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T、B淋巴细胞自噬在自身免疫性肝炎患者外周血中的表达及临床意义
DOI: 10.3969/j.issn.1001-5256.2022.11.009
伦理学声明：本研究方案于2019年3月25日经由首都医科大学附属北京佑安医院伦理委员会审批，批号：LL-2019-184-K。
利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。
作者贡献声明：伍慧丽、时红波、刘燕敏负责课题设计，资料分析，撰写论文；刘燕敏、严艳、王玲玲、丁美、马正来、刘晖参与收集数据，修改论文；段钟平负责拟定写作思路，指导撰写文章并最后定稿。
Expression of autophagy marker in peripheral blood T and B lymphocytes of patients with autoimmune hepatitis and its clinical significance
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摘要:
目的 探讨自身免疫性肝炎(AIH)患者外周血T、B淋巴细胞自噬表达及其临床意义。 方法 选取2019年10月—2020年10月在首都医科大学附属北京佑安医院门诊或住院治疗的62例AIH患者及8例健康对照者外周血进行T、B淋巴细胞亚群自噬相关检测，根据治疗情况、诊断类型、是否合并肝硬化及肝衰竭进行分组分析。符合正态分布的计量资料两组间比较采用t检验，非正态分布计量资料多组间比较采用Kruskal-Wallis H检验，两组间比较采用Mann-Whitney U检验；计数资料组间比较采用χ2检验或Fisher精确概率法。 结果 AIH组CD4+T、CD8+T、CD19+B及CD4+CD25+T淋巴细胞自噬LC3B平均荧光强度(MFI)均显著高于健康对照组(P值均＜0.05)，以CD19+B淋巴细胞自噬LC3B MFI最显著。CD19+B淋巴细胞自噬MFI在未治疗组和治疗部分缓解组中高于治疗完全缓解组(P值分别为0.037、0.040)，在特发性AIH组(I-AIH)和药物诱导的AIH组(DI-AIH)高于重叠综合征组(PBC-AIH)(P值分别为0.037、0.031)，在无肝硬化组和失代偿期肝硬化组高于代偿期肝硬化组(P值分别为0.009、0.003)，在肝衰竭组中高于无肝衰竭组(P=0.042)；CD4+CD25+T淋巴细胞自噬MFI在PBC-AIH组高于I-AIH组和DI-AIH组(P值分别为0.042、0.044)，在代偿期肝硬化组低于无肝硬化组(P=0.037)，无肝衰竭组中高于肝衰竭组(P=0.043)。 结论 AIH患者较健康人外周血T、B淋巴细胞亚群自噬表达升高，其自噬水平与治疗、诊断类型、疾病严重程度有关。 Abstract:Objective To investigate the expression of autophagy marker in peripheral blood T and B lymphocytes of patients with autoimmune hepatitis (AIH) and its clinical significance. Methods Peripheral blood samples were collected from 62 AIH patients who were treated in Beijing YouAn Hospital affiliated to Capital Medical University from October 2019 to October 2020 who were treated in Beijing YouAn Hospital affiliated to Capital Medical University from October 2019 to October 2020 and 8 healthy controls to detect autophagy of T and B lymphocyte subsets, and then subgroup analyses were performed based on treatment, diagnostic type, and presence or absence of liver cirrhosis and liver failure. The t-test was used for comparison of normally distributed continuous data between two groups; the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Mann-Whitney U test was used for comparison between two groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. Results Compared with the healthy control group, the AIH group had a significantly higher mean fluorescence intensity (MFI) of the autophagy marker LC3B in CD4+ T, CD8+ T, CD19+ B, and CD4+CD25+ T lymphocytes (all P < 0.05), especially in CD19+ B lymphocytes. The non-treatment group and the partial remission group had a significantly higher MFI of autophagy marker in CD19+ B lymphocytes than the complete remission group (P=0.037 and 0.040); the idiopathic AIH (I-AIH) group and the drug-induced AIH(DI-AIH) group had a significantly higher MFI than the primary biliary cholangitis (PBC)-AIH overlap syndrome group (P=0.037 and 0.031); the non-cirrhosis group and the decompensated cirrhosis group had a significantly higher MFI than the compensated cirrhosis group (P=0.009 and 0.003); the liver failure group had a significantly higher MFI than the non-liver failure group (P=0.042). The PBC-AIH group had a significantly higher MFI of autophagy marker in CD4+CD25+ T lymphocytes than the I-AIH group and the DI-AIH group (P=0.042 and 0.044), the compensated cirrhosis group had a significantly lower MFI than the non-cirrhosis group (P=0.037), and the non-liver failure group had a significantly higher MFI than the liver failure group (P=0.043). Conclusion AIH patients have a significant increase in the expression of autophagy marker in peripheral blood T and B lymphocyte subsets compared with healthy individuals, and the level of autophagy is associated with treatment, diagnostic type, and disease severity. -
Key words:
- Hepatitis, Autoimmune /
- Lymphocytes /
- Autophagy
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表 1 AIH患者临床特点
Table 1. Clinical characteristics of AIH patients
指标 数值 诊断类型[例(%)] I-AIH 41(66.1) DI-AIH 11(17.7) PBC-AIH 10(16.1) 肝衰竭[例(%)] 急/亚急性肝衰竭 4(6.5) 慢加急/亚急性肝衰竭 4(6.5) 慢性肝衰竭 4(6.5) 肝衰竭分期[例(%)] 早期(30%＜PTA≤40%) 9(14.5) 中期(20%＜PTA≤30%) 2(3.2) 晚期(PTA≤20%) 1(1.6) 皮质醇激素或免疫抑制剂治疗[例(%)] 皮质醇激素 21(33.9) 皮质醇激素+免疫抑制剂 9(14.5) 临床症状[例(%)] 纳差 31(50.0) 尿黄 32(51.6) 腹胀 15(24.2) 水肿 8(12.9) 恶心、呕吐 11(17.7) 发热 5(8.1) 无症状 10(16.1) 肝硬化[例(%)] 34(54.8) 糖尿病[例(%)] 10(16.1) 合并其他自身免疫疾病[例(%)] 15(24.2) 接受病理检查[例(%)] 42(67.7) 
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表 2 AIH组与健康对照组T、B淋巴细胞亚群自噬MFI表达
Table 2. Autophagic MFI expression of T and B lymphocyte subsets in AIH group and healthy control group
指标 AIH组(n=62) 健康对照组(n=8) Z值 P值 CD4+LC3B+T 2238(1664~2810) 1237(616~1392) 2.254 0.024 CD8+LC3B+T 2309(1336~2806) 1367(421~1557) 2.288 0.022 CD19+LC3B+B 31 861(9375~42 799) 1195(764~1303) 2.380 0.017 CD4+CD25+LC3B+T 2959(1331~3988) 675(353~924) 2.288 0.022
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表 3 AIH患者糖皮质激素或免疫抑制剂治疗前后T、B淋巴细胞亚群自噬表达
Table 3. Autophagy expression of T and B lymphocyte subsets in AIH patients before and after glucocorticoid or immunosuppressant treatment
组别 例数 CD4+T LC3B MFI CD8+T LC3B MFI CD19+B LC3B MFI CD4+CD25+ LC3B MFI 健康对照组 8 1237(616~1392) 1367(421~1557) 1195(764~1303) 675(353~924) 未治疗组 32 2106(1391~2639)2) 2289(1339~3104)2) 32 522(10 841~48 884)1)3) 2704(1208~4163)1) 完全缓解组 13 2043(1608~2731)1) 2478(1327~2784) 16 390(4980~20 029)1) 3264(1772~3349)2) 部分缓解组 17 2813(1760~3060)2) 1550(1267~2632) 32 501(1146~50 043)1)3) 1480(1083~5242)2) χ2值 25.474 16.191 28.085 23.007 P值 ＜0.001 0.001 ＜0.001 ＜0.001 注：与健康对照组比较，1) P＜0.01, 2)P＜0.05；与完全缓解组比较，3)P＜0.05。
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表 4 不同AIH诊断类型与T、B淋巴细胞亚群自噬表达
Table 4. Different diagnostic types of AIH and autophagy expression of T and B lymphocyte subsets
组别 例数 CD4+T LC3B MFI CD8+T LC3B MFI CD19+B LC3B MFI CD4+CD25+ LC3B MFI 健康对照组 8 1237(616~1392) 1367(421~1557) 1195(764~1303) 675(353~924) I-AIH组 41 1982(1516~2810)1) 1782(1194~2394) 32 511(8933~48 625)1)3) 2585(1095~3153)1)3) DI-AIH组 11 2214(1831~3167)1) 2194(1696~2736)1) 37 148(12 301~81 896)1)3) 2575(2180~2994)1)3) PBC-AIH组 10 1960(864~2075) 1798(950~1748) 17 956(763~26 171) 4648(2123~5739)2) χ2值 24.913 17.362 30.736 26.409 P值 ＜0.001 0.001 ＜0.001 ＜0.001 注：与健康对照组比较，1) P＜0.01, 2)P＜0.05；与PBC-AIH组比较，3) P＜0.05。
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表 5 AIH患者肝硬化情况与淋巴细胞亚群自噬表达
Table 5. Autophagy expression of lymphocyte subsets in AIH patients with different liver cirrhosis
组别 例数 CD4+T LC3B MFI CD8+T LC3B MFI CD19+B LC3B MFI CD4+CD25+ LC3B MFI 健康对照组 8 1237(616~1392) 1367(421~1557) 1195(764~1303) 675(353~924) 无肝硬化组 28 2096(1706~2964)1) 2422(1551~2670)1) 33 284(12 754~42 799)1)3) 2959(2208~3468)1)3) 代偿期肝硬化组 14 1921(1391~2147)2) 1498(1267~2015) 1357(1146~34 267) 1480(1083~3073)2) 失代偿期肝硬化组 20 2639(1367~2936)2) 1789(1108~2319) 32 522(13 888~56 323)1)3) 1772(1120~5025)2) χ2值 26.766 19.573 34.762 26.657 P值 ＜0.001 ＜0.001 ＜0.001 ＜0.001 注：与健康对照组比较，1) P＜0.01, 2)P＜0.05；与代偿期肝硬化组比较，3) P＜0.05。
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表 6 AIH患者肝衰竭情况与淋巴细胞亚群自噬表达之间的关系
Table 6. Autophagy expression of lymphocyte subsets in AIH patients with different liver failure
组别 例数 CD4+T LC3B MFI CD8+T LC3B MFI CD19+B LC3B MFI CD4+CD25+ LC3B MFI 健康对照组 8 1237(616~1392) 1367(421~1557) 1195(764~1303) 675(353~924) 无肝衰竭组 50 2491(1664~2824)1) 2367(1336~2806)2) 18442(9375~40 275)1) 3147(1699~4081)1) 有肝衰竭组 12 1921(1495~2457)2) 1550(1034~2931) 35 346(16 823~50 644)2)3) 1372(541~3335)3) χ2值 25.520 16.188 29.431 26.035 P值 ＜0.001 ＜0.001 ＜0.001 ＜0.001 注：与健康对照组比较，1) P＜0.01, 2)P＜0.05；与无肝衰竭组比较，3)P＜0.05。
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