The Effects Of Chinese Herbal Recipe Huoxue Huayu Decoction On Retinal Glutamate Transporter GLAST In Acute Ocular Hypertension Of Rats

ObjectiveTo explore the protection effects of huoxue huayu decoction(HXHYD) on retinal ganglion cells(RGCs) in acute ocular hypertension model of Wistar ratsMethodsForty-five healthy Wistar rats were divided into three groups,group A,B and C randomly.Group A was normal group which included 5 rats.Group B and C,each group included 20 rats,either eye of those rats was operated acute hypertension model surgery. Group C received HXHYD(4ml,oral,one time per day) after surgery,5 rats were executed randomly at 1,3,7and 14 days after the surgery.Group B did the same measure as Group C.Expression of retinal glutamate transporter GLAST of acute ocular hypertension and normal rat retina were detected through immunofluorohistochemistry and real-time quantitative polymerase chain reaction.Those rats' retinas were observed by light microscopy and electron microscopy.Results1,Light microscope:After 3 days of acute intraocular hypertension,the retina became thin,RGCs numbers became decrease,after 7 days of acute intraocular hypertension, the retina became thinner,particularly in the inner plexiform layer,RGCs numbers reduce distinctly;after 14 days of acute intraocular hypertension,the above changes became more significant,the inner and outer nuclear layers were sparseness and cells numbers were decreased.The damage of treated group was slighter than pressurizing group.2,Transmission electron microscopy:After 1 days of acute intraocular hypertension,the retinal ganglion cell chromatin was gathered to the surrounding,the organelle were degradation,mitochondria was degeneration,endoplasmic reticulum and lysosome were swelling;after 14 days of acute intraocular hypertension,the retinal ganglion cells were degeneration seriously,the cell nuclear were dissolved,mitochondria and endoplasmic reticulum were swelling significantly.The trauma of treated group was slighter than pressurizing group.3,Fluorescence immunohistochemistry:(1) pressurizing group:GLAST was increased rapidly compared with normal group after 1 day(P＜0.01) of acute intraocular hypertension,descended at 3 days (P＜0.05),reverted to the normal group level at 7 days(P＞0.05).(2) Treated group:In comparison with normal group,GLAST was increased rapidly after 1 day(P＜0.01) of acute intraocular hypertension,descended at 3 days(P＜0.01).In comparison with pressurizing group,GLAST was increased after 1 day(P＜0.05) of acute intraocular hypertension,descended at 3 days (P＜0.05).After 7 days of acute intraocular hypertension,there was no significant change compared with normal and pressurizing group(P＞0.05).4,real-time quantitative polymerase chain reaction(qRT-PCR):(1) pressurizing group:The expression of GLASTmRNA was increased rapidly compared with normal group after 1 day(P＜0.05) of acute intraocular hypertension,descended at 3 days(P＜0.01),reverted to the normal group level at 7 days(P＞0.05).(2) Treated group:In comparison with normal group,the expression of GLASTmRNA was increased rapidly after 1 day(P＜0.01) of acute intraocular hypertension,there was no significant change at 3 days(P＞0.05).In comparison with pressurizing group,the expression of GLASTmRNA was increased after 1 day(P＜0.05) of acute intraocular hypertension,the expression of GLASTmRNA was higher than pressurizing group at 3 days(P＜0.05).After 7 days of acute intraocular hypertension,there was no significant change compared with normal and pressurizing group(P＞0.05).Conclusion1,After acute intraocular hypertension,the retina became thin,especially the inner layers,and RGCs numbers were decreased,parts of cells were apoptosis.2,GLAST was significantly increased in both pressurizing and treated group after acute intraocular hypertension,and then decrease,reverted to the normal group level eventually.Compared with pressurizing group,the increase degree of treated group was higher,and the decrease degree was lower,the time that recovered to the normal level was shorter.3,HXHYD's protective effect on retinal ganglion cells in rats acute intraocular hypertension model which is probably because HXHYD can elevate the expression of Glutamate transporters in Muller cells,especial GLAST.