| Objective:Preliminary discussion the effects of liver and kidney supplementation on refractive error,axial length,retinal tissue structure and retinal cell apoptosis in form deprivation myopia(FDM)C57BL/6J mice.Provide experimental basis and new research ideas of early visual function protection in high myopia.Methods:The experiment was divided into two parts.63 3 weeks old healthy C57BL/6J mice were selected to enter the two parts of the experiment.The two part of the experiment did not share the mice.The experimental model of form deprivation myopia(except the normal control group)was made by using the translucent eye mask to cover the right eye of the right eye(except the normal control group).All the groups were compared with the right eye as the experimental eye.The following is as follows:Part I:21 three-week-old healthy C57BL/6J mice were selected for experiment.The experimental period was 3 weeks.They were rand omly divided into3 groups: 3-week normal control group(uncovered of both eyes)and 3 weeks model group(right eye deprivated 3 weeks),traditional chinese medicine intervention group(right eye form deprivation at the same time intragastric administration the TCM 3weeks),6 mice in each group,The intervention group of TCM was given intragastrically with Bujin yishi Tablets suspension with the same effect of liver and kidneys,intragastric volume 0.1ml/10 g,and concentration 54.6mg/ml;the normal control group and 3-week model group were fed with equal amount of distilled water.The mice were sacrificed at 43 days of age.Part II:42 three-week-old healthy C57BL/6J mice were selected for experiment.The experimental period was 6 weeks.They were randomly divided into 6 groups: 6-week normal control group(without any treatment in both eyes)and 6 weeks model group(Right eye deprivated 6 weeks),low,medium and high dose group in TCM and positive drug control group(4 groups were formally deprived for 6 weeks,After 3 weeks of deprivation,mice were administrated for 3 weeks),6 in each group,The low,medium,and high doses of TCM were intragastrically administered with Bujin yishi Tablets with the same effect of liver and kidneys.The volume of intragastric administration was 0.1 ml/10 g,andthe concentrations were: 27.3 mg/ml,54.6 mg/ml,and 109.2 mg/ml respectively..The positive drug control group was given a mecobalamin tablet suspension gavage with a volume of 0.01 mg/ml and a concentration of 0.1 ml/10 g.The 6-week normal control group and 6-week model group were fed with equal amount of distilled water.All administration groups were administered at a same daily time.The mice were sacrificed at 64 days of age.Before and after the two parts of the experiment,the refraction of each group was measured by a same optometrist with band-shaped light retinoscope.After the experiment,all mice’s eyeball enucleated and their axial length measured with a digital micrometer;Retinal HE staining was used in histopathology of mice in each group under light microscope.The TUNEL method was used to observe the apoptosis of retinal cells.The difference between the experimental eyes and the normal control eyes was compared.Results:1.Refraction and axial length:There was no significant difference in refraction before experiment between experimental group 1 and experimental group 2(P>0.05).(1)Experiment I: The refractive power and axial length of the experimental group in the 3-week model group were significantly different from those in the 3-week normal control group and the TCM intervention group(P<0.05).(2)Experiment 2: The refractive power and axial length of experimental eyes in the 6-week model group were significantly different from those in the 6-week normal control group,Chinese medicine,low-dose,high-dose group,and positive drug control group(P<0.05).The refraction and axial length of the high,low,medium dose,and positive drug control groups were lower than those in the 6-week model group(P < 0.05);there was no significant difference between the Chinese medicine low-dose group and the positive drug control group(P> 0.05).2.HE staining:The retina structure was clear and the level was clear in the 3-weeks normal control group and the 6-week normal control group.There was no significant difference in the thickness of retinal pigment epithelium and photoreceptor layer between experiment 1 and experiment 2(P>0.05).(1)Experiment I: The retinal thickness of the experimental group in the 3-week model group was thinner than thatin the 3-weeks normal control group and the Chinese medicine intervention group(P<0.05),mainly in the inner and outer nuclear layers and nerve fiber layers.The number of cells in the inner and outer nuclear layers decreased,the arrangement was sparse,and the outer segments of the visual cell layer were disorderly arranged.The thickness of inner nuclear layer,outer nuclear layer,thickness of nerve fiber layer,number of inner and outer nuclear layer cells(mean blackness average)and number of ganglion cells in the model group were significantly different from those in the three-week normal control group and the traditional Chinese medicine intervention group(P<0.05).).(2)Experiment 2: The retinal thickness became thinner in the6-week model group than in the high,middle,and low dose groups of the Chinese medicine,the positive drug control group,and the 6-week normal control group(P<0.05),mainly in the inner and outer nuclear layers and nerve fiber layers..The number of cells in the inner and outer nuclear layers decreased,the arrangement was sparse,and the outer segments of the visual cell layer were disorderly arranged.The thickness of inner and outer nuclear layer,nerve fiber layer thickness,number of inner and outer nuclear layer cells(average mean blackness),and number of ganglion cells in the high-dose group were significantly different from those in the middle and low dose group and the positive drug control group(P<0.05).<0.05),but there was no significant difference between the Chinese medicine low dose group and the positive drug control group(P> 0.05)3.TUNEL:No apoptotic cells were detected in the retina of the normal control group at 3 and 6 weeks.In experiment 1 and experiment 2,except for the normal control group,cells stained with positive TUNEL reagents were seen in the retinas of the other groups.The nucleus was brownish yellow.Apoptotic cells were mainly distributed in the inner nuclear layer,outer nuclear layer,and ganglion cell layer.The largest number of apoptotic cells was found in layers and ganglion cell layers.Compared with the 3 week model group,the number of retinal cell apoptosis in Chinese medicine intervention group was significantly different(P<0.05).There was significant difference between the 6 weeks model group and the other 6 weeks(P<0.05).There was significant difference in the retinal cell apoptosis between thehigh dose group and the low dose group and the positive drug control group(P<0.05).There was no significant difference between the Chinese medicine and the low dose group and the positive drug control group(P >0.05).Conclusion:1.Form deprivation can induce C57BL/6J mice to induce myopia and ocular axis elongation to FDM.With the prolongation of form deprivation time,the diopter and axial length of myopia induced by mice also increase gradually.2.The formation of myopia has a certain effect on the structure of the retina.The liver kidney and kidney supplementation have some intervention on the formation and development of experimental myopia and the morphology of retina.3.Apoptosis of retina cells exists in the development of myopia.Hepatorenal supplementation has a certain intervention effect on the apoptosis of myopic retinal cells in C57BL/6J mice. |
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