Effects Of Qingguangan Granules On The Expression Of Autophagy-related Proteins In The Retina And Optic Nerve Tissues Of DBA/2J Mic

Objective: To investigate the effects of Qingguang ’an granules on expressions of LC3,LAMP1,Caspase-3,Cyt C and Parkin in retina and optic nerve tissue of DBA/2J mice based on mitophagy.Methods: Twenty C57BL/6J mice were divided into optic nerve blank group and retinal blank group,with 10 mice in each group.Sixty DBA/2J mice were randomly divided into optic nerve model group,retinal model group,optic nerve low-dose group,retinal low-dose group,optic nerve high-dose group and retinal high-dose group,with 10 mice in each group.The mice were fed to 38 weeks of age,during which the intraocular pressure was observed,and the DBA/2J mice were given intragastric administration after automatic modeling.The model group was intragastric with normal saline every day.Low dose group and high dose group were given 20.75 and 103.75 g/kg of Qingguang,once a day,respectively.The mice in each group were fed with conventional diet.After 15 days of intragastric administration,the right optic nerve was taken from all optic nerve groups and the left retina was taken from all retinal groups.Western blot was used to detect the protein expressions of LC3,LAMP1,Caspase-3 and Cyt C,and RT-q PCR was used to detect the m RNA expressions of Parkin,LAMP1 and Caspase-3.Results: 1.Intraocular pressure: After feeding to 38 weeks of age,the intraocular pressure of DBA/2J mice increased,and the modeling was successful.2.The protein expressions of LC3,LAMP1,Caspase-3 and Cyt C in the optic nerve and retina of mice were detected by Western blot.In the optic nerve group,the protein expression levels of Caspase-3,Cyt C,LAMP1,LC3-Ⅱ/LC3-Ⅰ in the optic nerve model group were increased compared with the blank optic nerve group(P<0.05).Compared with optic nerve model group,protein expression levels of LAMP1,Cyt C,LC3-Ⅱ/LC3-Ⅰ in low-dose optic nerve group were decreased(P<0.05),and protein expression levels of LAMP1,Caspase-3,Cyt C,LC3-Ⅱ/LC3-Ⅰ in high-dose optic nerve group were decreased(P<0.05).In the retinal group,compared with the blank retinal group,the protein expression levels of Caspase-3,Cyt C,LC3-Ⅱ/LC3-Ⅰ in the retinal model group were increased(P<0.05).Compared with retinal model group,protein expression levels of Caspase-3,Cyt C,LC3-Ⅱ/LC3-Ⅰ in low-dose retinal group were decreased(P<0.05),and protein expression levels of Caspase-3,LC3-Ⅱ/LC3-Ⅰ in high-dose retinal group were decreased(P<0.05).3.The m RNA expressions of Parkin,Caspase-3 and LAMP1 in the optic nerve and retina of mice were detected by RT-q PCR.In the optic nerve group,compared with the blank optic nerve group,m RNA expression levels of Parkin,Caspase-3 and LAMP1 in the optic nerve model group were increased(P<0.05).Compared with the optic nerve model group,the m RNA expression levels of LAMP1 and Caspase-3 in the low-dose optic nerve group were decreased(P<0.05),and the m RNA expression levels of Parkin,Caspase-3 and LAMP1 in the high-dose optic nerve group were decreased(P<0.05).Compared with low-dose group,m RNA expression levels of Parkin,Caspase-3 and LAMP1 in high-dose group were decreased(P<0.05).In the retina group,compared with the blank retina group,the expression level of Parkin m RNA in the retinal model group was decreased(P<0.05).Compared with retinal model group,Parkin m RNA expression level was increased in retinal high-dose group(P<0.05).Compared with low-dose retinal group,LAMP1 m RNA expression level in high-dose retinal group was decreased(P<0.05),and Parkin m RNA expression level was increased(P<0.05).Couclusion: 1.By regulating the expressions of Parkin,LAMP1,LC3,Caspase-3 and Cyt C in the optic nerve of glaucoma mice,Qingguang ’an Granules can reduce the accumulation of autophagy,make the autophagy process of mitochondria complete,and have a protective effect on the optic nerve.2.Qingguang ’an Granules can reduce the protein expressions of LC3-II/LC3-I and Caspase-3 in the retina of glaucoma mice to a certain extent,increase the expression of Parkin m RNA,strengthen the signal of mitochondrial autophagy,reduce the amount of excess mitochondrial autophagy in the retina of glaucoma mice,and help to reduce the accumulation of autophagosomes.Achieve the purpose of protecting the retina.