Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-asso...Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.展开更多
AIM To discuss the expression of endotheline and alpha smooth muscle actin in periretinal membrane of proliferative vitreoretinopathy (PVR). METHODS 14 periretinal membranes (PCM) obtained from 14 cases of PVR withret...AIM To discuss the expression of endotheline and alpha smooth muscle actin in periretinal membrane of proliferative vitreoretinopathy (PVR). METHODS 14 periretinal membranes (PCM) obtained from 14 cases of PVR withretinal detachment (RD) undergoing vitretomy were used to examine the expression of endotheline 1 (ET 1) and α SMA by immunohistochemical detection. RESULTS The reactions of ET 1 were observed in the cells and extracellular matrix (ECM) in the various grades of the PRMs and darken at the rim of the PCM, in which some of cytonucleus became the shape of fusiform and/or slimer and longer. While α SMA was expressed in the centre and verge of various grades the PCM, and the positive cells were parallel to the contractive orientation of the PCM. CONCLUSION Both ET 1 and α SMA could be expressed in the PCM, the cells in the PRMs may express ET and secret it to ECM and deliver effects on the contractive cells that express α SMA.展开更多
基金supported by the National Natural Science Foundation of China,Nos.82071008(to BL)and 82004001(to XJ)Medical Science and Technology Program of Health Commission of Henan Province,No.LHGJ20210072(to RQ)Science and Technology Department of Henan Province,No.212102310307(to XJ)。
文摘Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.
文摘AIM To discuss the expression of endotheline and alpha smooth muscle actin in periretinal membrane of proliferative vitreoretinopathy (PVR). METHODS 14 periretinal membranes (PCM) obtained from 14 cases of PVR withretinal detachment (RD) undergoing vitretomy were used to examine the expression of endotheline 1 (ET 1) and α SMA by immunohistochemical detection. RESULTS The reactions of ET 1 were observed in the cells and extracellular matrix (ECM) in the various grades of the PRMs and darken at the rim of the PCM, in which some of cytonucleus became the shape of fusiform and/or slimer and longer. While α SMA was expressed in the centre and verge of various grades the PCM, and the positive cells were parallel to the contractive orientation of the PCM. CONCLUSION Both ET 1 and α SMA could be expressed in the PCM, the cells in the PRMs may express ET and secret it to ECM and deliver effects on the contractive cells that express α SMA.
