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

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[15]	晏美娟, 邵礼晖. 高水平脂蛋白（a）与无“三高”老年人群小动脉硬化型脑小血管病的相关性研究[J]. 中华脑血管病杂志(电子版), 2023, 17(05): 458-463.

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

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