[Expression and secretion of TGF-beta2 in human retinal pigment epithelium cell line D407 regulated by atropine].
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).


CONCLUSIONS
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.
Tan
Jia
J
Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha 410008, China.
Deng
Zhihong
Z
Liu
Shuangzhen
S
D000328:Adult; D001285:Atropine / Q000494:pharmacology*; D002217:Carbachol / Q000494:pharmacology*; D002460:Cell Line; D005260:Female; D006801:Humans; D008297:Male; D018727:Muscarinic Antagonists / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000378:metabolism*; D053781:Transforming Growth Factor beta2 / Q000235:genetics / Q000378:metabolism*; D055815:Young Adult
D018727:Muscarinic Antagonists; D012333:RNA, Messenger; D053781:Transforming Growth Factor beta2; D001285:Atropine; D002217:Carbachol