Study On The Effect And Mechanism Of Astragalus And Mulberry Bark On Myopic Scleral Remodeling Based On The Theory Of "White Eyes Belong To The Lung"

Objective To investigate the effects of Astragalus and Mulberry Bark on myopic scleral remodeling and the related mechanisms.Method1.Network pharmacology and molecular docking predict the role and mechanism of Astragalus and Mulberry Bark on myopic scleral remodeling.Through network pharmacology,the components,targets and signal pathways of Astragalus and Mulberry Bark on myopia sclera remodeling were analyzed,and the key components and targets were molecular docking.2.Experimental verification of the effect of Astragalus and Mulberry Bark on myopic scleral remodeling.The model of form deprivation myopia was constructed,and high,medium and low doses of Astragalus and Mulberry Bark were given respectively for intervention.The right eye diopter and axial length of guinea pigs were measured before and 2,4,6weeks after the model establishment;HE staining was used to observe the pathological changes of sclera,transmission electron microscopy was used to observe the ultrastructural changes of sclera,and Western Blot and RT-PCR were used to detect COL1 α 1 protein and m RNA expression.3.Experimental verification of the mechanism of Astragalus and Mulberry Bark in inhibiting myopic scleral remodeling.The model of form deprivation myopia was constructed,and high,medium and low doses of Astragalus membranaceus and Morus alba were given respectively for intervention.Six weeks later,the samples were taken and Western Blot,RT-PCR and immunofluorescence were used to detect the protein and m RNA expression of HIF-1α,α-SMA,and MMP-2.Result1.Network pharmacology and molecular docking predict the role and mechanism of Astragalus and Mulberry Bark on myopic scleral remodeling.(1)Network pharmacological analysis showed that quercetin,kaempferol,formononetin,3,9-di-O-methylnisolon,and 7-O-methylsomuconulatol were the key components of Astragalus in myopic scleral remodeling.IL-6,TNF,CASP3,MMP9,VEGFA,AKT1,EGFR,MMP2 and HIF1 A are the key targets of Astragalus for scleral remodeling in myopia.TNF signal pathway,HIF-1 signal pathway,MAPK signal pathway and PI3K-AKT signal pathway are the key signal pathways involved in the role of Astragalu in myopia sclera remodeling.Molecular docking showed that the binding energies of key components and targets involved in Astragalus acting on myopia sclera remodeling ranged from-5.2 to-8.6 kcal/mol,indicating that the affinity of active components and targets was good.(2)The network pharmacological analysis showed that quercetin,kaempferol,beta sitosterol,Iristectorigenin(9CI)and pelargonidin were the key compounds of Mulberry Bark on myopia sclera remodeling.The targets of IL6,TNF,AKT1,CASP3,MMP9,VEGFA,EGFR,MMP2 and HIF1 A are the key targets of Mulberry Bark on myopic scleral remodeling.TNF signal pathway,HIF-1α signal pathway and IL-17 signal pathway are the key signal pathways involved in Mulberry Bark’s effect on myopia sclera remodeling.The docking binding energies of the key active components and targets of Mulberry Bark on myopia scleral remodeling ranged from-6 to-9.9kcal/mol,indicating that the affinity of the key components and targets was good.2.Experimental verification of the effect of Astragalus and Mulberry Bark on myopic scleral remodeling.(1)The form deprivation myopia model was successfully constructed.Compared with FDM group,the myopic diopter and ocular axis of the right eye of the guinea pigs in the HQH and HQM groups increased slowly(P < 0.01);HE staining showed that compared with FDM group,the sclera of each dosage group of Astragalus was relatively thicker,the diameter of collagen fibers was relatively larger,the proportion of small diameter collagen fibers was lesser,and the gap of collagen fibers was smaller;The results of Western Blot and RT-PCR showed that compared with the FDM group,The expression of protein and m RNA of Co L1α1 in HQH group and HQM group was high(P<0.05).(2)The form deprivation myopia model was successfully constructed.Compared with the FDM group,the myopic diopter and ocular axis of the right eye of the guinea pigs in the SBPH group and the SBPM group increased slowly(P < 0.01);HE staining showed that compared with FDM group,the sclera of each dosage group of Mulberry Bark was relatively thicker,the diameter of collagen fibers was relatively larger,the proportion of small diameter collagen fibers was lesser,and the gap of collagen fibers was smaller;The results of Western Blot and RT-PCR showed that compared with FDM group,The expression of protein and m RNA of Co L1α1 in SBPH group and SBPM group was high(P<0.05).3.Experimental verification of the mechanism of Astragalus and Mulberry Bark in inhibiting myopic scleral remodeling.(1)In sclera tissue,Western Blot,RT-PCR and immunofluorescence results showed that compared with FDM group,the expression of protein and m RNA of HIF-1α,α-SMA and MMP-2 decreased in HQH group(P<0.05),the expression of protein and m RNA of MMP-2 decreased in HQM group(P<0.05).In choroidal tissue,Western Blot results showed that compared with FDM group,the protein expression of HIF-1αdecreased in HQH group and HQM(P<0.05).(2)Western Blot,RT-PCR and immunofluorescence results showed that compared with FDM group,the protein and m RNA expression of sclera HIF-1α and α-SMA did not change significantly in SBPH group,SBPM group and SBPL group(P>0.05),and compared with FDM group,the protein and m RNA expression of MMP-2decreased in SBPH group and SBPM group(P<0.05).Conclusions(1)Based on the theory that "white eyes belong to the lung",the occurrence and development of myopia are closely related to the lung.In the local syndrome differentiation of myopia,we can consider treating it from the lung.(2)Astragalus and Mulberry Bark can inhibit the scleral remodeling of myopia and delay the progress of form deprivation myopia.(3)Astragalus may play an inhibitory role in myopia sclera remodeling by promoting the synthesis of sclera collagen and inhibiting the degradation of sclera collagen;Mulberry Bark may inhibit the remodeling of myopia sclera by inhibiting the degradation of sclera collagen.Astragalus inhibits the transdifferentiation of scleral fibroblasts and promotes sclera collagen synthesis,which may be related to the improvement of scleral hypoxia;Astragalus and Mulberry Bark can down regulate matrix metalloproteinase-2 and reduce the degradation of sclera collagen,which may be related to the inhibition of inflammatory reaction.