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Effect of Sonic hedgehog gene-modified bone marrow mesenchymal stem cells on graft-induced retinal gliosis and retinal ganglion cells survival in diabetic mice

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International Journal of Ophthalmology(English edition) 2024年第1期17卷 34-41页

作者： Tong Wang Hai-Chun Li Jin Ma Xi-Ling Yu State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangdong Provincial Key Laboratory of Ophthalmology and Visual ScienceGuangzhou 510060Guangdong ProvinceChina

AIM:To investigate the effects of Sonic hedgehog(Shh)gene-modified bone marrow mesenchymal stem cells(MSCs)on graft-induced retinal gliosis and retinal ganglion cells(RGCs)survival in diabetic mice.METHODS:Bone marrow-derived MSCs were genetically modified with the Shh gene to generate a stably transfected cell line of Shh-modified MSCs(MSC-Shh).Intravitreal injections of MSC-Shh and green fluorescent protein-modified MSCs(MSC-Gfp;control)were administered in diabetic mice.After 4wk,the effects of MSC-Shh on retinal gliosis were evaluated using fundus photography,and markers of gliosis were examined by immunofluorescence and Western blotting.The neurotrophic factors expression and RGCs survival in the host retina were evaluated using Western blotting and immunofluorescence.The mechanisms underlying the effects of MSC-Shh was investigated.RESULTS:A significant reduction of proliferative vitreoretinopathy(PVR)was observed after intravitreal injection of MSC-Shh compared to MSC-Gfp.Significant downregulation of glial fibrillary acidic protein(GFAP)was demonstrated in the host retina after MSC-Shh administration compared to MSC-Gfp.The extracellular signal-regulated kinase 1/2(ERK1/2),protein kinase B(AKT)and phosphatidylin-ositol-3-kinase(PI3K)pathways were significantly downregulated after MSC-Shh administration compared to MSC-Gfp.Brain-derived neurotrophic factor(BDNF)and ciliary neurotrophic factor(CNTF)levels were significantly increased in the host retina,and RGCs loss was significantly prevented after MSC-Shh administration.CONCLUSION:MSC-Shh administration reduces graft-induced reactive gliosis following intravitreal injection in diabetic mice.The ERK1/2,AKT and PI3K pathways are involved in this process.MSC-Shh also increases the levels of neurotrophic factors in the host retina and promoted RGCs survival in diabetic mice.

关键词： mesenchymal stem cells Sonic hedgehog signaling reactive gliosis diabetic retinopathy retinal ganglion cells

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Neuroprotective effects of resveratrol on retinal ganglion cells in glaucoma in rodents:A narrative review

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Animal Models and Experimental Medicine 2024年第3期7卷 195-207页

作者： Maryam Golmohammadi Seyed Arash Aghaei Meibodi Sulieman Ibraheem Shelash Al-Hawary Jitendra Gupta Ibrohim B.Sapaev Mazin A.A.Najm Marim Alwave Mozhgan Nazifi Mohammadreza Rahmani Mohammad Yasin Zamanian Gervason Moriasi School of Medicine Shahid Beheshti University of Medical SciencesTehranIran Department of Business Administration Business SchoolAl al-Bayt UniversityMafraqJordan Institute of Pharmaceutical Research GLA UniversityMathuraIndia Tashkent Institute of Irrigation and Agricultural Mechanization Engineers TashkentUzbekistan New Uzbekistan University TashkentUzbekistan Pharmaceutical Chemistry Department College of PharmacyAl-Ayen UniversityThi-QarIraq Medical Technical College Al-Farahidi UniversityBaghdadIraq Department of Neurology Hamadan University of Medical SciencesHamadanIran Physiology-Pharmacology Research Center Research Institute of Basic Medical SciencesRafsanjan University of Medical SciencesRafsanjanIran Department of Physiology and Pharmacology School of MedicineRafsanjan University of Medical SciencesRafsanjanIran Neurophysiology Research Center Hamadan University of Medical SciencesHamadanIran Department of Pharmacology and Toxicology School of PharmacyHamadan University of Medical SciencesHamadanIran Department of Medical Biochemistry School of MedicineCollege of Health SciencesMount Kenya UniversityThikaKenya

Glaucoma,an irreversible optic neuropathy,primarily affects retinal ganglion cells(RGC)and causes vision loss and blindness.The damage to RGCs in glaucoma occurs by various mechanisms,including elevated intraocular pressure,oxidative stress,inflammation,and other neurodegenerative processes.As the disease progresses,the loss of RGCs leads to vision loss.Therefore,protecting RGCs from damage and promoting their survival are important goals in managing glaucoma.In this regard,resveratrol(RES),a polyphenolic phytoalexin,exerts antioxidant effects and slows down the evolution and progression of glaucoma.The present review shows that RES plays a protective role in RGCs in cases of ischemic injury and hypoxia as well as in ErbB2 protein expression in the retina.Additionally,RES plays protective roles in RGCs by promoting cell growth,reducing apoptosis,and decreasing oxidative stress in H_(2)O_(2)-exposed RGCs.RES was also found to inhibit oxidative stress damage in RGCs and suppress the activation of mitogen-activated protein kinase signaling pathways.RES could alleviate retinal function impairment by suppressing the hypoxia-i nducible factor-1 alpha/vascular endothelial growth factor and p38/p53 axes while stimulating the PI3K/Akt pathway.Therefore,RES might exert potential therapeutic effects for managing glaucoma by protecting RGCs from damage and promoting their survival.

关键词： glaucoma ischemic-reperfusion injury oxidative stress resveratrol retinal ganglion cells

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A network pharmacology-based strategy for predicting the protect-ive mechanism of Ginkgo biloba on damaged retinal ganglion cells

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Chinese Journal of Natural Medicines 2022年第1期20卷 54-66页

作者： YU Hua DONG Li-Hong ZHANG Yan LIU Qiang Chaohu Hospital Affiliated to Anhui Medical University Hefei 230000China

Hallmarks of the pathophysiology of glaucoma are oxidative stress and apoptotic death of retinal ganglion cells(RGCs).Ginkgo biloba extract(EGb)with multi-target,multi-pathway functions has been reported to exert positive pharmacological effects on oxidative stress and damaged RGCs.However,the ingredients and anti-apoptotic targets of EGb in the treatment of open-angle glaucoma(OAG)have not been fully elucidated.Therefore,in-depth analysis is necessary for further research.Ginkgo biloba-re-lated and anti-apoptotic targets were identified and then combined to obtain the intersection,representing the potential anti-apoptotic targets of Ginkgo biloba.In addition,compound-anti-apoptotic target and OAG-target protein-protein interaction network were merged to obtain five core genes and compound-OAG-anti-apoptotic target protein-protein interaction network.Consequently,the active com-pounds and anti-apoptotic targets of Ginkgo biloba in the treatment of OAG were identified,namely luteolin,β-sitosterol,kaempferol,stigmasterol,quercetin,and p53,Bax,Bcl-2,Caspase-3 and Caspase-9,respectively.For the anti-apoptotic targets of Ginkgo biloba in the treatment of OAG,Gene Ontology(GO)and pathway analysis were executed to confirm the gene functions of Ginkgo biloba in antagonizing apoptosis of RGCs.The pathway enrichment was mainly involved in transcriptional activation of p53 responsive genes,activation of caspases and apoptotic processes.Finally,we confirmed the results of the network analysis by H2O2 treated RGC-5 cells in vitro.The results demonstrated that EGb protection can effectively diminish H2O2-induced apoptosis by inhibiting p53 acetylation,reducing the ratio of Bax/Bcl-2 and suppressing the expression of specific cleavage of Caspase-9 and Caspase-3.

关键词： Ginkgo biloba Oxidative stress retinal ganglion cells Apoptosis p53 Acetylation

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Signaling Mechanism for Modulation by GLP-1 and Exendin-4 of GABA Receptors on Rat retinal ganglion cells

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Neuroscience Bulletin 2022年第6期38卷 622-636页

作者： Tao Zhang Hang-Ze Ruan Yong-Chen Wang Yu-Qi Shao Wei Zhou Shi-Jun Weng Yong-Mei Zhong State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science Institutes of Brain ScienceFudan UniversityShanghai 200032.China

Glucagon-like peptide-1(GLP-1)is expressed in retinal neurons,but its role in the retina is largely unknown.Here,we demonstrated that GLP-1 or the GLP-1 receptor(GLP-1R;a G protein-coupled receptor)agonist exendin-4 suppressed-aminobutyric acid receptor(GABAR)-mediated currents through GLP-1Rs in isolated rat retinal ganglion cells(GCs).Pre-incubation with the stimulatory G protein(G,)inhibitor NF 449 abolished the exendin-4 effect.The exendin-4-induced suppression was mimicked by perfusion with 8-Br-cAMP(a cAMP analog),but was eliminated by the protein kinase A(PKA)inhibitor RpcAMP/KT-5720.The_exendin-4 effect was accompanied by an increase in[Ca2+];of GCs through the IP3-sensitive pathway and was blocked in Ca2+-free solution.Furthermore,when the activity of calmodulin(CaM)and CaMdependent protein kinase II(CaMKII)was inhibited,the exendin-4 effect was eliminated.Consistent with this,exendin-4 suppressed GABAR-mediated light-evoked inhibitory postsynaptic currents in GCs in rat retinal slices.These results suggest that exendin-4-induced suppression may be mediated by a distinct G,/cAMP-PKA/IPs/Ca+/CaM/CaMKIl signaling pathway,following the activation of GLP-1Rs.

关键词： Glucagon-like peptide-1 Exendin-4 GABA current retinal ganglion cells Neuromodulation

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PLSCR1 promotes apoptosis and clearance of retinal ganglion cells in glaucoma pathogenesis

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Genes & Diseases 2023年第4期10卷 1564-1581页

作者： Jingyi Luo Qing Lian Deliang Zhu Minglei Zhao Tingfang Mei Bizhi Shang Zeqiu Yang Chujun Liu Wenchang Xu Lan Zhou Keling Wu Xinqi Liu Yuhua Lai Fuxiang Mao Weihua Li Chengguo Zuo Kang Zhang Mingkai Lin Yehong Zhuo Yizhi Liu Lin Lu Ling Zhao State Key Laboratory of Ophthalmology Zhongshan Ophthalmic CenterSun Yat-sen UniversityGuangdong Provincial Key Laboratory of Ophthalmology and Visual ScienceGuangzhouGuangdong 510060China Dongguan Guangming Ophthalmic Hospital Dongguan523000China Guangdong Provincial People’s Hospital Guangdong Academy of Medical SciencesGuangzhouGuangdong 510080China Department of Ophthalmology Huizhou Municipal Central HospitalHuizhouGuangdong 516000China Guangdong Province Key Laboratory of Brain Function and Disease Zhongshan School of MedicineSun Yat-sen UniversityGuangzhouGuangdong 510000China Center for Biomedicine and Innovations Faculty of MedicineMacao University of Science and TechnologyMacao 999078China Guangzhou Regenerative Medicine and Health Guangdong Laboratory GuangzhouGuangdong 510530China Research Unit of Ocular Development and Regeneration Chinese Academy of Medical SciencesGuangzhouGuangdong 510060China

Glaucoma is the leading cause of irreversible blindness worldwide.In the pathogen-esis of glaucoma,activated microglia can lead to retinal ganglion cells(RGCs)apoptosis and death,however,the molecular mechanisms remain largely unknown.We demonstrate that phospholipid scramblase 1(PLSCR1)is a key regulator promoting RGCs apoptosis and their clearance by microglia.As evidenced in retinal progenitor cells and RGCs of the acute ocular hypertension(AOH)mouse model,overexpressed PLSCR1 induced its translocation from the nucleus to the cytoplasm and cytomembrane,as well as elevated phosphatidylserine exposure and reactive oxygen species generation with subsequent RGCs apoptosis and death.These damages were effectively attenuated by PLSCR1 inhibition.In the AOH model,PLSCR1 led to an increase in M1 type microglia activation and retinal neuroinflammation.Upregulation of PLSCR1 resulted in strongly elevated phagocytosis of apoptotic RGCs by activated microglia.Taken together,our study provides important insights linking activated microglia to RGCs death in the glaucoma pathogenesis and other RGC-related neurodegenerative diseases.

关键词： Apoptosis Glaucoma Phagocytosis PLSCR1 retinal ganglion cells

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Influence of hypoxia on retinal progenitor and ganglion cells in human induced pluripotent stem cell-derived retinal organoids

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国际眼科杂志:英文版 2023年第10期16卷 1574-1581页

作者： Jin-Lin Du Li-Xiong Gao Tao Wang Zi Ye Hong-Yu Li Wen Li Quan Zeng Jia-Fei Xi Wen Yue Zhao-Hui Li Medical School of Chinese PLA Beijing 100853China Senior Department of Ophthalmology the Third Medical Center of Chinese PLA General HospitalBeijing 100039China Departement of Ophthalmology the 6^(th)Medical Center of PLA General HospitalBeijing 100048China Stem Cell and Regenerative Medicine Lab Beijing Institute of Radiation MedicineBeijing 100850China

AIM:To observe the effect of low oxygen concentration on the neural retina in human induced pluripotent stem cell(hiPSC)-derived retinal organoids(ROs).METHODS:The hiPSC and a three-dimensional culture method were used for the experiments.Generated embryoid bodies(EBs)were randomly and equally divided into hypoxic and normoxic groups.Photographs of the EBs were taken on days 38,45,and 52,and the corresponding volume of EBs was calculated.Simultaneously,samples were collected at these three timepoints,followed by fixation,sectioning,and immunofluorescence.RESULTS:The proportion of Ki67-positive proliferating cells increased steadily on day 38;this proliferationpromoting effect tended to increase tissue density rather than tissue volume.On days 45 and 52,the two groups had relatively similar ratios of Ki67-positive cells.Further immunofluorescence analysis showed that the ratio of SOX2-positive cells significantly increased within the neural retina on day 52(Pcells significantly decreased following hypoxia treatment at all three timepoints(P0.05).Moreover,the proportion of PAX6-/TUJ1+cells within the neural retinas increased considerably(Pretinal progenitor cell pool in hiPSC-derived ROs.

关键词： hypoxia retinal organoid retinal progenitor cells retinal ganglion cells

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High glucose levels impact visual response properties of retinal ganglion cells in C57 mice—An in vitro physiological study

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Science China(Life Sciences) 2017年第12期60卷 1428-1435页

作者： Yuan Zhou Chunxia Xiao Mingliang Pu Department of Anatomy/Embryology School of Basic Medical Sciences Peking University Functional Department Mawangdui Branch People's Hospital of Hunan Province The First Affiliated Hospital of Hunan Normal University

This study investigated visual response properties of retinal ganglion cells(RGCs) under high glucose levels. Extracellular single-unit responses of RGCs from mouse retinas were recorded. And the eyecup was prepared as a flat mount in a recording chamber and superfused with Ames medium. The averaged RF size of the ON RGCs(34.1±2.9, n=14) was significantly smaller than the OFF RGCs under the HG(49.3±0.3, n=12)(Pcells was significantly lower than that of OFF cells with the HM treatment(Pcells was significantly higher than OFF cells with the HG treatment(P<0.0001). The present results suggest that HG reduced receptive field center size, suppressed luminance threshold, and attenuated contrast gain of RGCs. The impact of HG on ON and OFF RGCs may be mediated via different mechanisms.

关键词： retinal ganglion cells high glucose high mannitol receptive field center size luminance threshold contrast threshold

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Dose-dependent and combined effects of N-methyl-D-aspartate receptor antagonist MK-801 and nitric oxide synthase inhibitor nitro-L-arginine on the survival of retinal ganglion cells in adult hamsters

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Neural Regeneration Research 2012年第10期7卷 725-730页

作者： Yaoyu Li An＇an Yang Tingting Zhu Zhao Liu Siwei You Kwok-Fai So Department of Ophthalmology General Hospital of Beijing Military Region Beijing 100700 China Institute of Neurosciences the Fourth Military Medical University Xi＇an 710032 Shaanxi Province China Department of Anatomy Li Ka Shing Faculty of Medicine The University of Hong Kong Hong Kong Special Administrative Region China

This study investigated the effects of daily intraperitoneal injections of N-methyl-D-aspartate receptor antagonist MK-801 and nitric oxide synthase inhibitor nitro-L-arginine （L-NA） on the survival of retinal ganglion cells （RGCs） at 1 and 2 weeks after unilateral optic nerve transection in adult hamsters. The left optic nerves of all animals were transected intraorbitally 1 mm from the optic disc and RGCs were retrogradely labeled with Fluorogold before they received different daily dosages of single MK-801 or L-NA as well as daily combinational treatments of these two chemicals. All experimental and control animals survived for 1 or 2 weeks after optic nerve transection. Our results revealed that the mean numbers of surviving RGCs increased and then decreased when the dosage of MK-801 （1.0, 3.0 and 4.5 mg/kg） and L-NA （1.5, 3.0, 4.5 and 6.0 mg/kg） increased at both 1 and 2 weeks survival time points. Daily combinational use of 1.0 mg/kg MK-801 and 1.5 mg/kg L-NA lead to a highest RGC number that was even higher than the sum of the RGC numbers in 1.0 mg/kg MK-801 and 1.5 mg/kg L-NA subgroups at 2 weeks. These findings indicated that both MK-801 and L-NA can protect axotomized RGCs in a dose-dependent manner and combinational treatment of these chemicals possesses a potentiative and protective effect.

关键词： retinal ganglion cells optic nerve transection N-methyl-D-aspartate receptor nitric oxide synthase neuronal survival hamster

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Minocycline protects retinal ganglion cells after optic nerve crush injury in mice by delaying autophagy and upregulating nuclear factor-κB2

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Chinese Medical Journal 2014年第9期127卷 1749-1754页

作者： Jiao Xiaoling Peng Yuan Yang Liu Department of Ophthalmology Peking University First Hospital Key Laboratory of Vision Loss and Restoration Ministry of Education Beijing 100034 China

Background Currently,no medicine is available that can prevent or treat neural damage associated with optic nerve injury.Minocycline is recently reported to have a neuroprotective function.The aims of this study were to exarmine the neuroprotective effect of minocycline on retinal ganglion cells （RGCs） and determine its underlying mechanisms,using a mouse model of optic nerve crush （ONC）.Methods ONC was performed in the left eye of adult male mice,and the mice were randomly divided into minocycline-treated group and saline-treated control group.The mice without receiving ONC injury were used as positive controls.RGC densities were assessed in retinal whole mounts with immunofluorescence labeling of βⅢ-tubulin.Transmission electron microscopy was used to detect RGC morphologies,and Western blotting and real-time PCR were applied to investigate the expression of autophagy markers LC3-Ⅰ,LC3-Ⅱ,and transcriptional factors nuclear factor-κB1 （NF-κB1）,NF-κB2.Results In the early stage after ONC （at Days 4 and 7）,the density of RGCs in the minocycline-treated group was higher than that of the saline-treated group.Electron micrographs showed that minocycline prevented nuclei and mitochondria injuries at Day 4.Western blotting analysis demonstrated that the conversion of LC3-Ⅰ to LC3-Ⅱ was reduced in the minocycline-treated group at Days 4 and 7,which meant autophagy process was inhibited by minocycline.In addition,the gene expression of NF-κB2 was upregulated by minocycline at Day 4.Conclusion The neuroprotective effect of minocycline is generated in the early stage after ONC in mice,partly through delaying autophagy process and regulating NF-κB2 pathway.

关键词： minocycline autophagy LC3 protein nuclearfactor-κB2 p52 subunit retinal ganglion cells optic nerve injury

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Cell transplantation to replace retinal ganglion cells faces challenges-the Switchboard Dilemma

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Neural Regeneration Research 2021年第6期16卷 1138-1143页

作者： Yuan Liu Richard K.Lee Bascom Palmer Eye Institute University of Miami Miller School of MedicineMiamiFLUSA

The mammalian retina displays incomplete intrinsic regenerative capacities;therefore,retina degeneration is a major cause of irreversible blindness such as glaucoma,agerelated macular degeneration and diabetic retinopathy.These diseases lead to the loss of retinal cells and serious vision loss in the late stage.Stem cell transplantation is a great promising novel treatment for these incurable retinal degenerative diseases and represents an exciting area of regenerative neurotherapy.Several suitable stem cell sources for transplantation including human embryonic stem cells,induced pluripotent stem cells and adult stem cells have been identified as promising target populations.However,the retina is an elegant neuronal complex composed of various types of cells with different functions.The replacement of these different types of cells by transplantation should be addressed separately.So far,retinal pigment epithelium transplantation has achieved the most advanced stage of clinical trials,while transplantation of retinal neurons such as retinal ganglion cells and photoreceptors has been mostly studied in pre-clinical animal models.In this review,we opine on the key problems that need to be addressed before stem cells transplantation,especially for replacing injured retinal ganglion cells,may be used practically for treatment.A key problem we have called the Switchboard Dilemma is a major block to have functional retinal ganglion cell replacement.We use the public switchboard telephone network as an example to illustrate different difficulties for replacing damaged components in the retina that allow for visual signaling.retinal ganglion cell transplantation is confronted by significant hurdles,because retinal ganglion cells receive signals from different interneurons,integrate and send signals to the correct targets of the visual system,which functions similar to the switchboard in a telephone network-therefore the Switchboard Dilemma.

关键词： cell transplantation optic nerve regeneration photoreceptors retina degeneration retinal ganglion cells stem cells

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