不同疾病活动度炎症性肠病患者血清microRNA-146a-5p、microRNA-145水平的变化及与免疫调节的相关性

Changes in serum levels of miRNA-146a-5p and miRNA-145 in patients with different disease activity levels of inflammatory bowel disease and their correlation with immune regulation

摘要:

目的 探讨不同疾病活动度炎症性肠病（IBD）患者血清microRNA-146a-5p（miR-146a-5p）、microRNA-145（miR-145）水平的变化及与免疫调节的相关性。方法 选取2021年6月—2022年6月在西南医科大学附属中医医院就诊的88例IBD患者作为IBD组，其中溃疡性结肠炎（UC）43例，克罗恩病（CD）45例。另取同期该院86例健康体检者作为对照组。采用实时荧光定量聚合酶链反应检测血清和肠黏膜组织miR-146a-5p、miR-145表达，流式细胞术检测Th1、Th2，酶联免疫吸附试验测定血清肿瘤坏死因子-α（TNF-α）、白细胞介素-4（IL-4）、干扰素-γ（IFN-γ）、白细胞介素-12（IL-12）水平。Pearson法分析患者miR-146a-5p、miR-145表达与Th1/Th2的相关性。结果 与对照组比较，UC组和CD组miR-145相对表达量降低（P <0.05），Th1、Th1/Th2升高（P <0.05）；与UC组比较，CD组miR-145相对表达量降低（P <0.05），Th1、Th1/Th2升高（P <0.05）。对照组、UC组、CD组miR-146a-5p都在固有层中表达。UC组和CD组miR-146a-5p表达水平高于对照组（P <0.05）。UC组和CD组TNF-α、IFN-γ、IL-4、IL-12水平高于对照组（P <0.05）。Pearson相关性分析结果表明，miR-146a-5p、miR-145表达与Th1/Th2均呈负相关（r =-0.424和-0.395，均P =0.000）。miR-146a-5p、miR-145诊断UC的曲线下面积（AUC）分别为0.850和0.794，敏感性分别为79.5%（95% CI：0.718，0.857）和74.4%（95% CI：0.696，0.822），特异性分别为94.3%（95% CI：0.982，0.896）和72.5%（95% CI：0.679，0.773）。miR-146a-5p、miR-145诊断CD的AUC分别为0.927（95% CI：0.875，0.977）和0.846（95% CI：0.803，0.902），敏感性分别为92.3%（95% CI：0.875，0.978）和71.9%（95% CI：0.668，0.773），特异性分别为94.3%（95% CI：0.991，0.892）和90.9%（95% CI：0.945，0.847）。miR-146a-5p、miR-145对CD的诊断效能较高。结论 不同疾病活动度IBD患者血清miR-146a-5p、miR-145表达与免疫调节呈负相关。

Abstract:

Objective To investigate the changes in serum levels of miRNA-146a-5p and miRNA-145 in patients with different disease activity levels of inflammatory bowel disease (IBD) and their correlation with immune regulation.Methods A total of 88 IBD patients who were treated in the Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University from June 2021 to June 2022 were selected as the IBD group, including 43 patients with ulcerative colitis (UC group) and 45 patients with Crohn's disease (CD group). Meanwhile, 86 healthy people undergoing health checkup in the hospital during the same period were selected as the control group. The expression levels of miRNA-146a-5p and miRNA-145 in serum and intestinal mucosa were detected by quantitative real-time polymerase chain reaction. The frequency of Th1 and Th2 cells was measured by flow cytometry. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-12 (IL-12) were determined by enzyme-linked immunosorbent assay. Pearson method was applied to analyze the correlation between the expressions of miRNA-146a-5p and miRNA-145 and the Th1/Th2 ratio.Results Compared with the control group, the relative expression level of miR-145 was lower (P < 0.05) but the frequency of Th1 cells and the Th1/Th2 ratio were higher (P < 0.05) in the UC group and the CD group. Compared with the UC group, the relative expression level of miR-145 was lower (P < 0.05) but the frequency of Th1 cells and the Th1/Th2 ratio were higher (P < 0.05) in the CD group. In all groups, miR-146a-5p was expressed in the lamina propria. The expression levels of miR-146a-5p in the UC group and the CD group were higher than those in the control group (P < 0.05). The levels of TNF-α, IFN-γ, IL-4 and IL-12 in the UC group and the CD group were also higher than those in the control group (P < 0.05). Pearson correlation analysis demonstrated that the expressions of miR-146a-5p and miR-145 were both negatively correlated with the Th1/Th2 ratio (r = -0.424 and -0.395, both P = 0.000). The areas under the curves (AUCs) of miR-146a-5p and miR-145 for diagnosing UC were 0.850 and 0.794, with sensitivities being 79.5% (95% CI: 0.718, 0.857) and 74.4% (95% CI: 0.696, 0.822), and specificities being 94.3% (95% CI: 0.982, 0.896) and 72.5% (95% CI: 0.679, 0.773). The AUCs of miR-146a-5p and miR-145 for diagnosing CD were 0.927 and 0.846, with the sensitivities being 92.3% (95% CI: 0.875, 0.978) and 71.9% (95% CI: 0.668, 0.773), and the specificities being 94.3% (95% CI: 0.991, 0.892) and 90.9% (95% CI: 0.945, 0.847), indicating that miR-146a-5p and miR-145 exhibited higher diagnostic efficacy for CD.Conclusions The serum expression levels of miR-146a-5p and miR-145 are negatively correlated with immune regulation in patients with different disease activity levels of IBD.