[1]吴 雷,叶树培,黄玉娥,等.过表达受体活性修饰蛋白-1对人气道平滑肌细胞增殖和细胞周期的影响[J].郑州大学学报(医学版),2019,(04):579-582.[doi:10.13705/j.issn.1671-6825.2018.10.118]
　WU Lei,YE Shupei,HUANG Yu'e,et al.Effects of receptor activity-modifying protein-1 overexpression on proliferation and cell cycle of human airway smooth muscle cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2019,(04):579-582.[doi:10.13705/j.issn.1671-6825.2018.10.118]
点击复制

过表达受体活性修饰蛋白-1对人气道平滑肌细胞增殖和细胞周期的影响()
分享到：

《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2019年04期
页码:
579-582
栏目:
应用研究
出版日期:
2019-07-20

文章信息/Info

Title:
Effects of receptor activity-modifying protein-1 overexpression on proliferation and cell cycle of human airway smooth muscle cells
作者:
吴 雷; 叶树培; 黄玉娥; 陈翠仪; 陈美华
东莞市第三人民医院呼吸内科 广东东莞 523326
Author(s):
WU Lei; YE Shupei; HUANG Yu'e; CHEN Cuiyi; CHEN Meihua
Department of Respiratory Medicine, the Third People's Hospital of Dongguan,Dongguan,Guangdong 523326
关键词:
气道平滑肌细胞;  受体活性修饰蛋白-1;  NF-κB信号通路
Keywords:
airway smooth muscle cell;  receptor activity-modifying protein-1;  NF-κB signaling pathway
分类号:
R562.2
DOI:
10.13705/j.issn.1671-6825.2018.10.118
摘要:
目的:探讨过表达受体活性修饰蛋白-1(RAMP)对人气道平滑肌细胞(ASMCs)增殖和细胞周期的影响及机制。方法:从人正常肺叶支气管分离出ASMCs,取第3～8代细胞,转染重组pcDNA3.1-RAMP-1,同时将转染pcDNA3.1空载体的细胞作对照,并设置空白组。转染48 h后,Western blot检测RAMP-1、IκBα、p65NF-κB、PCNA和CyclinD1蛋白表达,流式细胞术检测细胞周期。CCK-8法检测转染24、48和72 h后细胞增殖情况。结果:pcDNA3.1-RAMP-1组ASMCs中RAMP-1蛋白表达明显高于空白组(P<0.05)。与空白组和pcDNA3.1组比较,pcDNA3.1-RAMP-1组细胞增殖降低,G0/G1期细胞比例升高,而S期和G2/M期细胞比例降低,IκBα蛋白表达升高,p65NF-κB、PCNA和CyclinD1蛋白表达降低(P<0.05)。结论:RAMP-1可抑制人ASMCs增殖,阻滞细胞周期进程,机制可能与抑制NF-κB信号通路有关。
Abstract:
Aim:To investigate the effects of overexpressed receptor activity-modifying protein-1(RAMP-1)on proliferation and cell cycle of human airway smooth muscle cells(ASMCs)and its mechanism.Methods:ASMCs were isolated from human normal pulmonary lobe bronchus and cultured.Recombinant pcDNA3.1-RAMP-1 was transfected into ASMCs, and ASMCs transfected with pcDNA3.1 empty vector was used as control group, and a blank group was set up. After cells were transfected for 48 hours, the expressions of RAMP-1, IκBα, p65NF-κB, PCNA and CyclinD1 protein were detected by Western blot,and the cell cycle was detected by flow cytometry. CCK-8 assay was used to detect cell proliferation after cells were transfected for 24, 48 and 72 hours.Results:The expression of RAMP-1 in ASMCs transfected with recombinant pcDNA3.1-RAMP-1 was significantly higher than that in blank group(P<0.05). Compared with the blank and pcDNA3.1 groups, the cell proliferation in pcDNA3.1-RAMP-1 group was decreased, the proportion of G0/G1 phase cells was increased, the proportion of S phase and G2/M phase cells was decreased, the expression of IκBα was increased, and the expression of p65NF-κB, PCNA and CyclinD1 were decreased(P<0.05).Conclusion:RAMP-1 can inhibit the proliferation and cell cycle progression of ASMCs, and the mechanism may be related to the inhibition of NF-κB signaling pathway.

参考文献/References:

[1] RAHMAN MA,CHAKRABORTY R,FERDOUSI KR,et al.New therapeutic approach to treat allergic rhinitis&bronchial asthma,considering these two as one united airway disease[J].Mymensingh Med J,2017,26(1):216
[2] NAKAMURA Y,SUZUKI R,MIZUNO T,et al.Therapeutic implication of genetic variants of IL13 and STAT4 in airway remodelling with bronchial asthma[J].Clin Exp Allergy,2016,46(9):1152
[3] WEN X,YAN J,HAN XR,et al.PTEN gene silencing contributes to airway remodeling and induces airway smooth muscle cell proliferation in mice with allergic asthma[J].J Thorac Dis,2018,10(1):202
[4] 郑秀丽,杨宇,王宝家,等.过敏性哮喘大鼠模型结肠和肺组织CCK-8、CGRP、SP及VIP变化观察[J].中国中西医结合杂志,2013,33(12):1668
[5] JUSEK G,REIM D,TSUJIKAWA K,et al.Deficiency of the CGRP receptor component RAMP1 attenuates immunosuppression during the early phase of septic peritonitis[J].Immunobiology,2012,217(8):761
[6] MARTIN VT,FANNING KM,SERRANO DA,et al.Asthma is a risk factor for new onset chronic migraine: results from the American migraine prevalence and prevention study[J].Headache,2016,56(1):118
[7] KIM SH,PEI QM,JIANG P,et al.Role of licochalcone A in VEGF-induced proliferation of human airway smooth muscle cells: implications for asthma[J].Growth Factors,2017,35(1):39
[8] 张晓宇,马利军,郭亚丽,等.BCYRN1对大鼠哮喘模型中气管平滑肌细胞增殖和迁移的影响[J].中华医学杂志,2016,96(46):3751
[9] EDVINSSON L.The trigeminovascular pathway:role of CGRP and CGRP receptors in migraine[J].Headache,2017,57(Suppl 2):47
[10]ZHANG Y,XU J,RUAN YC,et al.Implant-derived magnesium induces local neuronal production of CGRP to improve bone-fracture healing in rats[J].Nat Med,2016,22(10):1160
[11]HAY DL,WALKER CS.CGRP and its receptors[J].Headache,2017,57(4):625
[12]郑元斌,秦旭平,孙飞,等.受体活性修饰蛋白1基因载体构建及其对大鼠血管平滑肌细胞增殖的影响[J].南华大学学报(医学版),2010,38(1):1
[13]石蓓,崔璨,龙仙萍,等.CGRP修饰的间充质干细胞及其CGRP受体对血管平滑肌细胞增殖及表型转化的影响[J].中华医学杂志,2013,93(30):2372
[14]CILDIR G,LOW KC,TERGAONKAR V.Noncanonical NF-κB signaling in health and disease[J].Trends Mol Med,2016,22(5):414
[15]SAU A,LAU R,CABRITA MA,et al.Persistent activation of NF-κB in BRCA1-deficient mammary progenitors drives aberrant proliferation and accumulation of DNA damage[J].Cell Stem Cell,2016,19(1):52
[16]LEE IS,UH IJ,KIM KS,et al.Anti-inflammatory effects of ginsenoside Rg3 via NF-κB pathway in A549 cells and human asthmatic lung tissue[J].J Immunol Res,2016,2016:7521601
[17]石惠玲,刘杰波,卢爱萍.PI3K、NF-κB和STAT1在支气管哮喘儿童外周血单个核细胞的表达和临床意义[J].中国当代儿科杂志,2016,18(7):614
[18]龙仙萍,崔璨,陈攀科,等.降钙素基因相关肽和受体活性修饰蛋白1对血管平滑肌细胞迁移的影响及机制[J].中华心血管病杂志,2015,43(6):537
[19]WANG CY,LI HT,CAO LF,et al.Identification of differentially expressed genes associated with asthma in children based on the bioanalysis of the regulatory network[J].Mol Med Rep,2018,18(2):2153
[20]DU CL,XU YJ,LIU XS,et al.Up-regulation of cyclin D1 expression in asthma serum-sensitized human airway smooth muscle promotes proliferation via protein kinase C alpha[J].Exp Lung Res,2010,36(4):201

备注/Memo

备注/Memo:
【基金项目】东莞市科技计划项目(201210515024072)
【作者简介】陈美华,通信作者,女,1964年6月生,本科,主任医师,研究方向:慢性气道疾病,E-mail:fanxi98849988@163.com
更新日期/Last Update: 2019-07-20