老年维持性血透患者叶酸治疗与miR-150-5p血清水平的相关性研究

doi:10.3877/cma.j.issn.2095-3216.2023.03.004

目的

探讨老年维持性血透患者服用叶酸与微小RNA-150-5p(miR-150-5p)血清水平的关系。

方法

2020年1月至2021年1月于首都医科大学附属北京友谊医院医疗保健中心收治的老年维持性血液透析(MHD)患者28例，根据是否服用叶酸治疗分为用药组(12例)和对照组(16例)。比较两组的基线资料、用药前后叶酸和肿瘤标志物水平。实时定量PCR检测血清miR-150-5p水平，评估叶酸水平与miR-150-5p水平、肿瘤标志物之间的相关性。利用生物信息学和KEGG模型分析miR-150-5p可能参与的信息通路。

结果

与对照组相比，用药组的叶酸水平明显增高，而癌胚抗原(CEA)、癌抗原125(CA125)含量以及miR-150-5p表达量降低(P<0.05)。与用药前相比，用药后的叶酸水平显著上升(P<0.001)，而miR-150-5p表达量则明显下调(P<0.05)。Pearson相关性分析提示血清叶酸与miR-150-5p呈负相关(P<0.05)。生物信息学分析提示miR-150-5p的下游靶基因多与肿瘤信号通路相关。

结论

老年MHD患者服用叶酸后，miR-150-5p和CEA水平均有下降。叶酸与miR-150-5p呈负相关，而miR-150-5p的下游靶基因多与肿瘤信号通路相关，提示通过叶酸调控miR-150-5p表达，有可能影响肿瘤的发生发展。

关键词: 维持性血液透析, 老年, 叶酸, 微小RNA, 肿瘤生物标志物

Objective

To investigate the relationship between folic acid intake and serum level of miR-150-5p in elderly patients undergoing maintenance hemodialysis.

Methods

From January 2020 to January 2021, 28 elderly patients undergoing maintenance hemodialysis in the Beijing Friendship Hospital Medical Care Center were divided into a treatment group (n=12) with folic acid therapy and a control group (n=16) without folic acid therapy. The baseline data, serum folic acid and tumor markers were compared. Real-time PCR was used to analyze the serum level of miR-150-5p. The correlation among the folic acid, miR-150-5p, and tumor markers was analyzed. Bioinformatics and KEGG model were used to analyze potential regulatory pathways of miR-150-5p.

Results

Compared with the control group, the treatment group showed higher serum level of folic acid, but lower serum levels of CEA, CA125, and miR-150-5p (P<0.01). In comparison with those before the treatment, folic acid level was significantly increased after treatment (P<0.001), while miR-150-5p was significantly down-regulated (P<0.05). Pearson correlation analysis showed that serum folic acid level was negatively correlated with the serum level of miR-150-5p (P<0.05), and bioinformatics analysis suggested that most of the downstream target genes of miR-150-5p were involved in tumor signaling pathways.

Conclusion

After receiving the folic acid therapy, the elderly patients with maintenance hemodialysis had lower serum levels of miR-150-5p and CEA decreased. The serum level of folic acid was negatively correlated with miR-150-5p, while the downstream target genes of miR-150-5p were mostly involved in tumor signaling pathways, suggesting that regulating miR-150-5p expression through folic acid might have a role to affect the occurrence and development of tumors.

Key words: Maintenance hemodialysis, Old age, Folic acid, microRNA, Tumor biomarkers

表2 用药组和对照组一般情况对比

组别	全体患者(n＝28)	对照组(n＝16)	用药组(n＝12)	t值	P值
一般资料
男性[例(%)]	16(57.0)	9(56.3)	7(58.3)	0.239	0.735
年龄(岁)	78.8±5.1	79.3±4.7	78.1±5.6	0.276	0.708
身体质量指数(kg/m²)	23.7±3.6	23.4±2.6	24.2±3.7	－0.593	0.608
收缩压(mmHg)	145.2±16.8	146.1±17.4	143.2±15.3	0.873	0.322
舒张压(mmHg)	75.6±11.3	76.5±14.2	74.2±7.8	0.731	0.482
基础疾病史[例(%)]
冠心病	14(50.0)	6(37.5)	8(66.7)	－1.459	0.067
高血压	22(78.6)	13(81.3)	9(75.0)	0.679	0.523
糖尿病	10(35.7)	6(37.5)	4(33.3)	0.223	0.793
透析时间(年)	4.9±3.3	5.4±3.3	4.1±3.6	0.915	0.258
化验情况
白细胞(10⁹/L)	5.82±1.1	6.1±0.4	5.6±1.5	0.683	0.523
中性粒细胞百分比(%)	66.7±8.1	64.8±8.0	68.6±8.4	－0.762	0.455
血红蛋白(g/L)	118.1±12.2	119.1±10.9	117±14.4	0.432	0.775
白蛋白(g/dl)	39.1±4.8	38.1±2.7	39.9±6.4	－0.603	0.530
eGFR[ml/(min·1.73 m²)]	6.4±3.1	5.8±3.7	6.9±2.9	－1.245	0.249
尿素氮(mg/dL)	20.2±3.8	21.4±3.6	18.9±3.7	1.164	0.267

表2 用药组和对照组一般情况对比

组别	全体患者(n＝28)	对照组(n＝16)	用药组(n＝12)	t值	P值
一般资料
男性[例(%)]	16(57.0)	9(56.3)	7(58.3)	0.239	0.735
年龄(岁)	78.8±5.1	79.3±4.7	78.1±5.6	0.276	0.708
身体质量指数(kg/m²)	23.7±3.6	23.4±2.6	24.2±3.7	－0.593	0.608
收缩压(mmHg)	145.2±16.8	146.1±17.4	143.2±15.3	0.873	0.322
舒张压(mmHg)	75.6±11.3	76.5±14.2	74.2±7.8	0.731	0.482
基础疾病史[例(%)]
冠心病	14(50.0)	6(37.5)	8(66.7)	－1.459	0.067
高血压	22(78.6)	13(81.3)	9(75.0)	0.679	0.523
糖尿病	10(35.7)	6(37.5)	4(33.3)	0.223	0.793
透析时间(年)	4.9±3.3	5.4±3.3	4.1±3.6	0.915	0.258
化验情况
白细胞(10⁹/L)	5.82±1.1	6.1±0.4	5.6±1.5	0.683	0.523
中性粒细胞百分比(%)	66.7±8.1	64.8±8.0	68.6±8.4	－0.762	0.455
血红蛋白(g/L)	118.1±12.2	119.1±10.9	117±14.4	0.432	0.775
白蛋白(g/dl)	39.1±4.8	38.1±2.7	39.9±6.4	－0.603	0.530
eGFR[ml/(min·1.73 m²)]	6.4±3.1	5.8±3.7	6.9±2.9	－1.245	0.249
尿素氮(mg/dL)	20.2±3.8	21.4±3.6	18.9±3.7	1.164	0.267

表1 PCR引物序列情况

引物名称	引物序列(5′-3′)
hsa-miR-150-5p-R	TGCCAGGCTGAAGGATCACT
hsa-miR-150-5p-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCACTGG
U6-R	GCGCGTCGTGAAGCGTTC
U6-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAAATATG

表1 PCR引物序列情况

引物名称	引物序列(5′-3′)
hsa-miR-150-5p-R	TGCCAGGCTGAAGGATCACT
hsa-miR-150-5p-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCACTGG
U6-R	GCGCGTCGTGAAGCGTTC
U6-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAAATATG

表3 两组患者用药前后叶酸水平和血清中miR-150-5p相对表达量比较(±s)

组别	叶酸(ng/ml)				miR-150-5p相对表达量
组别	用药前	用药后	t值	P值	用药前	用药后	t值	P值
对照组(n＝16)	4.1±1.9	4.8±0.9	－0.711	0.428	1.52±0.34	1.47±0.24	0.633	0.525
用药组(n＝12)	3.5±2.6	15.5±7.2^ab	－7.143	<0.001	1.53±0.46	1.23±0.28^ab	2.849	0.016
t值	0.793	－4.649			－0.319	2.968
P值	0.546	<0.001			0.772	0.007

表3 两组患者用药前后叶酸水平和血清中miR-150-5p相对表达量比较(±s)

组别	叶酸(ng/ml)				miR-150-5p相对表达量
组别	用药前	用药后	t值	P值	用药前	用药后	t值	P值
对照组(n＝16)	4.1±1.9	4.8±0.9	－0.711	0.428	1.52±0.34	1.47±0.24	0.633	0.525
用药组(n＝12)	3.5±2.6	15.5±7.2^ab	－7.143	<0.001	1.53±0.46	1.23±0.28^ab	2.849	0.016
t值	0.793	－4.649			－0.319	2.968
P值	0.546	<0.001			0.772	0.007

表4 两组患者用药前后肿瘤标志物比较(±s)

组别	癌胚抗原(ng/ml)				癌抗原125(U/ml)				癌抗原199(U/ml)
组别	用药前	用药后	t值	P值	用药前	用药后	t值	P值	用药前	用药后	t值	P值
对照组(n＝16)	3.6±0.9	3.5±0.7	0.249	0.774	41.2±19.8	48.4±24.2	－0.782	0.294	23.6±11.7	31.3±28.0	－0.788	0.352
用药组(n＝12)	2.9±1.6	2.6±0.4^a	0.513	0.639	25.3±14.1	19.5±11.2^a	1.947	0.063	16.8±8.9	15.8±5.9	0.294	0.724
t值	1.683	2.856			1.982	2.645			1.167	1.711
P值	0.129	0.013			0.094	0.041			0.364	0.153

表4 两组患者用药前后肿瘤标志物比较(±s)

组别	癌胚抗原(ng/ml)				癌抗原125(U/ml)				癌抗原199(U/ml)
组别	用药前	用药后	t值	P值	用药前	用药后	t值	P值	用药前	用药后	t值	P值
对照组(n＝16)	3.6±0.9	3.5±0.7	0.249	0.774	41.2±19.8	48.4±24.2	－0.782	0.294	23.6±11.7	31.3±28.0	－0.788	0.352
用药组(n＝12)	2.9±1.6	2.6±0.4^a	0.513	0.639	25.3±14.1	19.5±11.2^a	1.947	0.063	16.8±8.9	15.8±5.9	0.294	0.724
t值	1.683	2.856			1.982	2.645			1.167	1.711
P值	0.129	0.013			0.094	0.041			0.364	0.153

表5 叶酸水平与miR-150相对含量、肿瘤标志物的相关性分析

皮尔逊相关系数	miR-150-5p	癌胚抗原(ng/ml)	癌抗原125(U/ml)	癌抗原199(U/ml)
r值	－0.621	－0.523	－0.308	－0.51
P值	0.025^a	0.081	0.331	0.932

表5 叶酸水平与miR-150相对含量、肿瘤标志物的相关性分析

皮尔逊相关系数	miR-150-5p	癌胚抗原(ng/ml)	癌抗原125(U/ml)	癌抗原199(U/ml)
r值	－0.621	－0.523	－0.308	－0.51
P值	0.025^a	0.081	0.331	0.932

图1 四款靶基因预测软件预测的miR-150-5p下游靶基因交集情况韦恩图注：Targetscan：靶基因预测软件http：//www.targetscan.org/预测的miR-150-5p下游靶基因；CLIP_Seq：锐博生物公司高通量CLIP-seq数据预测的miR-150-5p下游靶基因；miRDB：靶基因预测软件http：//mirdb.org/miRDB/预测的miR-150-5p下游靶基因；miRanda：靶基因预测软件http：//www.microrna.org/预测的miR-150-5p下游靶基因

图2 miR-150-5p下游靶基因京都基因与基因组百科全书通路富集程度分析注：SNARE：soluble N-ethylmaleimide-sensitive factor attachment protein receptor，可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体；FoxO：forkhead box O，叉头框蛋白O；ErbB：erythroblastic leukemia viral oncogene homolog，红母细胞白血病病毒同源性癌基因

[1]	Chen X, Li Y, Ding X, et al. Incidence, risk, and prognosis of cancer in patients on chronic hemodialysis [J]. Blood Purif, 2020, 49(3): 310-321.
[2]	Pieroth R, Paver S, Day S, et al. Folate and its impact on cancer risk [J]. Curr Nutr Rep, 2018, 7(3): 70-84.
[3]	Yuan S, Carter P, Vithayathil M, et al. Genetically predicted circulating B vitamins in relation to digestive system cancers [J]. Br J Cancer, 2021, 124(12): 1997-2003.
[4]	Kawakita D, Lee YA, Gren LH, et al. The impact of folate intake on the risk of head and neck cancer in the prostate, lung, colorectal, and ovarian cancer screening trial (PLCO) cohort [J]. Br J Cancer, 2018, 118(2): 299-306.
[5]	He Z, Dang J, Song A, et al. The involvement of miR-150/β-catenin axis in colorectal cancer progression [J]. Biomed Pharmacother, 2020, 121: 109495.
[6]	王珊，马清，杨华昱，等. 不同年龄段人群尿液外泌体中miR-150-5p对肾脏衰老的影响[J]. 中华老年医学杂志，2021, 40(11): 1386-1391.
[7]	Zhang DQ, Li HW, Chen HP, et al. Combination of amino-terminal pro-BNP, estimated GFR, and high-sensitivity CRP for predicting cardiorenal syndrome type 1 in acute myocardial infarction patients [J]. J Am Heart Assoc, 2018, 7(19): e009162.
[8]	王珊，杨华昱，姚兰，等. miR-21表达量与动脉粥样硬化程度的关系及长春西汀对其影响[J]. 中国医药导报，2017, 19(6)：975-977.
[9]	Shulpekova Y, Nechaev V, Kardasheva S, et al. The concept of folic acid in health and disease [J]. Molecules, 2021, 26(12): 3731.
[10]	Oliai Araghi S, Kiefte-de Jong JC, van Dijk SC, et al. Folic acid and vitamin B12 supplementation and the risk of cancer: long-term follow-up of the B vitamins for the prevention of osteoporotic fractures (B-PROOF) trial [J]. Cancer Epidemiol Biomarkers Prev, 2019, 28(2): 275-282.
[11]	Kanarek N, Petrova B, Sabatini DM. Dietary modifications for enhanced cancer therapy [J]. Nature, 2020, 579(7800): 507-517.
[12]	Senti FR, Pilch SM. Analysis of folate data from the second National Health and Nutrition Examination Survey (NHANES II) [J]. J Nutr, 1985, 115(11): 1398-1402.
[13]	Ivanovski N, Stojceva-Taneva O, Grozdanovski R, et al. Short-term effect of folic acid supplementation in renal transplant recipients and chronic kidney disease patients with comparable renal function impairment [J]. Nephrologie, 2004, 25(7): 301-303.
[14]	Mayne ST, Risch HA, Dubrow R, et al. Nutrient intake and risk of subtypes of esophageal and gastric cancer [J]. Cancer Epidemiol Biomarkers Prev, 2001, 10(10): 1055-1062.
[15]	Giovannucci E, Stampfer MJ, Colditz GA, et al. Multivitamin use, folate, and colon cancer in women in the Nurses′ Health Study [J]. Ann Intern Med, 1998, 129(7): 517-524.
[16]	Vollset SE, Clarke R, Lewington S, et al. Effects of folic acid supplementation on overall and site-specific cancer incidence during the randomised trials: meta-analyses of data on 50，000 individuals [J]. Lancet, 2013, 381(9871): 1029-1036.
[17]	Hill M, Tran N. miRNA interplay: mechanisms and consequences in cancer [J]. Dis Model Mech, 2021, 14(4): dmm047662.
[18]	Chen L, Heikkinen L, Wang C, et al. Trends in the development of miRNA bioinformatics tools [J]. Brief Bioinform, 2019, 20(5): 1836-1852.
[19]	Zhang J, Luo N, Luo Y, et al. microRNA-150 inhibits human CD133-positive liver cancer stem cells through negative regulation of the transcription factor c-Myb [J]. Int J Oncol, 2012, 40(3): 747-756.
[20]	Liu H, Liu R, Hao M, et al. Kinesin family member 3C (KIF3C) is a novel non-small cell lung cancer (NSCLC) oncogene whose expression is modulated by microRNA-150-5p (miR-150-5p) and microRNA-186-3p (miR-186-3p) [J]. Bioengineered, 2021, 12(1): 3077-3088.
[21]	Aherne ST, Madden SF, Hughes DJ, et al. Circulating miRNAs miR-34a and miR-150 associated with colorectal cancer progression [J]. BMC Cancer, 2015, 15: 329.
[22]	Zhang ZC, Wang GP, Yin LM, et al. Increasing miR-150 and lowering HMGA2 inhibit proliferation and cycle progression of colon cancer in SW480 cells [J]. Eur Rev Med Pharmacol Sci, 2018, 22(20): 6793-6800.

[1]	陈晓玲, 钟永洌, 刘巧梨, 李娜, 张志奇, 廖威明, 黄桂武. 超高龄髋膝关节术后谵妄及心血管并发症风险预测[J/OL]. 中华关节外科杂志(电子版), 2024, 18(05): 575-584.
[2]	陈翠萍, 李佩君, 杜景榕, 谢青梅, 许一宁, 卓姝妤, 李晓芳. 互联网联合上门护理在老年全髋关节置换术后的应用效果[J/OL]. 中华关节外科杂志(电子版), 2024, 18(05): 676-681.
[3]	宋俊锋, 张珍珍. 单侧初发性腹股沟斜疝老年患者经腹腹膜前疝修补术中残余疝囊腹直肌下缘固定效果评估[J/OL]. 中华疝和腹壁外科杂志(电子版), 2024, 18(06): 670-674.
[4]	张晋伟, 董永红, 王家璇. 基于GBD2021 数据库对中国与全球老年人疝疾病负担和健康不平等的分析比较[J/OL]. 中华疝和腹壁外科杂志(电子版), 2024, 18(06): 708-716.
[5]	邵世锋, 肖钦, 沈方龙, 张迅, 郝志鹏, 伍正彬, 谢晓娟, 王耀丽. 老年胸主动脉钝性伤的重症救治分析[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(05): 762-767.
[6]	赵月, 田坤, 张宗明, 郭震天, 刘立民, 张翀, 刘卓. 降钙素原对老年急性重度胆囊炎发生的预测价值[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 801-806.
[7]	张宗明, 董家鸿, 何小东, 王秋生, 徐智, 刘立民, 张翀. 老年胆道外科热点问题的争议与思考[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 754-762.
[8]	黄韬, 杨晓华, 薛天森, 肖睿. 改良“蛋壳”技术治疗老年OVCF及对脊柱矢状面平衡参数、预后的影响[J/OL]. 中华老年骨科与康复电子杂志, 2024, 10(06): 340-348.
[9]	王贝贝, 崔振义, 王静, 王晗妍, 吕红芝, 李秀婷. 老年股骨粗隆间骨折患者术后贫血预测模型的构建与验证[J/OL]. 中华老年骨科与康复电子杂志, 2024, 10(06): 355-362.
[10]	房桂彬, 肖进, 傅光涛, 郑秋坚. 老年髋部骨折患者术后1年行走能力的影响因素分析[J/OL]. 中华老年骨科与康复电子杂志, 2024, 10(05): 273-280.
[11]	王国强, 张纲, 唐建坡, 张玉国, 杨永江. LINC00839 调节miR-17-5p/WEE1 轴对结直肠癌细胞增殖、凋亡和迁移的影响[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(06): 491-499.
[12]	崔健, 夏青, 林云, 李光玲, 李心娜, 王位. 血小板与淋巴细胞比值、免疫球蛋白、心肌酶谱及心电图对中老年肝硬化患者病情及预后的影响[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(05): 400-406.
[13]	王誉英, 刘世伟, 王睿, 曾娅玲, 涂禧慧, 张蒲蓉. 老年乳腺癌新辅助治疗病理完全缓解的预测因素分析[J/OL]. 中华临床医师杂志(电子版), 2024, 18(07): 641-646.
[14]	周倩妹, 王宪娥, 徐筱, 老慧琳, 赵欣悦, 胡菁颖. 多元化系统护理对老年人群牙周健康指标影响的系统评价[J/OL]. 中华临床医师杂志(电子版), 2024, 18(05): 500-506.
[15]	赵伟伟, 赵玉华, 刘小璇. 西藏地区亚甲基四氢叶酸还原酶C677T多态性及其与脑微出血的相关性[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(05): 473-478.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Correlation between folic acid therapy and serum level of miR-150-5p in elderly patients undergoing maintenance hemodialysis

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Correlation between folic acid therapy and serum level of miR-150-5p in elderly patients undergoing maintenance hemodialysis