炎症性肠病患儿血清lncRNA THRIL和miR-495-3p表达与肠道常见菌种的相关性及其临床意义

doi:10.3969/j.issn.1673-8640.2025.06.009

摘要/Abstract

摘要：

目的探讨炎症性肠病（IBD）患儿血清长链非编码RNA（lncRNA）肿瘤坏死因子相关异种核糖核蛋白（THRIL）和miR-495-3p表达及其与肠道常见菌种的相关性，为IBD临床诊断提供参考。方法选取2021年6月—2023年5月郑州颐和医院儿科109例IBD患儿（研究组）和同期体检健康儿童109例（对照组）。采用培养方法鉴定入组儿童粪便样本中的肠道常见菌种，采用逆转录聚合酶链反应（PCR）检测血清lncRNA THRIL和miR-495-3p表达，通过ENCORI数据库预测lncRNA THRIL和miR-495-3p的结合位点。收集研究组和对照组相关临床资料，比较2组差异。采用Spearman相关分析评价IBD患儿血清lncRNA THRIL和miR-495-3p表达与肠道常见菌种菌落数的相关性。采用Logistic回归分析评估发生IBD的影响因素。采用受试者工作特征（ROC）曲线评价血清lncRNA THRIL和miR-495-3p诊断IBD的效能。结果研究组有肠道疾病史和不健康饮食习惯的患儿所占比例高于对照组（P<0.001）。研究组和对照组大肠埃希菌、双歧杆菌、乳酸杆菌、肠球菌、酵母菌菌落数差异均有统计学意义（P<0.001）；研究组血清lncRNA THRIL相对表达量高于对照组（P<0.001），miR-495-3p相对表达量低于对照组（P<0.001），且2项指标核酸序列存在互补位点；血清lncRNA THRIL相对表达量与大肠埃希菌、肠球菌、酵母菌菌落数呈正相关（r值分别为0.543、0.551、0.703，P<0.001），与双歧杆菌、乳酸杆菌菌落数呈负相关（r值分别为-0.561、-0.627，P<0.001）；miR-495-3p则相反。有肠胃疾病史[比值比（OR）值=1.772，P=0.029]、不健康饮食习惯（OR值=2.048，P=0.010）、lncRNA THRIL升高（OR值=12.117，P=0.001）和miR-495-3p降低（OR值=0.665，P=0.043）是IBD发生的危险因素。血清lncRNA THRIL和miR-495-3p联合检测诊断儿童IBD的效能优于2项指标单独检测（P<0.001）。结论血清lncRNA THRIL和miR-495-3p相对表达量与IBD患儿肠道常见菌种密切相关，2项指标检测对IBD具有较高的诊断价值。

关键词: 长链非编码RNA, 肿瘤坏死因子相关异种核糖核蛋白, 微小RNA-495-3p, 肠道常见菌种, 炎症性肠病, 儿童

Abstract:

Objective To investigate serum long non-coding RNA（lncRNA） tumor necrosis factor-related heterogeneous ribonucleoprotein particle（THRIL） and miR-495-3p expressions in children with inflammatory bowel disease（IBD） and their correlation with common intestinal bacterial species，and to provide a reference for the clinical diagnosis of IBD. Methods From June 2021 to May 2023，109 children with IBD admitted to Zhengzhou Yihe Hospital were enrolled as study group，and another 109 healthy children were enrolled as control group. The common intestinal bacterial species in the fecal samples of enrolled children were identified by culture methods. The expression levels of serum lncRNA THRIL and miR-495-3p were determined by reverse transcription polymerase chain reaction（PCR）. Using ENCORI database，the binding site between lncRNA THRIL and miR-495-3p was predicted. The relevant clinical data of study group and control group were collected，and the differences between the 2 groups were compared. Spearman method was used to analyze the correlation between the expression levels of serum lncRNA THRIL and miR-495-3p and the number of common intestinal bacterial species. Logistic regression analysis was used to analyze the related factors for the occurrence of IBD. The diagnostic value of serum lncRNA THRIL and miR-495-3p for IBD was analyzed by receiver operating characteristic（ROC） curve. Results The proportion of patients with a history of intestinal diseases and unhealthy dietary habits in study group was higher than that in control group（P<0.01）. There was statistical significance in the colony numbers of Escherichia coli，Bifidobacterium，Lactobacillus，Enterococcus and yeast between the 2 groups（P<0.001）. The relative expression level of serum lncRNA THRIL in study group was higher than that in control group（P<0.001），while the relative expression level of miR-495-3p was lower than that in control group（P<0.001），and there were complementary sites in the nucleic acid sequences of the 2 indicators. Serum lncRNA THRIL relative expression level was positively correlated with the colony numbers of Escherichia coli，Enterococcus and yeast（r=0.543，0.551 and 0.703，P<0.001），and it was negatively correlated with the colony numbers of Bifidobacterium and Lactobacillus（r=-0.561 and -0.627，P<0.001）. The miR-495-3p was on the contrary. History of gastrointestinal diseases [odds ratio（OR）=1.772，P=0.029]，unhealthy dietary habits（OR=2.048，P=0.010），increased lncRNA THRIL（OR=12.117，P=0.001） and decreased miR-495-3p（OR=0.665，P=0.043） were risk factors for the occurrence of IBD. The combined determination of serum lncRNA THRIL and miR-495-3p was superior to the single determinations（P<0.001）. Conclusions The relative expression levels of serum lncRNA THRIL and miR-495-3p are related to the common intestinal bacterial species of patients with IBD. The combined determination has a high diagnostic value for IBD.

Key words: Long non-coding RNA, Tumor necrosis factor-related heterogeneous ribonucleoprotein particle, MicroRNA-495-3p, Common intestinal bacterial sepecies, Inflammatory bowel disease, Children

中图分类号:

R446.7

燕华, 高大汉, 刘东锋. 炎症性肠病患儿血清lncRNA THRIL和miR-495-3p表达与肠道常见菌种的相关性及其临床意义[J]. 检验医学, 2025, 40(6): 565-570.

YAN Hua, GAO Dahan, LIU Dongfeng. Correlation and clinical significance of serum lncRNA THRIL and miR-495-3p expressions with common intestinal bacterial species in children with inflammatory bowel disease[J]. Laboratory Medicine, 2025, 40(6): 565-570.

图/表 8

参考文献 23

[1]	邢瑞欣, 王洪乾, 章文慧, 等. 检测炎症性肠病患者血清铜/锌比值的临床意义[J]. 中华消化杂志, 2021, 41(10):665-670.
[2]	王宏刚, 姜建国, 王勇. 饮食在炎症性肠病发病机制中的作用[J]. 中华炎性肠病杂志, 2020, 4(2):146-149.
[3]	KELSEN J R, RUSSO P, SULLIVAN K E. Early-onset inflammatory bowel disease[J]. Immunol Allergy Clin North Am, 2019, 39(1):63-79.
[4]	CHAPARRO M, GARRE A, RICART E, et al. Short and long-term effectiveness and safety of vedolizumab in inflammatory bowel disease:results from the ENEIDA registry[J]. Aliment Pharmacol Ther, 2018, 48(8):839-851.
[5]	WDRYCHOWICZ A, TOMASIK P, ZAJC A, et al. Prognostic value of assessment of stool and serum IL^-1β,IL^-1ra and IL-6 concentrations in children with active and inactive ulcerative colitis[J]. Arch Med Sci, 2018, 14(1):107-114.
[6]	苏婧玲, 谢晨曦, 范燕云, 等. IL^-17BR在炎症性肠病中的表达及其临床意义[J]. 胃肠病学, 2018, 23(2):83-87.
[7]	杨默媛, 安泽, 孙宇涵, 等. 枯草杆菌二联活菌肠溶胶囊联合泮托拉唑对UC患者炎性因子、肠黏膜功能及外周血Th17、CD4+CD25+Treg细胞表达的影响[J]. 现代生物医学进展, 2020, 20(20):3965-3968.
[8]	刘巍. 中英夫利昔单抗联合中药治疗炎症性肠病活动期的临床疗效及对免疫功能的影响分析[J]. 河北医学, 2020, 26(1):150-154.
[9]	NAF A, OGS B, EAM C, et al. Diagnostic potential of metastasis associated lung adenocarcinoma transcript-1(MALAT-1)and TNFα and hnRNPL related immunoregulatory long non-coding RNA(THRIL)in systemic lupus erythematosus patients:relation to disease activity[J]. Egypt Rhcumatolo, 2019, 41(3):197-201.
[10]	SAYAD A, HAJIFATHALI A, OMRANI M D, et al. Expression of TNF- and HNRNPL-related Immunoregulatory long non-coding RNA(THRIL)in acute myeloid leukemia:is there any correlation?[J]. Iran J Allergy Asthma Immunol, 2018, 17(3):274-280.
[11]	杨文宏, 孙惠芳, 杨洁. MiR-125b和miR-335在炎症性肠病患者外周血和肠黏膜中的表达研究[J]. 重庆医学, 2021, 50(5):800-805.
[12]	崔昭, 梁博, 李国东, 等. 炎症性肠病患者血清miR-181a-5p和miR-126表达水平及其与肠道菌群相关性的研究[J]. 国际消化病杂志, 2021, 41(1):33-37.
[13]	中华医学会消化病学分会炎症性肠病学组. 炎症性肠病诊断与治疗的共识意见(2018 年,北京)[J]. 中华消化杂志, 2018, 38(5):292-311.
[14]	BEARD J A, FRANCO D L, CLICK B H. The burden of cost in inflammatory bowel disease:a medical economic perspective and the future of value-based care[J]. Curr Gastroenterol Rep, 2020, 22(2):6.
[15]	LLOYD-PRICE J, ARZE C, ANANTHAKRISHNAN A N, et al. Multi-omics of the gut microbial ecosystem in inflammatory bowel diseases[J]. Nature, 2019, 569(7758):655-662.
[16]	石钰洁, 羽思, 田博文, 等. 炎症性肠病患者伴发泌尿系结石的相关因素及肠道菌群和代谢物特征分析[J]. 中华医学杂志, 2022, 102(44):3525-3531.
[17]	杨志勇, 刘维娜. LncRNA THRIL在ICU脓毒症患者血清中的表达水平及临床价值[J]. 检验医学与临床, 2023, 20(1):109-111.
[18]	王蒲春, 刘德志, 范新, 等. 血浆长链非编码RNA THRIL、NEAT1表达水平与脓毒症并发急性呼吸窘迫综合征的关系[J]. 临床急诊杂志, 2021, 22(8):534-540.
[19]	ELAMIR A, SHAKER O, KAMAL M, et al. Expression profile of serum lncRNA THRIL and miR-125b in inflammatory bowel disease[J]. PLoS One, 2022, 17(10):e0275267.
[20]	SONG X, GAO F, LI H, et al. Long noncoding RNA THRIL promotes foam cell formation and inflammation in macrophages[J]. Cell Biol Int, 2023, 47(1):156-166. DOI PMID
[21]	DENG W, WANG X, ZHANG J, et al. Circ_0138959/miR-495-3p/TRAF6 axis regulates proliferation,wound healing and osteoblastic differentiation of periodontal ligament cells in periodontitis[J]. J Dent Sci, 2022, 17(3):1125-1134.
[22]	SONG W, QIU N. MiR-495-3p depletion contributes to myocardial ischemia/reperfusion injury in cardiomyocytes by targeting TNC[J]. Regen Ther, 2022, 21:380-388. DOI PMID
[23]	ZHANG J L, ZHENG H F, LI K, et al. MiR-495-3p depresses cell proliferation and migration by downregulating HMGB1 in colorectal cancer[J]. World J Surg Oncol, 2022, 20(1):101.

引物名称	正向引物（5'~3'）	反向引物（5'~3'）
lncRNA THRIL	AACTTCACAGGAACACTACACAAGA	TAGGCAACAGAGAGCAAGACTTCATC
miR-495-3p	GCGGAAACAAACATGGTGCA	CAGTGCGTGTCGTGGAGT
GAPDH	GCACCCTCAAGGCTGAGAAC	TGGTGAAGACGCCAGTGGA
U6	ATTGGAACGATACAGAGAAGATT	GGAACGCTTCACGAATTTG

引物名称	正向引物（5'~3'）	反向引物（5'~3'）
lncRNA THRIL	AACTTCACAGGAACACTACACAAGA	TAGGCAACAGAGAGCAAGACTTCATC
miR-495-3p	GCGGAAACAAACATGGTGCA	CAGTGCGTGTCGTGGAGT
GAPDH	GCACCCTCAAGGCTGAGAAC	TGGTGAAGACGCCAGTGGA
U6	ATTGGAACGATACAGAGAAGATT	GGAACGCTTCACGAATTTG

组别	年龄		性别		体重指数		营养不良
组别	<8岁	≥8岁	男	女	<22 kg·m^-2	≥22 kg·m^-2	是	否
对照组	53.95（41/76）	47.89（68/142）	46.02（52/113）	54.29（57/105）	47.00（47/100）	52.54（62/118）	52.73（87/165）	41.51（22/53）
研究组	46.05（35/76）	52.11（74/142）	53.98（61/113）	45.71（48/105）	53.00（53/100）	47.46（56/118）	47.27（78/165）	58.49（31/53）
χ²值	0.727		1.488		0.665		2.019
P值	0.394		0.222		0.415		0.155
组别	家族IBD疾病史			肠胃疾病史			不健康饮食习惯
组别	有	无		有	无		有	无
对照组	42.62（26/61）	52.87（83/157）		37.93（44/116）	63.73（65/102）		38.61（39/101）	59.83（70/117）
研究组	57.38（35/61）	47.13（74/157）		62.07（72/116）	36.27（37/102）		61.39（62/101）	40.17（47/117）
χ²值	1.844			14.445			9.759
P值	0.175			<0.001			0.002

组别	年龄		性别		体重指数		营养不良
组别	<8岁	≥8岁	男	女	<22 kg·m^-2	≥22 kg·m^-2	是	否
对照组	53.95（41/76）	47.89（68/142）	46.02（52/113）	54.29（57/105）	47.00（47/100）	52.54（62/118）	52.73（87/165）	41.51（22/53）
研究组	46.05（35/76）	52.11（74/142）	53.98（61/113）	45.71（48/105）	53.00（53/100）	47.46（56/118）	47.27（78/165）	58.49（31/53）
χ²值	0.727		1.488		0.665		2.019
P值	0.394		0.222		0.415		0.155
组别	家族IBD疾病史			肠胃疾病史			不健康饮食习惯
组别	有	无		有	无		有	无
对照组	42.62（26/61）	52.87（83/157）		37.93（44/116）	63.73（65/102）		38.61（39/101）	59.83（70/117）
研究组	57.38（35/61）	47.13（74/157）		62.07（72/116）	36.27（37/102）		61.39（62/101）	40.17（47/117）
χ²值	1.844			14.445			9.759
P值	0.175			<0.001			0.002

组别	例数	大肠埃希菌/ （lgCFU·g^-1）	双歧杆菌/ （lgCFU·g^-1）	乳酸杆菌/ （lgCFU·g^-1）	肠球菌/ （lgCFU·g^-1）	葡萄球菌/ （lgCFU·g^-1）	酵母菌/ （lgCFU·g^-1）
对照组	109	5.72±1.03	6.41±1.21	5.26±1.08	5.08±0.92	4.36±0.79	2.03±0.35
研究组	109	6.33±1.27	4.61±0.96	3.84±0.72	7.06±1.52	4.53±0.86	2.35±0.49
t值		3.895	12.167	11.422	11.635	1.520	5.548
P值		<0.001	<0.001	<0.001	<0.001	0.130	<0.001