特应性皮炎患者血浆外泌体miR-21和lncRNA MALAT1的检测水平及意义

蔡宝祥; 朱健伟; 陈艳; 金瀛凯

doi:10.16766/j.cnki.issn.1674-4152.002363

特应性皮炎患者血浆外泌体miR-21和lncRNA MALAT1的检测水平及意义

doi: 10.16766/j.cnki.issn.1674-4152.002363

1.
宁波市医疗中心李惠利医院皮肤科，浙江宁波 315040
2.
浙江医院皮肤科，浙江杭州 310013

基金项目:

浙江省基础公益研究计划项目 LY20H070006

详细信息

通讯作者:
蔡宝祥，E-mail：baoxiangcai9177@163.com

中图分类号: R758.29
计量
- 文章访问数: 143
- HTML全文浏览量: 107
- PDF下载量: 4
- 被引次数: 0
出版历程
- 收稿日期: 2021-05-20
- 网络出版日期: 2022-08-13

Detection and significance of plasma exosomes miR-21 and lncRNA MALAT1 in patients with atopic dermatitis

1.
Department of Dermatology, Ningbo Medical Center Li Huili Hospital, Ningbo, Zhejiang 315040, China

摘要

摘要: 目的分析血浆外泌体微小RNA-21(miR-21)、长链非编码RNA(lncRNA)肺癌转移相关转录本1(MALAT1)在特应性皮炎(AD)患者中的表达水平，探讨二者的临床意义。方法选择2018年1月—2020年5月宁波市医疗中心李惠利医院皮肤科收治的AD患者118例，依据特应性皮炎积分指数(SCORAD)将患者分为轻度41例、中度38例、重度39例，另选取同期健康体检者120例为对照组。提取并鉴定血浆外泌体；采用实时荧光定量PCR法(qRT-PCR)测定血浆外泌体中miR-21、lncRNA MALAT1水平；Pearson法分析血浆外泌体miR-21、lncRNA MALAT1水平与SCORAD评分的相关性；多元线性逐步回归模型分析AD患者SCORAD评分增加的危险因素。结果所提取样品具备外泌体特征；AD组轻、中、重度患者血浆外泌体中miR-21、lncRNA MALAT1水平、SCORAD评分显著高于对照组，且随病情严重程度增加而升高(F=91.160、206.502、246.615，均P＜0.001)。血浆外泌体miR-21、lncRNA MALAT1水平与SCORAD评分均呈正相关关系(r=0.631、0.528，均P＜0.05)。血浆外泌体miR-21、lncRNA MALAT1水平高均为AD患者SCORAD评分增加的危险因素(均P＜0.05)。结论血浆外泌体miR-21、lncRNA MALAT1异常高表达可能与AD发生发展有关，且与病情严重程度关系密切，监测二者水平变化可能对进一步靶向防治AD发生有一定的意义。
- 特应性皮炎 /
- 血浆外泌体 /
- 微小RNA-21 /
- 肺癌转移相关转录本1
Abstract: Objective To analyse the expression levels of plasma exosome microRNA-21 (miR-21) and long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in patients with atopic dermatitis (AD) and to explore their clinical significance. Methods A total of 118 patients with AD in the Dermatology Department of Ningbo Medical Center Li Huili Hospital from January 2018 to May 2020 were selected and divided into 41 cases of mild, 38 cases of moderate and 39 cases of severe according to the scoring atopic dermatitis index (SCORAD). At the same time, 120 healthy people were selected as the control group. Plasma exosomes were extracted and identified. The levels of miR-21 and lncRNA MALAT1 in plasma exosomes were measured by real-time fluorescence quantitative polymerase chain reaction. The Pearson method was used to analyse the correlation among miR-21, lncRNA MALAT1 and SCORAD score, and multiple linear stepwise regression model was used to analyse the risk factors of increased SCORAD score in patients with AD. Results The extracted samples had the characteristics of exosomes. The levels of serum miR-21, lncRNA MALAT1 and SCORAD score in the mild, moderate and severe patients in the AD group were significantly higher than those in the control group and increased with the increase in disease severity (F=91.160, 206.502, 246.615, all P < 0.001). The levels of miR-21 and lncRNA MALAT1 in plasma exosomes were positively correlated with SCORAD score (r=0.631, 0.528, all P < 0.05). The high levels of plasma exosomes miR-21 and lncRNA MALAT1 were the risk factors for the increase in SCORAD score in patients with AD (all P < 0.05). Conclusion The abnormally high expression levels of plasma exosomes miR-21 and lncRNA MALAT1 may be related to the occurrence and development of AD and closely related to disease severity. Monitoring the levels of miR-21 and lncRNA MALAT1 may be important for further targeted prevention and treatment of AD.
- Atopic dermatitis /
- Plasma exosomes /
- MicroRNA-21 /
- Metastasis-associated lung adenocarcinoma transcript 1

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图 1 Nano Sight检测结果

Figure 1. Nano Sight test results

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图 2 透射电镜观察结果

Figure 2. Transmission electron microscope observation results

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图 3 Western blotting检测结果

注：1为血细胞，2为外泌体样品，3为血浆。

Figure 3. Test results of Western blotting

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图 4 AD患者血浆外泌体miR-21水平与SCORAD评分的相关性分析

Figure 4. Multiple linear stepwise regression model analysisof SCORAD score in AD patients

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图 5 AD患者血浆外泌体lncRNA MALAT1水平与SCORAD评分的相关性分析

Figure 5. Correlation Analysis between plasma secreted lncRNAMALAT1 level and SCORAD score in AD patients

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表 1 miR-21、lncRNA MALAT1、U6引物序列

Table 1. Primer sequences of miR-21, lncRNA MALAT1 and U6

基因	正向引物5'-3'	反向引物5'-3'
miR-21	TGACAAAGGCAGGAGGTA	ATCTCTGGGTGCTGGTGAAGG
lncRNA MALAT1	TGCGGCAAAGTGCTTACAGTG	CCAGTGCAGGGTCCGAGGT
U6	AAGGTGAAGGTCGGAGTCAAC	GGGGTCATTGATGGCAACAATA

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表 2 各组研究对象血浆外泌体miR-21、lncRNA MALAT1水平及SCORAD评分比较(x±s)

Table 2. Comparison of plasma exosome miR-21, lncRNA MALAT1levels and SCORAD scores of subjects in each group (x±s)

组别	例数	miR-21	lncRNA MALAT1	SCORAD评分(分)
对照组	120	1.09±0.09	1.03±0.08
轻度AD组	41	1.86±0.68^a	1.97±0.50^a	11.82±3.20
中度AD组	38	2.33±0.96^ab	2.76±0.66^ab	29.69±8.77^b
重度AD组	39	2.79±1.05^abc	3.82±1.38^abc	71.87±19.38^bc
F值		91.160	206.502	246.615
P值		＜0.001	＜0.001	＜0.001
注：与对照组比较，^aP＜0.05；与轻度AD组比较，^bP＜0.05；与中度AD组比较，^cP＜0.05。

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表 3 变量赋值方法

Table 3. Variable assignment method

变量	赋值方法
miR-21	>2.32=1，＜2.32=0
lncRNA MALAT1	>2.84=1，＜2.84=0
注：以AD组118例患者血浆外泌体miR-21、lncRNA MALAT1平均值为自变量赋值标准。

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表 4 AD患者SCORAD评分的多元线性逐步回归模型分析

Table 4. Multiple linear stepwise regression model analysisof SCORAD score in AD patients

变量	B	SE	B'	t值	P值
miR-21	-5.441	2.183	-0.173	-2.492	0.014
lncRNA MALAT1	1.613	0.379	0.307	4.254	＜0.001

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参考文献(20)

[1]	FRAZIER W, BHARDWAJ N. Atopic dermatitis: Diagnosis and treatment[J]. Am Fam Physician, 2020, 101(10): 590-598.
[2]	PENG G, MU Z Z, CUI L X, et al. Anti-IL-33 antibody has a therapeutic effect in an atopic dermatitis murine model induced by 2, 4-dinitrochlorobenzene[J]. Inflammation, 2018, 41(1): 154-163. doi: 10.1007/s10753-017-0673-7
[3]	CHEN L, GUO P, HE Y C, et al. HCC-derived exosomes elicit HCC progression and recurrence by epithelial-mesenchymal transition through MAPK/ERK signalling pathway[J]. Cell Death Dis, 2018, 9(5): 513-516. doi: 10.1038/s41419-018-0534-9
[4]	蓝涛, 郑剑, 刘威, 等. 基于向量机算法的职业性三氯乙烯药疹样皮炎诊断模型的建立[J]. 华南预防医学, 2017, 43(5): 438-441. https://www.cnki.com.cn/Article/CJFDTOTAL-GDWF201705009.htm LAN T, ZHENG J, LIU W, et al. Establishment of diagnostic model for occupational medicamentosa-like dermatitis induced by trichloroethylene based on support vector machines[J]. South China Journal of Preventive Medicine, 2017, 43(5): 438-441. https://www.cnki.com.cn/Article/CJFDTOTAL-GDWF201705009.htm
[5]	SALLAM T, SANDHU J, TONTONOZ P. Long noncoding RNA discovery in cardiovascular disease: Decoding form to function[J]. Circ Res, 2018, 122(1): 155-166. doi: 10.1161/CIRCRESAHA.117.311802
[6]	PAN F, ZHU L, LV H, et al. Quercetin promotes the apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by upregulating lncRNA MALAT1[J]. Int J Mol Med, 2016, 38(5): 1507-1514. doi: 10.3892/ijmm.2016.2755
[7]	GAO F, TAN Y, LUO H. MALAT1 is involved in type Ⅰ IFNs-mediated systemic lupus erythematosus by up-regulating OAS2, OAS3, and OASL[J]. Braz J Med Biol Res, 2020, 53(5): e9292. doi: 10.1590/1414-431x20209292
[8]	慕彰磊, 张建中. 特应性皮炎的诊断标准[J]. 中国医学文摘(皮肤科学), 2016, 33(2): 97-100. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYXW201602003.htm MU Z L, ZHANG J Z. Diagnostic criteria for atopic dermatitis[J]. China Medical Abstracts of Dermatology, 2016, 33(2): 97-100. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYXW201602003.htm
[9]	ROSSI A B, BACQUEY A, NOCERA T, et al. Efficacy and tolerability of a medical device repairing emollient cream associated with a topical corticosteroid in adults with atopic dermatitis: An open-label, intra-individual randomized controlled study[J]. Dermatol Ther, 2018, 8(2): 217-228. doi: 10.1007/s13555-018-0228-3
[10]	王海燕, 陈达灿, 晏烽根. 特应性皮炎中西医发病机制的比较与思考[J]. 中国中西医结合杂志, 2018, 38(6): 737-740. https://www.cnki.com.cn/Article/CJFDTOTAL-ZZXJ201806023.htm WANG H Y, CHEN D C, YAN F G. Comparison and thinking on the pathogenesis of atopic dermatitis between traditional Chinese medicine and western medicine[J]. Chinese Journal of Integrated Traditional and Western Medicine, 2018, 38(6): 737-740. https://www.cnki.com.cn/Article/CJFDTOTAL-ZZXJ201806023.htm
[11]	CHOI C W, YANG B R, SUH D I, et al. Infection, antibiotic exposure and development of atopic dermatitis: A nationwide case-control study[J]. J Dermatol, 2020, 47(7): 707-713. doi: 10.1111/1346-8138.15387
[12]	ZHANG Y, WANG W T, GONG C R, et al. Combination of olfactory ensheathing cells and human umbilical cord mesenchymal stem cell-derived exosomes promotes sciatic nerve regeneration[J]. Neural Regen Res, 2020, 15(10): 1903-1911. doi: 10.4103/1673-5374.280330
[13]	罗靖莹, 贺宏丽, 郭阳, 等. 差速离心、密度梯度离心、超滤离心技术在骨髓间充质干细胞外泌体提取中的应用对比观察[J]. 山东医药, 2019, 59(12): 48-52. doi: 10.3969/j.issn.1002-266X.2019.12.013 LUO J Y, HE H L, GUO Y, et al. Comparison of differential centrifugation, density gradient centrifugation, and ultrafiltration of centrifugation in extraction of exosomes from bone marrow mesenchymal stem cells[J]. Shandong Medical Journal, 2019, 59(12): 48-52. doi: 10.3969/j.issn.1002-266X.2019.12.013
[14]	ZHANG Q L, DONG Z X, LUO Z W, et al. MicroRNA profile of immune response in gills of zebrafish (Danio rerio) upon Staphylococcus aureus infection[J]. Fish Shellfish Immunol, 2019, 87(4): 307-314.
[15]	CHEN N, FENG L, QU H, et al. Overexpression of IL-9 induced by STAT3 phosphorylation is mediated by miR-155 and miR-21 in chronic lymphocytic leukemia[J]. Oncol Rep, 2018, 39(6): 3064-3072. https://www.spandidos-publications.com/or/39/6/3064
[16]	MURUGAIYAN G, CUNHA A P, AJAY A K, et al. MicroRNA-21 promotes Th17differentiation and mediates experimental autoimmune encephalomyelitis[J]. J Clin Invest, 2015, 125(3): 1069-1080. doi: 10.1172/JCI74347
[17]	ZHU W, MEN X L. Negative feedback of NF-κB signaling by long noncoding RNA MALAT1 controls lipopolysaccharide-induced inflammation injury in human lung fibroblasts WI-38[J]. J Cell Biochem, 2020, 121(2): 1945-1952. doi: 10.1002/jcb.29429
[18]	SILVERBERG J I, LEI D, YOUSAF M, et al. Comparison of Patient-Oriented Eczema Measure and Patient-Oriented Scoring Atopic Dermatitis vs Eczema Area and Severity Index and other measures of atopic dermatitis: A validation study[J]. Ann Allergy Asthma Immunol, 2020, 125(1): 78-83. doi: 10.1016/j.anai.2020.03.006
[19]	林银哲, 黄咏菁, 莫秀梅, 等. 特应性皮炎患者中医证型与临床评价指标的关系研究[J]. 中国现代医学杂志, 2019, 29(13): 119-122. doi: 10.3969/j.issn.1005-8982.2019.13.023 LIN Y Z, HUANG Y J, MO X M, et al. Relationship between TCM syndromes and clinical evaluation indexes in patients with atopic dermatitis[J]. China Journal of Modern Medicine, 2019, 29(13): 119-122. doi: 10.3969/j.issn.1005-8982.2019.13.023
[20]	张绍夫, 王飞, 李航, 等. 急性缺血性脑卒中患者溶栓后血浆miR-146a、miR-21表达变化及意义[J]. 山东医药, 2019, 59(1): 61-63. doi: 10.3969/j.issn.1002-266X.2019.01.018 ZHANG S F, WANG F, LI H, et al Changes and significance of plasma miR-146a and miR-21 expression in patients with acute ischemic stroke after thrombolysis[J]. Shandong Medical Journal, 2019, 59(1): 61-63. doi: 10.3969/j.issn.1002-266X.2019.01.018