[1]岳芳芳a,倪文娟a,冷小敏b,等.人白介素10荧光定量PCR检测方法的建立及其在糖尿病肾病中的表达研究[J].现代检验医学杂志,2019,34(04):11-14.[doi:10.3969/j.issn.1671-7414.2019.04.003]
 YUE Fang-fanga,NI Wen-juana,LENG Xiao-minb,et al.Establishment of Real-Time Quantitative PCR Assay for Human IL-10 and Its Expression in Diabetic Nephropathy[J].Journal of Modern Laboratory Medicine,2019,34(04):11-14.[doi:10.3969/j.issn.1671-7414.2019.04.003]
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人白介素10荧光定量PCR检测方法的建立及其在糖尿病肾病中的表达研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第34卷
期数:
2019年04期
页码:
11-14
栏目:
论 著
出版日期:
2019-08-20

文章信息/Info

Title:
Establishment of Real-Time Quantitative PCR Assay for Human IL-10 and Its Expression in Diabetic Nephropathy
文章编号:
1671-7414(2019)04-011-04
作者:
岳芳芳a倪文娟a冷小敏b刘 云a师晶晶a曹东东a郭明好a马东红a
(新乡医学院第一附属医院a.肾脏病医院肾脏免疫研究所; b.生命科学研究中心,河南卫辉 453100)
Author(s):
YUE Fang-fangaNI Wen-juanaLENG Xiao-minbLIU YunaSHI Jing-jinga CAO Dong-dongaGUO Ming-haoaMA Dong-honga
(a.Institute of Nephrology and Immunology,the Hospital of Nephrology; b.Life Science Research Center, the First Affiliated Hospital of Xinxiang Medical University,Henan Weihui453100,China)
关键词:
白介素10 实时荧光定量PCR 糖尿病肾病
分类号:
Q503; R587.2
DOI:
10.3969/j.issn.1671-7414.2019.04.003
文献标志码:
A
摘要:
目的 建立一种稳定、灵敏的人白介素-10(IL-10)荧光定量PCR的检测方法,比较白介素10在糖尿病肾病组和对照组中的表达差异。方法取糖尿病肾病Ⅳ期患者的外周静脉血淋巴细胞RNA标本,主要从模板浓度、引物筛选两方面进行探索,建立稳定、灵敏的实验方法; 将建立的实验方法用于比较33例糖尿病肾病患者和35例正常健康组中血液白介素10的表达差异。结果 ①将475ng的总RNA逆转录成cDNA模板进行稀释,当模板浓度稀释范围在103倍以内,扩增引物为F:5'-GTGATGCCCCAAGCTGAGA-3',R:5'-CACGGCCTTGCTCTTGTTTT-3'时可以获得白介素10荧光定量PCR检测的最佳条件。②与对照组相比,糖尿病肾病组患者外周血白介素10的表达水平明显下调(糖尿病肾病组16.21±1.84 vs 对照组14.39±1.14),差异有统计学意义(t=7.049,P<0.01)。结论 该研究建立了检测白介素10的稳定的荧光定量PCR方法; 应用该检测方法发现,白介素10在糖尿病肾病组中的表达明显下降,为白介素10在糖尿病肾病中的功能研究和临床检测提供参考和借鉴。

参考文献/References:

[1] 杨颖涛.白细胞介素10基因多态性与食管癌的关系及其机制研究[D].郑州:郑州大学,2017. YANG Yingtao.The relationship between IL-10 ge-ne polymorphisms and esophageal cancer and its preliminary molecular mechanisms[D].Zhengzhou:Zhengzhou University,2017. [2] AL-SHUKAILI A,AL-GHAFRI S,AL-MARHOO-BI S,et al.Analysis of inflammatory mediators in type 2 diabetes patients[J].International Journal of Endocrinology,2013,2013:976810. [3] WONG C K,HO A W,TONG P C,et al.Aberrant activation profile of cytokines and mitogen-activated protein kinases in type 2 diabetic patients with nephropathy[J].Clin Exp Immunol,2007,149(1):123-131. [4] YUAN Ning,ZHANG Haifeng,WEI Qi,et al.Expression of CD4+CD25+Foxp3+ regulatory T cells,interleukin 10 and transforming growth factor β in newly diagnosed type 2 diabetic patients[J].Exp Clin Endocrinol Diabetes,2018,126(2):96-101. [5] YAGHINI N,MAHMOODI M,ASADIKARAM G R,et al.Serum levels of interleukin10(IL-10)in patients with type 2 diabetes[J].Iran Red Crescent Med J,2011,13(10):752. [6] 中华医学会糖尿病学分会.中国2型糖尿病防治指南(2017年版)[J].中国实用内科杂志,2018,38(4):292-344. Chinese Diabetes Society.Guidelines for the prevention and controlof type 2 diabetes in China(2017 Edition)[J].Chinese Journal of Practical Internal Medicine,2018,38(4):292-344. [7] WADA J,MAKINO H.Inflammation and the pathogenesis of diabetic nephropathy[J].Clin Sci,2013,124(3):139-152. [8] 陈显,朱绍汶,黄成垠,等.ELISA法检测抗-HCV灰区设置探讨[J].临床输血与检验,2013,15(3):243-244. CHEN Xian,ZHU Shaowen,HUANG Chengyin,et al.Discussion on the detection of anti-HCV gray zone by ELISA[J].Journal of Clinical Transfusion andLaboratory Medicine,2013,15(3):243-244. [9] ORRUˋ G,OCCHINEGRO A,RATTO D,et al.MiRNA-155 and colorectal cancer,the role of real time PCR in laboratory diagnosis[J].Eur Rev Med Pharmacol Sci,2018,22(6):1521-1524. [10] PENG Juan,XIE Zhengyong,CHENG Liyang,et al.Paired design study by real-time PCR:miR-378* and miR-145 are potent early diagnostic biomarkers of human colorectal cancer[J].BMC Cancer,2015,15(1):158. [11] 曹锡梅,梁俊红,张潮,等.均匀设计在候选内参基因RT-qPCR实验条件优化中的应用[J].中国组织化学与细胞化学杂志,2012,21(6):522-527. CAO Ximei,LIANG Junhong,ZHANG Chao,et al.Application of uniform designto optimize quantitative real-time RT-qPCR conditions in potential internal control genes[J].Chinese Journal of Histochemistry and Cytochemistry,2012,21(6):522-527.

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备注/Memo

备注/Memo:
基金项目:河南省科技厅重点研发与推广项目(182102310585); 河南省高等学校重点科研项目(17A320026); 河南省卫生厅科技攻关省部共建备选项目(2018010013)。 作者简介:岳芳芳(1991-),女,硕士,住院医师,研究方向:糖尿病肾病,E-mail:1010208531@qq.com。 通讯作者:郭明好(1964-),男,硕士,教授,硕士研究生导师,研究方向:肾小球疾病,E-mail:guomh@163.com。 马东红(1980-),女,博士,副教授,硕士研究生导师,研究方向:糖尿病肾病,E-mail:dhmasxmu@163.com。收稿日期:2019-03-14 修回日期:2019-04-23
更新日期/Last Update: 2019-07-30