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不同阶段慢性乙型肝炎患者外周血单个核细胞中NF-κB/iNOS的表达及免疫状态
Expression of nuclear factor-kappa B and inducible nitric oxide synthase in peripheral blood mononucleated cells and immune status in patients with different stages of chronic hepatitis B
文章发布日期:2020年02月17日  来源:  作者:黄津,张美泉,何道兴  点击次数:291次  下载次数:72次

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【摘要】:目的 研究核因子-κB(NF-κB)/诱导型一氧化氮合成酶(iNOS)在不同阶段慢性乙型肝炎患者外周血单个核细胞(PBMC)中的表达,比较各阶段自然杀伤细胞及T、B淋巴细胞的变化情况。方法 纳入福建医科大学附属南平第一医院2017年10月-2019年1月慢性乙型肝炎相关疾病患者,按照疾病不同阶段分4组:肝炎组、肝衰竭组、肝硬化组、肝癌组,每组各20例;另选取10例健康志愿者作为对照组。采用流式细胞仪检测5组外周血总淋巴细胞计数和淋巴细胞亚群计数;采用Western Blot检测PBMC中NF-κB、iNOS蛋白的表达水平;硝酸还原酶法和ELISA法检测血清中NO、IL-6水平。多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 与对照组比较,肝炎组、肝衰竭组、肝癌组、肝硬化组总淋巴细胞计数水平均明显下降(P值均<0.05);与肝炎组比较,肝衰竭组、肝癌组、肝硬化组总淋巴细胞计数水平均明显下降(P值均<0.05)。肝衰竭组、肝癌组、肝硬化组CD3+明显下降,分别与对照组、肝炎组比较差异均有统计学意义(P值均<0.05);肝衰竭组、肝癌组、肝硬化组与肝炎组CD4+比较明显下降(P<0.05);肝衰竭组与肝癌组、肝硬化组、对照组CD8+比较明显升高(P值均<0.05);肝衰竭组较对照组、肝炎组、肝硬化组、肝癌组CD4+/CD8+明显降低(P值均<0.05);肝衰竭组、肝硬化组CD3-CD19+明显升高,分别与肝炎组、对照组比较差异均有统计学意义(P值均<0.05);肝衰竭组、肝癌组CD16+CD56+明显下降,分别与肝炎组、对照组比较差异均有统计学意义(P值均<0.05)。肝硬化组、肝衰竭组、肝癌组NF-κB、iNOS蛋白表达水平均明显增高,分别与肝炎组、对照组比较差异均有统计学意义(P值均<0.05)。与对照组比较,肝衰竭组、肝癌组、肝硬化组和肝炎组的NO和IL-6水平均明显升高(P值均<0.05)。结论 不同阶段HBV感染者免疫状态存在一定差异性,NF-κB-iNOS-NO可能参与HBV相关疾病的免疫反应。
【Abstract】:Objective To investigate the expression of nuclear factor-kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) in peripheral blood mononucleated cells (PBMCs) of patients with different stages of chronic hepatitis B (CHB), as well as the changes in natural killer (NK), T, and B lymphocytes in each stage. Methods A total of 80 patients with CHB-related diseases who were admitted to Nanping First Hospital Affiliated to Fujian Medical University from October 2017 to January 2019 were enrolled and divided into four groups according to disease stage: hepatitis group, liver failure group, liver cirrhosis group, and liver cancer group, with 20 patients in each group, and ten healthy volunteers were enrolled as control group. Flow cytometry was used to measure total lymphocyte count as well as the counts of lymphocyte subsets in peripheral blood in all five groups, Western blot was used to measure the protein expression of NF-κB and iNOS in PBMCs, and the nitrate reductase method and ELISA were used to measure the serum levels of nitric oxide (NO) and interleukin-6 (IL-6). A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the control group, the hepatitis group, the liver failure group, the liver cancer group, and the liver cirrhosis group had a significant reduction in total lymphocyte count (all P<0.05), and compared with the hepatitis group, the liver failure group, the liver cancer group, and the liver cirrhosis group had a significant reduction in total lymphocyte count (all P<0.05). The liver failure group, the liver cancer group, and the liver cirrhosis group had a significant reduction in CD3+ T cells compared with the control group and the hepatitis group (all P<0.05), as well as a significant reduction in CD4+ T cells compared with the hepatitis group (all P<0.05); the liver failure group had a significant increase in CD8+ T cells compared with the liver cancer group, the liver cirrhosis group, and the control group (all P<0.05); the liver failure group had a significant reduction in CD4+/CD8+ compared with the control group, the hepatitis group, the liver cirrhosis group, and the liver cancer group (all P<0.05); the liver failure group and the liver cirrhosis group had a significant increase in CD3-CD19+ B cells compared with the hepatitis group and the control group (all P<0.05); the liver failure group and the liver cancer group had a significant reduction in CD16+CD56+ NK cells compared with the hepatitis group and the control group (all P<0.05). The liver cirrhosis group, the liver failure group, and the liver cancer group had significant increases in the protein expression of NF-κB and iNOS compared with the hepatitis group and the control group (all P<0.05). Compared with the control group, the liver failure group, the liver cancer group, the liver cirrhosis group, and the hepatitis group had significant increases in the serum levels of NO and IL-6 (all P<0.05). Conclusion There are certain differences in immune status between patients with different stages of HBV infection, and NF-κB-iNOS-NO may be involved in the immune response of HBV-related diseases.
【关键字】:乙型肝炎,慢性; NF-κB; 一氧化氮合酶; 淋巴细胞
【Key words】:hepatitis B, chronic; NF-kappa B; nitric oxide synthase; lymphocytes
【引证本文】:HUANG J, ZHANG MQ, HE DX. Expression of nuclear factor-kappa B and inducible nitric oxide synthase in peripheral blood mononucleated cells and immune status in patients with different stages of chronic hepatitis B[J]. J Clin Hepatol, 2020, 36(3): 536-540. (in Chinese)
黄津, 张美泉, 何道兴. 不同阶段慢性乙型肝炎患者外周血单个核细胞中NF-κB/iNOS的表达及免疫状态[J]. 临床肝胆病杂志, 2020, 36(3): 536-540.

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