Influence Of Pressure On Retinal Ganglion Cells In Vitro And Protective Effects Of EBHM On Injury Of Retinal Ganglion Cells By Pressure

Research background:Glaucoma is a neuro degenerative disease characterized by progressive loss of Retinal Ganglion Cells and optic nerve axons,and accompanied by loss of visual field sensitivity.lt is the global second blinding eye disease, and is only second to cataract. Many factors besides intraocular pressure associated with optic nerve injury. Erigeron Breviscapus (Vant.) Hand Mazz is a kind of Chinese herbal medicine,It can improve microcirculation,and protect of glaucoma optic nerve.Numerous studies have shown that,it is ability to protect optic nerve on rats with experimental glaucoma. But the specific mechanism is still unclear.This experiment through the establishment of Retinal Ganglion cells high-pressure culture model in vitro,and use Erigeron Breviscapus (Vant.) Hand Mazz for Protective treatment,Discuss the possible mechanism of protective action.Purpose:1 To establish high pressure develop model of Retinal Ganglion Cells in vitro,and to observe the influence of pressure on Retinal Ganglion Cells;2 To observe the effect of Erigeron Breviscapus (Vant.) Hand Mazz on apoptosis of rat Retinal Ganglions Cells cultured in vitro,and To discuss the possible mechanism of its neuroprotective effectAnd to provide more theoretical basis on cell and molecular level for the neuroprotective effect of Erigeron Breviscapus (Vant.) Hand Mazz in ophthalmology.Methods:1 establish high pressure culture model of Retinal Ganglion Cells in vitro.2 After 24 hours,in mixed culture of nerve cells put the lamps of different concentrations of Erigeron Breviscapus (Vant.) Hand Mazz、Nerve growth factor(NGF)、culture, then test the OD of living eells by MTT after anhter 24 hours.3 After 24 hours, in mixed culture of Retinal Ganglion Cells put the lamps of different concentrations of Erigeron Breviscapus (Vant.) Hand Mazz、Nerve growth factor(NGF)、culture, and according to the experimental design, using a special device for experimental group and model group sealed pressure to 80 mmHg,4hours,continue to develop for 24 hours, then measure the glutamic acid and γ-aminobutyric acid content in the supernatant with Elisa method.Results:1 RGCs mixed culture can survive longest 2-3 weeks.2 Pressure can promote apoptosis of RGCs.3 In the training of the RGCs, the OD value of 1 mg/ml,0.1 mg/ml concentration of Erigeron Breviscapus (Vant.) Hand Mazz and Nerve growth factor(NGF)is higher than the model group (P<0.05).4 Model group RGCs under the condition of 80 mmHg pressure train 4 hours,its glutamic acid content in the supernatant fluid is higher than blank control group,and γ-aminobutyric acid content is also higher than blank control group (P<0.05).Glutamic acid content in the supernatant fluid of Erigeron Breviscapus (Vant.) Hand Mazz group is lower than the model group (P<0.05), γ-aminobutyric acid content in the supernatant fluid of Erigeron Breviscapus (Vant.) Hand Mazz group is higher than the model group (P<0.05)Conclusions:1 Pressure can promote RGCs cultured in vitro apoptosis.2 Erigeron Breviscapus (Vant.) Hand Mazz has protective effect on RGCs cultured in vitro and can against apoptosis of RGCs by pressure,The mechanism may be by reducing the cells release of excitatory toxin-glutamate,Increasing the cells release of γ-aminobutyric acid to against the excitatory toxic effect of glutamate.