期刊论文

Validation of effect of IMM-H004 on CKLF1/CCR4 combination based on BiFC

中文

中国药理学通报

1001-1978

2020

36

7

1030-1035

10.3969/j.issn.1001-1978.2020.07.025

No81873026:国家自然科学基金 81730096:国家自然科学基金 No2018ZX09711001-003-005:国家科技重大专项 2018ZX09711001-009-013:国家科技重大专项 No2016-I2M-1-004:创新工程项目

本校为第一机构

目的利用BiFC双分子荧光互补技术研究香豆素衍生物IMM-H004对趋化素样因子CKLF1与其受体CCR4结合的影响。方法引入BiFC技术,将CKLF1趋化活性肽段C27与CCR4分别与Venus荧光蛋白的N端(VN173)和C端(VC155)相连,C27与CCR4的结合可以Venus黄色荧光的形式体现,重组载体共转染HEK293细胞,进行氧糖剥夺/复氧损伤造模,并给予0.1、1和10 μmol·L~(-1) IMM-H004,观察活细胞中荧光强度。结果通过测序比对验证CKLF1/CCR4-BiFC系统构建成功,并能转染HEK293细胞正常启动,在该系统中,与Control组相比,OGD/R造模后黄色荧光强度明显增加(P < 0.01),给予不同浓度IMM-H004后荧光表达均明显降低(P < 0.01),差异具有统...

Aim To study the effect of coumarin derivative IMMH004 on the binding of chemokine-like factor (CKLFI) to its receptor CCR4 using bimolecular fluorescence complementary technology. Methods The BiFC technology was introduced to this research. The CKLFl chemotactic peptides C27 and CCR4 were connected to the N-terminus (VN173) and C-terminus (VC155) of the Venus fluorescent protein. The combination of C27 and CCR4 can be reflected in the form of Venus yellow fluorescence. HEK293 cells were transfected, and oxygen-glucose deprivation/reoxygenation...