[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-fanga; NI Wen-juana; LENG Xiao-minb; LIU Yuna; SHI Jing-jinga;  CAO Dong-donga; GUO Ming-haoa; MA 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。
Abstract:
Objective A stable and sensitive method for thedetection of human interleukin-10(IL-10)real-time fluorescence quantitative PCR was established to compare the expression differences of IL-10 in diabetic nephropathy group and control group.Methods The peripheral venous blood lymphocyte RNA samples were obtained from patients with stage IV diabetic nephropathy.The expression level of IL-10 was detected by real-time quantitative PCR.This experiment mainly explored both template concentration and primer screening to establish a stable and sensitive experimental method for comparing the expression levels of IL-10 in 33 patients with diabetic nephropathy and 35 normal healthy groups.Results ①After reversing transcription of 475 ng of total RNA into cDNA,diluting the template,and when the template concentration dilution range was within 103 times,and the amplification primers were F:5'-GTGATGCCCCAAGCTGAGA-3',R:5'-CACGGCCTTGCTCTTGT TTT-3',it could obtain the optimal conditions for the detection of IL-10 fluorescence quantitative PCR.②Compared with the control group,the expression level of IL-10 in peripheral blood of patients with diabetic nephropathywas significantly down-regulated(diabetic nephropathy group 16.21±1.84 vscontrol group 14.39±1.14),and the difference was statistically significant(t=7.049,P<0.01).Conclusion The study established a stable real-time quantitative PCR method for the detection of IL-10.By this method,it was found that the expression of IL-10 was significantly decreased in the diabetic nephropathy group.It will provide reference for the functional research and clinical detection of IL-10 in diabetic nephropathy.

参考文献/References:

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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