| Objective:Hyaluronic acid(HA)plays an important role in the development of the retina.CD44 plays an important role in regulating the adhesion of matrix components between the neural retina and photoreceptors.We aimed to investigate whether HA/CD44 has an effect on the retina of experimental myopia mice and whether it has a regulatory effect on liver and kidney supplementation,in order to guide early clinical intervention and prevent or delay the occurrence of high myopia retinal atrophy.Methods:In this experiment,We chose a total of 180 healthy C57BL/6J mice of 3 weeks old.The model was used to cover the right eye of C57BL/6J mice with a translucent eye mask to prepare the form deprivation myopia model(except the control group).For the experimental eye comparison,the experiment was carried out in two stages: the mice were examined by the retinoscopy optometry after the optometry.Phase I of the experiment: According to the random number table method,180 mice were randomly divided into 3 groups: 40 in the control group(no treatment in both eyes)and 120 in the model group(the 22 nd day-old right eye began to form deprivation)20 Chinese medicine intervention group(22nd day old right eye shape deprivation and Chinese medicine gavage);Chinese medicine intervention group was given a supplement of Bujing Yishi Tablet with liver and kidney supplement effect,the dosage was 0.50 g /kg/day,the control group and the model group were all intragastrically administrated with the same amount of distilled water,once a day.The mice were re-examined and optometry after the mice were raised to 43 days of age,and the rats in the intervention group were randomly sacrificed.Twelve mice in the control group and the control group were labeled as control group 1 and model 1 group.All right mice were removed and the eye axis was measured with a digital micrometer.The second phase of the experiment: the remaining control mice were labeled as control 2 groups(both eyes did not do any treatment),the model group mice continued to form deprivation in the right eye,and were randomly divided into 5 groups: model 2 group,low Chinese medicine,In the middle dose groups and high dose groups,the positive control group;the low,medium and high dose groups of traditional Chinese medicine were administered with the supplemental solution of Bujing Yishi Tablet with Liver and kidney with filling,and the dosage was 0.25g/kg/day,0.50g/kg/day,1.00g/kg/day,the positive drug control group was given a suspension of mecobalamin tablets,the dose was 0.20mg/kg/day,and the control group 2 and the model 2 group were used.The distilled water was intragastrically administered once a day,and all the mice in this part were sacrificed to death at 64 days of age.The diopter of each group was measured by band photodetector before and after the experiment,.The mice after sacrifice were measured by the digital micrometer to measure the axial length of each group.The immunohistochemical method was used to observe the retina of each group after the experiment.HA,CD44,IAP,c-Jun protein expression in tissues;Western blot analysis of HA,CD44,c-Jun,IAP(Immunosuppressive Acidic Potein,IAP)protein content in retinal tissue of each group;QRT-PCR The expression of Has2 m RNA,CD44 m RNA,Junm RNA and Birc3 m RNA in retinal tissue of mice in each group after the test.Results:1.Refraction and Axis Length: Form deprivation can reduce the diopter of mice and increase the length of the axial length.The liver and kidney supplement Chinese medicine Bujing Yishi Tablet can control the drift of mouse diopter to myopia and slow down the length of the axial length.2.Immunohistoche mical results: In the first phase of experiment and the second phase of experiment,the average optical density of HA and IAP protein in the model group decreased compared with the control group.The drug-administered group(Chinese medicine intervention group,Chinese medicine high,medium and low dose groups and The positive optical density values of HA and IAP proteins in the positive drug control group were higher than those in the model group;the average optical density values of c-Jun protein in the model group were higher than those in the control group,and the drug-administered group(Chinese medicine intervention group,Chinese medicine high,medium and low dose groups and The positive optical density values of HA and IAP proteins in the positive drug control group were lower than those in the model group;the CD44 immunohistochemical test results were not statistically significant(P>0.05),and the CD44 was in the model group and the drug-administered group(Chinese medicine intervention group,Chinese medicine was high,The middle and low dose groups and the positive drug control group all had a downward trend.3.Western blot:At the first stage of experiment,the contents of HA,CD44 and IAP in model group 1 were lower than those in control group.The contents of HA,CD44 and IAP in Chinese medicine intervention group were higher than those in model group 1.The content of c-Jun protein in model group 1 was higher than that in control group 1.There was a downward trend,and the c-Jun protein content model of the retinal tissue of the Chinese medicine intervention group was elevated.In the second phase of experiment,the protein content of HA,CD44 and IAP in model 2 group decreased compared with the control group 2,and the protein content of HA,CD44 and IAP in the drug-administered group increased compared with the model 2 group;the model 2 group had the c-Jun protein content in the control group 2 The upward trend,the content of c-Jun protein in the retinal tissue of the high,middle and low dose groups of Chinese medicine and the positive drug control group increased compared with the model 2 group.4.Quantitative RT-PCR:At the first stage of experiment,the expressions of Has2 m RNA,CD44 m RNA and Birc3 m RNA in model group 1 were down-regulated compared with control group 1.The expressions of Has2 m RNA,CD44 m RNA and Birc3 m RNA in Chinese medicine intervention group were significantly up-regulated compared with model group 1;the expression of Junm RNA in model group 1 was higher than that in control group 1 The group showed a significant upward trend.The expression of Junm RNA in the Chinese medicine intervention group was significantly lower than that in the control group.In the second phase of experiment,the expressions of Has2 m RNA,CD44 m RNA and Birc3 m RNA in model 2 group were down-regulated compared with the control group.The expressions of Has2 m RNA and CD44m RNA in the middle dose group and the low dose group were up-regulated compared with the model group 2.The high-dose group of traditional Chinese medicine The expression of Has2 m RNA in the control group was up-regulated compared with the model 2 group.The expression of CD44 m RNA in the high-dose group and the positive control group was down-regulated compared with the model 2 group.The expression of Birc3 m RNA in the drug-administered group was up-regulated compared with the model 2 group.The expression of Junm RNA in model 2 group was down-regulated compared with control group 2,and the expression of Junm RNA in retinal tissue of high-,medium-,and low-dose groups of Chinese medicine was higher than that in model 2 group.Conclusion:1.C57BL/6J mice can induce relative myopia by form deprivation,and the diopter and axial length of myopia increase with the prolongation of form deprivation time.2.In the process of myopia formation and myopia formation,HA,CD44 and IAP were down-regulated in the retinal tissue;HA,CD44 and IAP were up-regulated in the retinal tissue during the intervention of the liver and kidney supplement Chinese medicine Bujingyishi tablets;The expression of HA,CD44 and IAP in the middle dose group was significantly higher than that in the other dose groups.3.In the process of myopia formation and after the formation of myopia,the liver and kidney supplement Chinese medicine Bujing Yishi Tablet has a two-way regulation effect on c-Jun.4.liver and kidney with Chinese medicine Fujing Yishi tablets may up-regulate the expression of myopia retinal HA / CD44 to achieve the purpose of intervention for myopia. |
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