Effects Of Liver And Kidney Tonifying Therapy On The Expression Of HA,CD44 And C-jun In Scleral Fibroblasts Of Mouse Form Deprivation

Objective: To investigate the effects of liver and kidney tonifying therapy on the expression of hyaluronic acid(HA),CD44 and C-jun in scleral fibroblasts of C57BL/6J mice with form deprivation myopia and the molecular mechanism of fibroblast apoptosis.To provide experimental data for the application of liver and kidney tonifying method in delaying the development of myopia,and provide more theoretical basis for the treatment of myopia in traditional Chinese medicine.Methods:Normal healthy mice were randomly assigned to control group 1,model 1group,Chinese medicine intervention group,control group 2,model 2 group,Chinese medicine high,medium and low dose groups and positive drug control group.The model group and the drug-administered group(including the Chinese medicine intervention group,the Chinese medicine high,medium and low dose groups and the positive drug control group)were subjected to form deprivation with a translucent goggle in the right eye.The expressions of HA,CD44 and c-Jun in sclera of mice in each group were observed by immunohistochemical method.The contents of HA,CD44 and c-Jun in sclera of mice in each group were evaluated by Western blot.QRT-The expression of Has2 mRNA,CD44 mRNA and Jun mRNA in the sclera of each group was detected by PCR.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.Immunohistochemical results: In the first and second phases of experiment,the average optical density of HA protein in model 2 group was significantly lower than that in control group 2(P<0.01).The drug-administered group(Chinese medicine intervention group,Chinese medicine high,medium and low dose groups,positive drug control group)The average optical density of HA protein increased compared with the model group(P<0.05);The results of CD44 immunohistochemistry were not statistically significant(P>0.05),but the average optical density of CD44 proteindecreased in the model group.The drug-administered group(Chinese medicine intervention group,Chinese medicine high,medium and low dose groups,positive drug control group)Group)The average optical density value of CD44 protein has a decreasing trend compared with the model group;The mean optical density of C-jun in the model group 1 was not significantly different from that in the control group(P>0.05),but the average optical density of the C-jun in the sclera of the model group1 was higher than that in the control group 1.Model 2 The average optical density of C-jun protein in group was significantly higher than that in control group 2(P<0.01).The average optical density of C-jun protein in the drug-administered group(Chinese medicine intervention group,high-and low-dose Chinese medicine group)decreased compared with the model group.P<0.05).3.Western blot:In the first and second phases of the experiment,the results of HA,CD44 and C-jun Western blot were not statistically significant(P>0.05),but the protein content of HA in model 2 group was lower than that in control group 2,high and medium doses of traditional Chinese medicine.Compared with the model 2 group,the target protein content of HA increased;Compared with the control group,the content of the target protein in the model group decreased.The high-and medium-dose groups of the Chinese medicine group had an increasing trend compared with the model group 2;the protein content of the C-jun in the sclera of the model group 1 increased compared with the control group 1 and the Chinese medicine intervention group C The protein content of-jun had a decreasing trend compared with the model group 1.The protein content of C-jun in the high-and medium-dose groups of the Chinese medicine group and the positive drug control group increased compared with the model group 2.4.QRT-PCR:Experiment phase 1 and experiment phase 2,there was no significant difference in the results of QRT-PCR between the control group 1 and the model group 1 and the Chinese medicine intervention group(P>0.05),but the Has2 mRNA gene expression was down-regulated in the model 1 group.The expression of Has2 mRNA in model 2 group was significantly lower than that in control group 2(P<0.01);There was no significant difference in the expression of HA target geneHas2 mRNA between the high-,medium-,and low-dose groups of Chinese medicine and the control group(P>0.05),but there was an up-regulation trend.The expression of CD44 mRNA in the low-dose group of Chinese medicine was higher than that in the model 2 group.(P<0.05);The results of QRT-PCR of JUN mRNA gene were not statistically significant(P>0.05),but the expression of JUNmRNA in model 2 group was up-regulated compared with control group 2,the drug-administered group(high-,medium-,and low-dose groups of Chinese medicine and positive drug control group).There is a downward trend.Conclusion:1.Formal deprivation can induce relative myopia in C57BL/6J mice,and the induced myopic diopter and axial length are proportional to the length of the form deprivation.2.During the development of myopia,HA and CD44 were down-regulated in scleral tissue and c-jun expression was elevated.During myopia intervention,HA expression was increased in sclera and c-jun was down-regulated.3.The liver and kidney supplement Chinese medicine Bujingyishi tablets may achieve the purpose of intervention for myopia by up-regulating the expression of HA in myopia and down-regulating the expression of c-jun.