| ObjectiveTo investigate the effects of QimingWan on guinea pigs with form-deprivation myopia.The changes of myopic diopter and axial length of guinea pigs were measured,in order to demonstrate the effect of QimingWan on myopia progress.Meanwhile,the effects of QimingWan on the scleral tissue of myopia progress were preliminary studied by observing the scleral structures with HE,and measuring the relative molecular expression levels of MMP-2,TIMP-2 and TGF-?1 in scleral tissue.Method1.Establishment of FDM model:Forty 3-week-old guinea pigs were randomly divided into four groups:the high-dose Chinese herbal medicine group,low-dose Chinese herbal medicine group,model control group and blank control group,with 10 in each group.FDM model was established on the right eye of guinea pigs in the first three groups for four weeks,and served the left eye as control.Then all the guinea pigs were underwent to retinoscopy in the first three groups to confirm that myopia were induced.2.Drug intervention:The FDM model was kept on inducing for four weeks on the high-dose Chinese herbal medicine group,the low-dose Chinese herbal medicine group and the model control group after the myopia was induced,during the same time,all guinea pigs were intragastrically administered once a day for four weeks.The high-dose Chinese herbal medicine group and the low-dose Chinese herbal medicine group were administered with different concentrations of QimingWan;the model control group with an equal dose of saline;the blank control group with no treatment.3.Observation index:(1)Biological detection:All guinea pigs were underwent measurements(diopter,axial length)at the beginning of the experiment,at the end of the 4th week and at the end of 8th week;(2)Histopathology and molecular biology detection:All the guinea pigs were sacrificed and the eyeballs were taken for experimental operation at the end of 8th week.Some paraffih sections of the eyeballs were made and stained with eosin-hematoxylin.Then the differences of scleral collagen fibers were observed between different groups.The relative molecular expression of MMP-2,TIMP-2 and TGF-?1 in sclera was detected by WB.Meanwhile the ratio of MMP-2/TIMP-2 relative molecular expression was calculated.Result1.FDM model:(1)Evaluation of guinea pigs:There was no difference between the high-dose Chinese herbal medicine group,the low-dose Chinese herbal medicine group,the model blank group,and the blank control group in the diopter and axial length at the begin of the experiment(P>0.05).(2)Establishment of myopia model:The right eyes of the high-dose group,the low-dose group,and the model blank group were blocked with latex balloon masks for four weeks,then the diopter of all the guinea pigs were detected.The diopter of each model group showed a negative growth,and the diopter change averaged-3.82D.The changes of diopter before and after modeling in each model group and blank control group were compared,and the difference was highly statistically significant(P<0.05).It suggested that the FDM model in the guinea pigs were successfully established.2.Intervention of QimingWan on the diopter and axial length:(1)Changes of diopter:The guinea pigs were under the continuous modeling of each group.During the same time,the high-dose Chinese herbal medicine group and the low-dose Chinese herbal medicine group were intragastrically administered with different concentrations of QimingWan,and detected the diopter at the end of the 8th week.The diopter progression of the high-dose Chinese herbal medicine group together with the low-dose Chinese herbal medicine group was lower than that of the model control group.There was no significant difference between the high-and low-dose Chinese herbal medicine groups(P>0.05).The difference of diopter between left and right eyes in the model control group was higher than that in the high-and low-dose Chinese herbal medicine group.The differences between the three groups were statistically significant(P<0.05).There was no significant difference between the high-and low-dose Chinese herbal medicine group(P>0.05).(2)Changes of axial length:The changes of axial length growth of the high-dose Chinese herbal medicine group and the low-dose Chinese herbal medicine group were lower than that of the model control group.The differences were statistically significant(P<0.05).There was no statistical difference of the axial length between the high-and low-dose Chinese herbal medicine group(P>0.05).The difference of the axial length between the the left and right eyes of each group,the model control group was higher than that of the high-and low-dose Chinese herbal medicine groups.The difference between the three groups was statistically significant(P<0.05).There was no statistical difference between the high-and low-dose Chinese herbal medicine group(P>0.05).3.Histopathological manifestations of guinea pigs sclera:Paraffin sections of sclera were prepared after eight weeks of the experiment,and the scleral tissues of guinea pigs were observed under a light microscope.(1)Blank control group:The collagen fibers were closely arranged,and the bonding between the fibers showed a coexistence of lamellar and interlaced states.The arrangement of collagen fibers in the layers of the sclera was very neat.(2)Model control group:The structure of the sclera was loose,the gap between the collagen fibers was increased,the arrangement was sparse,and the phenomenon of fracture and separation occured,and the thickness of the sclera was thin,compared with the blank control control group.(3)High-and low-dose Chinese herbal medicine group:The interlaminar structure of the collagen fibers was clear,and the connection between the collagen fibers was mainly in the lamellar state,and less interlaced compared with the model control group.The thickness of the sclera was roughly equal to that of the blank control group,which was thicker than the model control group.4.Effect of the QimingWan on the expression of scleral remodeling related factor protein:Extraction of scleral tissue proteins after eight weeks of the experiment:(1)MMP-2,TIMP-2:The relative molecular expression of MMP-2 was the largest in the model control group than the others,and the difference of the relative molecular expression of MMP-2 was statistically significant(P<0.05).The relative molecular expression of MMP-2 in the high-and low-dose Chinese herbal medicine groups was lower than that in the model control group,and the difference was statistically significant(P<0.05).The ratio of MMP-2/TIMP-2 relative molecular expression in the model control group was significantly higher than that in the other three groups,and the difference was statistically significant(P<0.05).(2)TGF-?1:The relative molecular expression of TGF-?1 in the model control group and the high-and low-dose Chinese herbal medicine groups was higher than that in the blank control group,and the difference was statistically significant(P<0.05).There was no significant difference of the relative molecular expression of TGF-?1 between the high-and low-dose Chinese herbal medicine groups and the model control group(P>0.05).Conclusion1.The results suggest that QimingWan can reduce the progression of myopic diopter,delay the growth of the axial length,and affect the progression of FDM in guinea pigs.2.QimingWan balance the expression of MMP-2 and TIMP-2 by regulating the expression of MMP-2,inhibit the pathological degradation of scleral extracellular matrix in FDM guinea pigs.The guinea pig scleral remodeling is affected may be a possible mechanism for QimingWan to affect the progression of myopia.But the specific pathways need to be further researched. |
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