[Inhibition of IL-6 antisense oligonucleotide on IL-6 expression by human retinal pigment epithelium in vitro].
OBJECTIVE
To investigate the inhibition of IL-6 antisense oligonucleotide (ASON) on IL-6 expression by retinal pigment epithelium (RPE) cells on the basis of previous study.


METHODS
Cultured human RPE cells was treated with IL-1 beta and IL-6 ASON. IL-6 mRNA and protein expression were detected by enzyme linked immunosorbent assay (ELISA), immunohistochemistry and in situ hybridization histochemistry (ISH).


RESULTS
It was demonstrated that the IL-6 expression by RPE cells was dose and time dependent after the stimulation of IL-1 beta. IL-6 in the conditioned media or in the control group was 2.00 x 10(-6) g/L cells after the exposed to IL-1 beta for 8 hours. IL-6 ASON (2.00 x 10(-5) mol/L) obviously inhibited IL-6 (5.50 x 10(-7) g/L cells, t = 4.518, P < 0.01) production. Immunohistochemistry study showed dark blue staining in RPE cytoplasm after stimulation with IL-1 beta, while the cells treated with IL-6 ASON showed light staining in RPE cytoplasma. ISH displayed that there was a marked reduction in mRNA expression in IL-6 ASON treated group compared with that in the control group (t = 3.746, P < 0.01).


CONCLUSIONS
Cultured RPE cells express IL-6 protein and mRNA in dose and time dependent manners when RPE cells are stimulated with IL-1 beta. The expressions of IL-6 protein and mRNA can be significantly inhibited by IL-6 ASON in cultured human RPE cells.
Song
Zongming
Z
Department of Ophthalmology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. songzongming@263.net
Hui
Yannian
Y
Qu
Jia
J
Wang
Yusheng
Y
Ma
Jixian
J
Si
Yanfang
Y
Kang
Jun
J
D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D013997:Time Factors; D000093242:Interleukin-6 Inhibitors
D007375:Interleukin-1; D015850:Interleukin-6; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D000093242:Interleukin-6 Inhibitors