Expression and secretion of TGF-beta2 in human retinal pigment epithelium cell line D407 regulated by atropine.
10.3969/j.issn.1672-7347.2010.05.018
- Author:
Jia TAN
1
;
Zhihong DENG
;
Shuangzhen LIU
Author Information
1. Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha 410008, China.
- Publication Type:Journal Article
- MeSH:
Adult;
Atropine;
pharmacology;
Carbachol;
pharmacology;
Cell Line;
Female;
Humans;
Male;
Muscarinic Antagonists;
pharmacology;
RNA, Messenger;
genetics;
metabolism;
Retinal Pigment Epithelium;
cytology;
metabolism;
Transforming Growth Factor beta2;
genetics;
metabolism;
Young Adult
- From:
Journal of Central South University(Medical Sciences)
2010;35(5):518-523
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the regulation of atropine to the expression and secretion of TGF-beta2 in retinal pigment epithelium (RPE) cells by observing the changes of those under different treatments of atropine and carbachol.
METHODS:D407 cells were cultured conventionally and divided into 4 groups as follows: (1) An experimental group (Group A), cells were pretreated with 10(-4)-10(-8) mol/L atropine for 30 min, and then treated with 10(-5) mol/L carbachol; (2) a negative control group (Group B), cells were treated with 10(-4)-10(-8) mol/L atropine; (3) a positive control group (Group C), cells were treated with 10(-5) mol/L carbachol; (4) a blank control group (Group D). The concentration of TGF-beta2 in the supernate, and the level of TGF-beta2 mRNA and protein were measured by ELISA, RT-PCR, and Western blot after the 24-hour treatment. The data were analyzed by analysis of variance.
RESULTS:The levels of TGF-beta2 mRNA and protein in the cytoplasm and the concentration of TGF-beta2 in the supernate in the experimental groups were lower than those of the positive control group. Atropine at 10-4 mol/L could completely inhibit the effect of carbachol at 10-5 mol/L. The effect of atropine was concentration-dependent (F=1,056.897,1,320.170, and 475.657; P<0.001). There was no change of TGF-beta2 level in the cytoplasm and supernate with the treatment of atropine alone (P>0.05).
CONCLUSION:Carbachol can promote the expression and secretion of TGF-beta2 in human RPE cells and atropine could reverse it effectively, suggesting that M receptor may be involved.
