[Pressure influence on mRNA and protein expression of inducible nitric oxide synthase in purified retinal ganglion cells of rats].
OBJECTIVE
To study the inducible nitric oxide synthase (iNOS) mRNA and protein expression in rat purified retinal ganglion cells (RGCs) that cultured under different pressures.


METHODS
(1) To culture RGCs that from Sprague Dawley (SD) neonatal rats (postnatal 1 - 5 days) on two planes in assimilative culture solution, RGCs were purified by Thy1.1 with sheep anti rat FITC monoclonal antibody. (2) RGCs were cultured under pressures of 0, 20, 40, 60 and 80 mm Hg, respectively. The changes of iNOS mRNA and protein in RGCs under different pressures were demonstrated qualitatively and quantitatively by in situ hybridization, RT-PCR and Western blot.


RESULTS
After cultured for 12 and 24 hours in vitro, the purification rate of RGCs in the experiment reached 98%. The expression of iNOS mRNA and protein in RGCs became higher and higher as the pressure was increased. There were no iNOS mRNA and protein expression in the control group (0 mm Hg) and weak expression in 20 mm Hg group (P < 0.05). 40, 60 and 80 mm Hg groups had a very significant difference from the control group, respectively (P < 0.01).


CONCLUSIONS
Pressure can evoke the expression of nitric oxide synthase mRNA and protein in purified retinal ganglion cells in vitro, thus the nitric oxide can be one of the causes of RGC damage, that may induce or aggravate glaucoma.
Hu
Zhulin
Z
Department of Ophthalmology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. hzl77@263.net
Du
Shuhua
S
D000399:Air Pressure; D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002478:Cells, Cultured; D015971:Gene Expression Regulation, Enzymologic; D019001:Nitric Oxide Synthase / Q000235:genetics* / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat