Binding characteristics of chemosynthetic Ac-SDKP analogue FAM-Aca-SDKP to hepatic stellate cell-T6 cells
10.3760/cma.j.issn.1007-3418.2016.03.006
- VernacularTitle:化学合成的Ac-SDKP类似物FAM-Aca-SDKP与HSC-T6细胞表面结合特征分析
- Author:
Da ZHOU
1
;
Lingnan HE
;
Jing WANG
;
Yongnian DING
;
Yuanwen CHEN
;
Jiangao FAN
Author Information
1. 200092,上海交通大学医学院附属新华医院消化内科
- Keywords:
Liver cirrhosis;
Hepatic stellate cell;
Receptor;
Ligand;
Ac-SDKP;
Binding characteristics
- From:
Chinese Journal of Hepatology
2016;24(3):186-190
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the binding of the chemosynthetic analogue of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) FAM-Aca-SDKP to hepatic stellate cell-T6 (HSC-T6) cells and basic physical characteristics.Methods The Ac-SDKP analogue short-peptide FAM-Aca-SDKP carrying green fluorescence was synthesized chemically.Quantitative real-time PCR was used to evaluate its effect on the secretion of HSC collagen and verify the consistency in the biological effect between FAM-Aca-SDKP and Ac-SDKP.A fluorescence microscope was used to observe the binding between FAM-Aca-SDKP and HSC-T6,and flow cytometry was used to evaluate the time-concentration effect of the binding between FAM-Aca-SDKP and HSC-T6.The t-test or rank sum test was used for the statistical analysis of different types ofdata.Results After HSC-T6 was incubated with Ac-SDKP or FAM-Aca-SDKP for 24 hours,the expression of type Ⅰ collagen in HSC-T6 was increased,when the action time was 0.5 hour,Ac-SDKP and FAM-Aca-SDKP caused a 30％-50％ reduction in the expression of type Ⅰ collagen.After HSC-T6 was incubated with FAM-Aca-SDKP,strong green fluorescence was observed on cell surface under a fluorescence microscope,and after Ac-SDKP was added,Ac-SDKP significantly reduced the fluorescence intensity on cell surface due to competitive inhibition.Flow cytometry showed that when the concentration of FAM-Aca-SDKP was 0-50 μmol/L,the rate of fluorescence-positive cells rapidly increased from 0 to 12％;when the concentration was 50-100 μmol/L,the rate of fluorescence-positive cells only increased from 12％ to 14％;co-incubation with Ac-SDKP significantly reduced the rate of fluorescence-positive cells.The number of positive cells reached the peak at the 45-minute point of the incubation and then decreased gradually.Conclusions FAM-Aca-SDKP can bind to the surface of HSC-T6 cells,and this process has ligand-receptor binding characteristics such as competitive inhibition,saturability,and time-concentration effect.
