冬氨酸蛋白酶-1（Caspase-1）表达水平,并分析其与患者预后的关系。方法选取2019年1月至2021年1月海南西部中心医院收治的251例CHB患者作为研究组,根据患者严重程度分为轻度CHB组（93例）、中度CHB组（80例）、重度CHB组（78例）,根据重度CHB患者治疗6个月后是否死亡,将患者分为死亡组（11例）和存活组（67例）,另选取同时期来本院体检的265例健康者作为对照组。采用酶联免疫吸附试验（ELISA）检测PBMC培养上清液中IL-18、IL-1β、NLRP3、Caspase-1表达水平;采用Kaplan-Meier法分析IL-18、IL-1β、NLRP3、Caspase-1水平与重度CHB患者预后的关系;采用多因素Logistic回归分析影响重度CHB患者预后的因素。结果研究组患者PBMC培养上清液中IL-18、IL-1β、NLRP3、Caspase-1水平[（65.86±21.32）pg/ml、（68.06±22.44）pg/ml、（241.57±104.07）pg/ml、（179.01±81.46）pg/ml]均高于对照组[（32.11±8.95）pg/ml、（25.49±7.06）pg/ml、（87.61±23.70）pg/ml、（10.33±3.29）pg/ml],差异有统计学意义（均P<0.05）;轻度CHB组、中度CHB组、重度CHB组患者PBMC培养上清液中IL-18、IL-1β、NLRP3、Caspase-1水平依次升高[IL-18：（50.66±12.80）pg/ml、（70.29±19.03）pg/ml、（79.43±20.56）pg/ml,IL-1β：（51.23±13.17）pg/ml、（72.91±18.64）pg/ml、（83.15±21.78）pg/ml,NLRP3：（136.13±34.95）pg/ml、（289.61±73.08）pg/ml、（318.01±80.50）pg/ml,Caspase-1：（100.26±27.03）pg/ml、（201.30±54.15）pg/ml、（250.04±67.42）pg/ml],差异有统计学意义（均P<0.05）;死亡组患者PBMC培养上清液中IL-18、IL-1β、NLRP3、Caspase-1水平[（104.40±10.36）pg/ml、（110.32±11.57）pg/ml、（367.03±32.25）pg/ml、（292.34±36.28）pg/ml]均高于存活组[（75.33±19.82）pg/ml、（78.69±21.39）pg/ml、（309.96±71.19）pg/ml、（241.10±61.44）pg/ml],差异有统计学意义（均P<0.05）;Kaplan-Meier结果显示,IL-18、IL-1β、NLRP3、Caspase-1高表达的重度CHB患者生存率（75.6%、73.7%、75.6%、76.9%）均低于低表达组（97.3%、97.5%、97.3%、94.9%）（均P<0.05）;终末期肝病模型（MELD）评分、国际标准化比值（INR）及IL-18、IL-1β、NLRP3、Caspase-1水平是影响重度CHB患者预后的独立危险因素（OR=1.821、1.693、2.866、3.514、2.007、2.354,均P<0.05）。结论重度CHB患者PBMC培养上清液中IL-18、IL-1β、NLRP3、Caspase-1水平升高是患者不良预后的独立危险因素,可作为重度CHB患者预后评估的辅助指标。

关键词: 慢性乙型肝炎, 外周血单个核细胞, 焦亡, 预后

Abstract: Objective To investigate the expression levels of interleukin-18 (IL-18), interleukin-1β (IL-1β), nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase (Caspase)-1 in the supernatant of peripheral blood mononuclear cells (PBMC) of patients with severe chronic hepatitis B (CHB), and analyze their relationship with the prognosis of patients. Method A total of 251 CHB patients admitted in Hainan West Central Hospital from January 2019 to January 2021 were gathered as the research group. According to the severity of patients, they were divided into mild CHB group (93 cases), moderate CHB group (80 cases), and severe CHB group (78 cases). According to whether severe CHB patients died after 6 months of treatment, the patients were divided into a death group (11 cases) and a survival group (67 cases). In addition, 265 healthy people who came to our hospital for physical examination during the same period were selected as the control group. Enzyme-linked immunosorbent assay (ELISA method) was used to detect the expression levels of IL-18, IL-1β, NLRP3 and Caspase-1 in the supernatant of PBMC culture; Kaplan-Meier method was used to analyze the relationship between the levels of IL-18, IL-1β, NLRP3, Caspase-1 and the prognosis of patients with severe CHB; multivariate Logistic regression was used to analyze the factors influencing the prognosis of patients with severe CHB. Result The levels of IL-18, IL-1β, NLRP3 and Caspase-1 [(65.86±21.32) pg/ml, (68.06±22.44) pg/ml, (241.57±104.07) pg/ml, (179.01±81.46) pg/ml] in the PBMC culture supernatant of the study group were higher than those in the control group [(32.11±8.95) pg/ml, (25.49±7.06) pg/ml, (87.61±23.70) pg/ml, (10.33±3.29) pg/ml], with statistically significant differences (all P<0.05); the levels of IL-18, IL-1β, NLRP3 and Caspase-1 in the PBMC culture supernatant of patients in mild CHB group, moderate CHB group and severe CHB group increased in turn [IL-18: (50.66±12.80) pg/ml, (70.29±19.03) pg/ml, (79.43±20.56) pg/ml, IL-1β: (51.23±13.17) pg/ml, (72.91±18.64) pg/ml, (83.15±21.78) pg/ml, NLRP3: (136.13±34.95) pg/ml, (289.61±73.08) pg/ml, (318.01±80.50) pg/ml, Caspase-1: (100.26±27.03) pg/ml, (201.30±54.15) pg/ml, (250.04±67.42) pg/ml], with statistically significant differences (all P<0.05); the levels of IL-18, IL-1β, NLRP3 and Caspase-1 [(104.40±10.36) pg/ml, (110.32±11.57) pg/ml, (367.03±32.25) pg/ml, (292.34±36.28) pg/ml] in the PBMC culture supernatant of the death group were significantly higher than those of the survival group [(75.33±19.82) pg/ml, (78.69±21.39) pg/ml, (309.96±71.19) pg/ml, (241.10±61.44) pg/ml], with statistically significant differences (all P<0.05); Kaplan-Meier results showed that the survival rate (75.6%, 73.7%, 75.6%, 76.9%) of patients with severe CHB in the high expression group of IL-18, IL-1β, NLRP3, and Caspase-1 were lower than that in the low expression group (97.3%, 97.5%, 97.3%, 94.9%) (all P<0.05); model for end-stage liver disease (MELD) score, international normalized ratio (INR) and IL-18, IL-1β, NLRP3, Caspase-1 levels were independent risk factors affecting the prognosis of patients with severe CHB (OR=1.821, 1.693, 2.866, 3.514, 2.007, 2.354, all P<0.05). Conclusion The elevated levels of IL-18, IL-1β, NLRP3, and Caspase-1 in the PBMC culture supernatant of patients with severe CHB are related to the poor prognosis of patients, and can be used as auxiliary indicators for prognostic evaluation of patients with severe CHB.

Key words: Chronic hepatitis B, Peripheral blood mononuclear cell, Pyroptosis, Prognosis

中图分类号:

R512.6+2

王莲珊, 马雪梅, 郑成芳. 重度慢性乙型肝炎患者外周血单个核细胞中焦亡相关因子表达及与预后的关系[J]. 新发传染病电子杂志, 2023, 8(2): 28-33.

Wang Lianshan, Ma Xuemei, Zheng Chengfang. Expression of pyroptosis related factors in peripheral blood mononuclear cell of patients with severe chronic hepatitis B and its relationship with prognosis[J]. Electronic Journal of Emerging Infectious Diseases, 2023, 8(2): 28-33.

参考文献

[1] 黄秀玲, 冯程, 董常峰. 超声新技术诊断慢性乙型病毒性肝炎合并非酒精性脂肪性肝病新进展[J/CD]. 新发传染病电子杂志, 2022, 7(2):95-98.
[2] 王亚鹏, 王明良, 侯晓禹, 等. 血常规相关参数在慢性乙型肝炎病毒感染性肝病患者预后中的应用[J]. 临床血液学杂志, 2021, 34(6):432-435,439.
[3] CHEN B, WANG W, XU W, et al.Serum free light chain is associated with histological activity and cirrhosis in patients with chronic hepatitis B[J]. Int Immunopharmaco, 2021, 99(1):107881.
[4] TANG Y, MA T, JIA S, et al.The Mechanism of Interleukin-35 in Chronic Hepatitis B[J]. Semin Liver Dis,2021,41(4):516-524.
[5] SPIEKERMANN C, SEETHALER A, MCNALLY A, et al.Increased levels of S100A8/A9, IL-1ß and IL-18 as a novel biomarker for recurrent tonsillitis[J]. J Inflamm (Lond), 2021, 18(1):24.
[6] LIU Y, MENG X, WANG C, et al.Interleukin-18 Plays a Positive Feedback Activity to Natural Killer-Like B Cells in Hepatitis B Virus-Associated Acute-on-Chronic Liver Failure[J]. Viral Immunol, 2022, 35(1):50-59.
[7] MA T, LIU X, CEN ZF, et al.MicroRNA-302b negatively regulates IL-1β production in response to MSU crystas by targeting IRAK4 and epha2[J]. Arthritis Res Ther, 2018, 20(1):34.
[8] BUSCH K, KNY M, HUANG N, et al.Inhibition of the NLRP3/IL-1β axis protects against sepsis-induced cardiomyopathy[J]. J Cachexia Sarcopenia Muscle, 2021, 12(6):1653-1668.
[9] 石娇, 吴东明, 李静, 等. 痛风性关节炎中IL-1β/Caspase-1介导的细胞焦亡的作用研究[J]. 成都医学院学报,2020, 15(1):19-22.
[10] HUANG Y, XU W, ZHOU R.NLRP3 inflammasome activation and cell death[J]. Cell Mol Immunol, 2021, 18(9):2114-2127.
[11] 中华医学会感染病学分会, 中华医学会肝病学分会. 慢性乙型肝炎防治指南(2019年版)[J/CD]. 中国肝脏病杂志(电子版), 2019, 11(4):5-27.
[12] 中华医学会传染病与寄生虫病学分会, 肝病学分会. 病毒性肝炎防治方案[J]. 中华内科杂志, 2001, 40(1):62-68.
[13] KIM GA, SHIM JJ, LEE JS, et al.Effect of Statin Use on Liver Cancer Mortality Considering Hypercholesterolemia and Obesity in Patients with Non-Cirrhotic Chronic Hepatitis B[J]. Yonsei Med J, 2019, 60(12):1203-1208.
[14] 陈惠珍, 杨筱敏, 卢靖, 等. 重度慢性乙型肝炎预后危险因素及其外周血单个核细胞凋亡分子水平[J]. 中华医院感染学杂志, 2021, 31(18):2734-2737.
[15] XING Z, WU Y, LIU N.IL-22 alleviates the fibrosis of hepatic stellate cells via the inactivation of NLRP3 inflammasomesignaling[J]. ExpTher Med, 2021, 22(4):1088.
[16] 丁霞,廖桂婵,夏牧晔,等.探讨长期核苷(酸)类药物治疗慢性乙型肝炎患者停药后复发的影响因素[J/CD].新发传染病电子杂志,2019,4(1):52-55.
[17] HU Y, YE Y, YE LJ, et al.Efficacy and safety of interferon alpha therapy in children with chronic hepatitis B: a long-term follow-up cohort study from China[J]. Medicine (Baltimore), 2019, 98(32):e16683.
[18] 熊伟,费润欢,周克湘,等.乙型肝炎肝硬化患者血清sICAM-1、IL-6、IL-18及TNF-α水平的表达及临床意义[J]. 现代生物医学进展, 2020, 20(17):3321-3324.
[19] 刘薇拉. 乙型肝炎病毒感染患者外周血白细胞介素-18表达水平与乙型肝炎病毒感染的相关性探讨[J]. 中国药物与临床, 2020, 20(12):1932-1935.
[20] LUAN J, ZHANG X, WANG S, et al.NOD-Like Receptor Protein 3 Inflammasome-Dependent IL-1β Accelerated ConA-Induced Hepatitis[J]. Front Immunol, 2018, 9(1):758.
[21] BEKÇIBASI M, DEVECI Ö, OGUZ A, et al. Serum TNF-α, IL-1β, and IL-6 levels in chronic HBV-infected patients[J]. Int J Clin Pract, 2021, 75(8):e14292.
[22] 王鹏, 郑吉顺, 梁利民. CXCL-13与IL-1β、IL-6在慢性乙型病毒性肝炎患者中表达的临床意义[J]. 河北医药, 2018, 40(13):2018-2020.
[23] 刁凌云, 王胜英, 皇金萍, 等. 慢性乙型肝炎患者血清IL-1β、IL-13与肝硬化的相关性[J]. 现代消化及介入诊疗, 2021, 26(3):352-356.
[24] 吴晓升, 项静, 刘靖华. 线粒体在NLRP3炎性小体激活过程中的双向调节作用[J]. 现代免疫学, 2018, 38(4):333-336.
[25] 麻贞贞, 赵萍, 吕继彩, 等. 系统性红斑狼疮患者外周血单个核细胞中微RNA-223及核苷酸结合寡聚化结构域样受体家族蛋白3炎性小体的表达及临床意义[J]. 中华风湿病学杂志, 2019, 23(1):10-14.
[26] XIE WH, DING J, XIE XX, et al.Hepatitis B virus X protein promotes liver cell pyroptosis under oxidative stress through NLRP3 inflammasomeactivation[J]. Inflamm Res, 2020, 69(7):683-696.
[27] 刘宏伟, 王玉华. 慢性乙型肝炎病人外周血单个核细胞内微小RNA-155、核苷酸结合寡聚化结构域样3表达与乙型肝炎病毒DNA载量的相关性[J]. 安徽医药, 2021, 25(2):373-377.
[28] 龙丹, 韦维. NLRP3/Caspase-1/IL-1β信号通路在肝纤维化中的作用研究进展[J]. 中华中医药学刊, 2022, 40(1):75-79.
[29] 窦增花, 于国英. 沙棘熊果酸调控NLRP3/caspase-1轴在抗结核药物致人肝细胞损伤中的作用[J]. 中成药, 2021, 43(5):1319-1323.