AIM: To assess the efficacy of topical Semaphorin 3A(SEMA3A) in the treatment of allergic conjunctivitis.METHODS: Experimental allergic conjunctivitis(EAC)mice model induced by short ragweed pollen(SRW) in 4-week-old ...AIM: To assess the efficacy of topical Semaphorin 3A(SEMA3A) in the treatment of allergic conjunctivitis.METHODS: Experimental allergic conjunctivitis(EAC)mice model induced by short ragweed pollen(SRW) in 4-week-old of BALB/c mice, mice were evaluated using haematoxylin and eosin(H&E) staining, immunofluor-escence and light microscope photographs. Early phase took the samples in 24 h after instillation and late phase took the samples between 4 to 14 d after the start of treatment. The study use of topical SEMA3A(10 U, 100 U,1000 U) eye drops and subconjunctival injection of SEMA3 A with same concentration. For comparison, five types of allergy eyedrops were quantified using clinical characteristics.· RESULTS: Clinical score of composite ocular symptoms of the mice treated with SEMA3 A were significantly decreased both in the immediate phase and the late phase compared to those treated with commercial ophthalmic formulations and non-treatment mice. SEMA3 A treatment attenuates infiltration of eosinophils entering into conjunctiva in EAC mice. The score of eosinophil infiltration in the conjunctiva of SEMA3 A 1000 U-treated group were significantly lower than low-concentration of SEMA3 A treated groups and non-treated group. SEMA3 A treatment also suppressed T-cell proliferation in vitro and decreased serum total Ig E levels in EAC mice. Moreover, treatment of SEMA3 A suppressed Th2-related cytokines(IL-5, IL-13 and IL-4)and pro-inflammatory cytokines(IFN-γ, IL-17 and TNF-α)release, but increased regulatory cytokine IL-10 concentration in the conjunctiva of EAC mice.CONCLUSION: SEMA3 A as a biological agent, showed the beneficial activity in ocular allergic processes with the less damage to the intraocular tissue. It is expected that SEMA3 A may be contributed in patients with a more severe spectrum of refractory ocular allergic diseases including allergic conjunctivitis in the near future.展开更多
AIM： To elucidate the role of neuropilin-1 （Nrp-1） and semaphorin 3A （Sema3A） in sinusoidal remodeling dur- ing liver regeneration in rats. METHODS： Male Wistar/ST rats at 7 wk of age, weigh- ing about 200 g, we...AIM： To elucidate the role of neuropilin-1 （Nrp-1） and semaphorin 3A （Sema3A） in sinusoidal remodeling dur- ing liver regeneration in rats. METHODS： Male Wistar/ST rats at 7 wk of age, weigh- ing about 200 g, were used for all animal experiments. In vivo, at 24, 48, 72, 96, 144 and 192 h after two- thirds partial hepatectomy （PHx）, the remnant livers were removed. Liver tissues were immunohistochemi- cally stained for Nrp-1, Sema3A and SE-1, a liver sinu- soidal endothelial cell （SEC） marker. Total RNA of the liver tissue was extracted and reversely transcribed into cDNA. The mRNA expression of Sema3A was ana- lyzed by quantitative real-time polymerase chain reac- tion and normalized to that of ribosomal protein $18. In vitro, SECs were isolated from rat liver and cultured in endothelial growth medium containing 20 ng/mL vascular endothelial cell growth factor. Migration of SECs in primary culture was assessed by cell transwell assay with or without recombinant Sema3A. Apoptotic cells were determined by a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method. RESULTS： In vitro, immunohistochemistry study re- vealed that Sema3A and Nrp-1 were constitutively ex- pressed in hepatocytes and SECs, respectively, in normal rat liver tissues. Nrp-1 expression in SECs was quantified by the percentage of immunostained area with anti- Nrp-1 antibody in relation to the area stained with SE-1. Between 24 h and 96 h following resection of liver, Nrp-1 expression in SECs was transiently increased. Compared with the baseline （5.2% ± 0.1%）, Nrp-1 expression in SECs significantly increased at 24 h （17.3% ± 0.7%, P 〈 0.05）, 48 h （39.1% ± 0.6%, P 〈 0.01）, 72 h （46.9% ± 4.5%, P 〈 0.01） and 96 h （29.9% ±3.8%, P 〈 0.01） after PHx, then returned to the basal level at termination of liver regeneration. Interestingly, the expression of Sema3A was inversely associated with that of Nrp-1 in liver after PHx. Sema3A mRNA expres- sion was significantly reduced by about 75% over the period 24-144 h after PHx （P 〈 0.05）, and returned to basal levels at 192 h after PHx. In vitro, SECs isolated from rats after PHx （PHx-SECs） were observed to mi- grate to the lower chamber of the cell transwell system after incubation for 24 h, but not cells from normal rats （CONT-SECs）, indicating that mobility of PHx-SECs increases as compared with that of CONT-SECs. More- over, recombinant Sema3A significantly attenuated mi- gration in PHx-SECs in primary culture （vehicle-treated 100% ± 7.9% vs Sema3A-treated 42.6% ± 5.4%, P 〈 0.01）, but not in CONT-SECs. Compared with CONT- SECs, the apoptotic rate of PHx-SECs decreased by 78.3% （P 〈 0.05）. There was no difference in apopto- sis between CONT-SECs that were treated with vehicle and Sema3A. However, in PHx-SECs, apoptosis was induced by the presence of 5 nmol Sema3A for 24 h （vehicle-treated 21.7%±7.6% vs Sema3A-treated 104.3% ± 8.9%, P 〈 0.05）. In addition, immunohisto- chemistry confirmed the increased expression of Nrp-1 in PHx-SECs, while it was noted to a lesser extent in CONT-SECs. CONCLUSION： The interplay of Nrp-1 and Sema3A shown in our results may lead to a better understand- ing of interaction between sinusoidal remodeling and SECs during liver regeneration.展开更多
Highlight Semaphorin 3A is a classically known axonal guidance cue that mediates axonal growth cone repulsion and collapse.Recent works,however,suggest that it may have the apparently diametrically opposite activity o...Highlight Semaphorin 3A is a classically known axonal guidance cue that mediates axonal growth cone repulsion and collapse.Recent works,however,suggest that it may have the apparently diametrically opposite activity of promoting neuronal regeneration.展开更多
The expression of nerve cell adhesion molecule L1 in the neuronal growth cone of the central nervous system is strongly associated with the direction of growth of the axon, but its role in the regeneration of the peri...The expression of nerve cell adhesion molecule L1 in the neuronal growth cone of the central nervous system is strongly associated with the direction of growth of the axon, but its role in the regeneration of the peripheral nerve is still unknown. This study explored the problem in a femoral nerve section model in rats. L1 and semaphorin 3A m RNA and protein expressions were measured over the 4-week recovery period. Quantitative polymerase chain reaction showed that nerve cell adhesion molecule L1 expression was higher in the sensory nerves than in motor nerves at 2 weeks after injury, but vice versa for the expression of semaphorin 3A. Western blot assay results demonstrated that nerve cell adhesion molecule L1 expression was higher in motor nerves than in the sensory nerves at the proximal end after injury, but its expression was greater in the sensory nerves at 2 weeks. Semaphorin 3A expression was higher in the motor nerves than in the sensory nerves at 3 days and 1 week after injury. Nerve cell adhesion molecule L1 and semaphorin 3A expressions at the distal end were higher in the motor nerves than in the sensory nerves at 3 days, 1 and 2 weeks. Immunohistochemical staining results showed that nerve cell adhesion molecule L1 expression at the proximal end was greater in the sensory nerves than in the motor nerves; semaphorin 3A expression was higher in the motor nerves than in the sensory nerves at 2 weeks after injury. Taken together, these results indicated that nerve cell adhesion molecules L1 and semaphorin 3A exhibited different expression patterns at the proximal and distal ends of sensory and motor nerves, and play a coordinating role in neural chemotaxis regeneration.展开更多
Semaphorin 3A expression is thought to increase following spinal cord injury.The impact of olfactory ensheathing cell transplantation remains unclear.The current study demonstrated that spinal cord hemorrhage,edema,de...Semaphorin 3A expression is thought to increase following spinal cord injury.The impact of olfactory ensheathing cell transplantation remains unclear.The current study demonstrated that spinal cord hemorrhage,edema,degeneration,necrosis,cyst formation,proliferation of glial cells,regeneration of nerve fibers and various pathological reactions occurred following a simple cross-section of spinal cord injury.Transplantation of olfactory ensheathing cells was found to significantly relieve the pathological reactions in the spinal cord described above,decrease the extent of necrosis in damaged neurons and nerve fibers,and downregulate semaphorin 3A expression in the injured zone.The results confirmed that olfactory ensheathing cell transplantation plays a protective role on the injured spinal cord by reducing the expression of semaphorin 3A.展开更多
Semaphorin 3A could be involved in angiogenesis and also enhanced bone formation was investigated in many researches. In our current study, we firstly investigated that canal obliteration might be present in some rege...Semaphorin 3A could be involved in angiogenesis and also enhanced bone formation was investigated in many researches. In our current study, we firstly investigated that canal obliteration might be present in some regenerative endodontic procedures cases clinically. After the establishment of the model of apical periodontitis in the maxillary first molars of rats, pulp revascularization was performed in mesial root canal. Histological sections showed that most of the tissues growing into the root canal were not real pulp tissue, but cementoid, osteoid and periodontal-like membrane. Moreover, we detected that the expression of Semaphorin 3A increased in the mesial root canal. When we used CoCl<sub>2</sub> to induce hypoxic environment, the expression of genes and proteins, Hypoxia inducible factor-1α, Vascular endothelial growth factor, and Semaphorin 3A in dental pulp stem cells were both upregulated. In conclusion, hypoxia mediated the high expression of Semaphorin 3A in DPSC might be involved tissue regeneration during pulp vascularization.展开更多
Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolyti...Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.展开更多
基金A grant-in-aid for Scientific Research from the Japanese Ministry of Education,Culture,Sports,Science and Technology
文摘AIM: To assess the efficacy of topical Semaphorin 3A(SEMA3A) in the treatment of allergic conjunctivitis.METHODS: Experimental allergic conjunctivitis(EAC)mice model induced by short ragweed pollen(SRW) in 4-week-old of BALB/c mice, mice were evaluated using haematoxylin and eosin(H&E) staining, immunofluor-escence and light microscope photographs. Early phase took the samples in 24 h after instillation and late phase took the samples between 4 to 14 d after the start of treatment. The study use of topical SEMA3A(10 U, 100 U,1000 U) eye drops and subconjunctival injection of SEMA3 A with same concentration. For comparison, five types of allergy eyedrops were quantified using clinical characteristics.· RESULTS: Clinical score of composite ocular symptoms of the mice treated with SEMA3 A were significantly decreased both in the immediate phase and the late phase compared to those treated with commercial ophthalmic formulations and non-treatment mice. SEMA3 A treatment attenuates infiltration of eosinophils entering into conjunctiva in EAC mice. The score of eosinophil infiltration in the conjunctiva of SEMA3 A 1000 U-treated group were significantly lower than low-concentration of SEMA3 A treated groups and non-treated group. SEMA3 A treatment also suppressed T-cell proliferation in vitro and decreased serum total Ig E levels in EAC mice. Moreover, treatment of SEMA3 A suppressed Th2-related cytokines(IL-5, IL-13 and IL-4)and pro-inflammatory cytokines(IFN-γ, IL-17 and TNF-α)release, but increased regulatory cytokine IL-10 concentration in the conjunctiva of EAC mice.CONCLUSION: SEMA3 A as a biological agent, showed the beneficial activity in ocular allergic processes with the less damage to the intraocular tissue. It is expected that SEMA3 A may be contributed in patients with a more severe spectrum of refractory ocular allergic diseases including allergic conjunctivitis in the near future.
基金Supported by A Grant-in-aid for Young Scientists from Japan Society for the Promotion of Science,No.22790671
文摘AIM： To elucidate the role of neuropilin-1 （Nrp-1） and semaphorin 3A （Sema3A） in sinusoidal remodeling dur- ing liver regeneration in rats. METHODS： Male Wistar/ST rats at 7 wk of age, weigh- ing about 200 g, were used for all animal experiments. In vivo, at 24, 48, 72, 96, 144 and 192 h after two- thirds partial hepatectomy （PHx）, the remnant livers were removed. Liver tissues were immunohistochemi- cally stained for Nrp-1, Sema3A and SE-1, a liver sinu- soidal endothelial cell （SEC） marker. Total RNA of the liver tissue was extracted and reversely transcribed into cDNA. The mRNA expression of Sema3A was ana- lyzed by quantitative real-time polymerase chain reac- tion and normalized to that of ribosomal protein $18. In vitro, SECs were isolated from rat liver and cultured in endothelial growth medium containing 20 ng/mL vascular endothelial cell growth factor. Migration of SECs in primary culture was assessed by cell transwell assay with or without recombinant Sema3A. Apoptotic cells were determined by a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method. RESULTS： In vitro, immunohistochemistry study re- vealed that Sema3A and Nrp-1 were constitutively ex- pressed in hepatocytes and SECs, respectively, in normal rat liver tissues. Nrp-1 expression in SECs was quantified by the percentage of immunostained area with anti- Nrp-1 antibody in relation to the area stained with SE-1. Between 24 h and 96 h following resection of liver, Nrp-1 expression in SECs was transiently increased. Compared with the baseline （5.2% ± 0.1%）, Nrp-1 expression in SECs significantly increased at 24 h （17.3% ± 0.7%, P 〈 0.05）, 48 h （39.1% ± 0.6%, P 〈 0.01）, 72 h （46.9% ± 4.5%, P 〈 0.01） and 96 h （29.9% ±3.8%, P 〈 0.01） after PHx, then returned to the basal level at termination of liver regeneration. Interestingly, the expression of Sema3A was inversely associated with that of Nrp-1 in liver after PHx. Sema3A mRNA expres- sion was significantly reduced by about 75% over the period 24-144 h after PHx （P 〈 0.05）, and returned to basal levels at 192 h after PHx. In vitro, SECs isolated from rats after PHx （PHx-SECs） were observed to mi- grate to the lower chamber of the cell transwell system after incubation for 24 h, but not cells from normal rats （CONT-SECs）, indicating that mobility of PHx-SECs increases as compared with that of CONT-SECs. More- over, recombinant Sema3A significantly attenuated mi- gration in PHx-SECs in primary culture （vehicle-treated 100% ± 7.9% vs Sema3A-treated 42.6% ± 5.4%, P 〈 0.01）, but not in CONT-SECs. Compared with CONT- SECs, the apoptotic rate of PHx-SECs decreased by 78.3% （P 〈 0.05）. There was no difference in apopto- sis between CONT-SECs that were treated with vehicle and Sema3A. However, in PHx-SECs, apoptosis was induced by the presence of 5 nmol Sema3A for 24 h （vehicle-treated 21.7%±7.6% vs Sema3A-treated 104.3% ± 8.9%, P 〈 0.05）. In addition, immunohisto- chemistry confirmed the increased expression of Nrp-1 in PHx-SECs, while it was noted to a lesser extent in CONT-SECs. CONCLUSION： The interplay of Nrp-1 and Sema3A shown in our results may lead to a better understand- ing of interaction between sinusoidal remodeling and SECs during liver regeneration.
文摘Highlight Semaphorin 3A is a classically known axonal guidance cue that mediates axonal growth cone repulsion and collapse.Recent works,however,suggest that it may have the apparently diametrically opposite activity of promoting neuronal regeneration.
基金supported by the National Natural Science Foundation of China,No.81371389,31500927,31300942,81201017the Collegiate Natural Science Foundation of Jiangsu Province of China,No.13KJB180018the Natural Science Foundation of Nantong University of China,No.14ZY013
文摘The expression of nerve cell adhesion molecule L1 in the neuronal growth cone of the central nervous system is strongly associated with the direction of growth of the axon, but its role in the regeneration of the peripheral nerve is still unknown. This study explored the problem in a femoral nerve section model in rats. L1 and semaphorin 3A m RNA and protein expressions were measured over the 4-week recovery period. Quantitative polymerase chain reaction showed that nerve cell adhesion molecule L1 expression was higher in the sensory nerves than in motor nerves at 2 weeks after injury, but vice versa for the expression of semaphorin 3A. Western blot assay results demonstrated that nerve cell adhesion molecule L1 expression was higher in motor nerves than in the sensory nerves at the proximal end after injury, but its expression was greater in the sensory nerves at 2 weeks. Semaphorin 3A expression was higher in the motor nerves than in the sensory nerves at 3 days and 1 week after injury. Nerve cell adhesion molecule L1 and semaphorin 3A expressions at the distal end were higher in the motor nerves than in the sensory nerves at 3 days, 1 and 2 weeks. Immunohistochemical staining results showed that nerve cell adhesion molecule L1 expression at the proximal end was greater in the sensory nerves than in the motor nerves; semaphorin 3A expression was higher in the motor nerves than in the sensory nerves at 2 weeks after injury. Taken together, these results indicated that nerve cell adhesion molecules L1 and semaphorin 3A exhibited different expression patterns at the proximal and distal ends of sensory and motor nerves, and play a coordinating role in neural chemotaxis regeneration.
文摘Semaphorin 3A expression is thought to increase following spinal cord injury.The impact of olfactory ensheathing cell transplantation remains unclear.The current study demonstrated that spinal cord hemorrhage,edema,degeneration,necrosis,cyst formation,proliferation of glial cells,regeneration of nerve fibers and various pathological reactions occurred following a simple cross-section of spinal cord injury.Transplantation of olfactory ensheathing cells was found to significantly relieve the pathological reactions in the spinal cord described above,decrease the extent of necrosis in damaged neurons and nerve fibers,and downregulate semaphorin 3A expression in the injured zone.The results confirmed that olfactory ensheathing cell transplantation plays a protective role on the injured spinal cord by reducing the expression of semaphorin 3A.
文摘Semaphorin 3A could be involved in angiogenesis and also enhanced bone formation was investigated in many researches. In our current study, we firstly investigated that canal obliteration might be present in some regenerative endodontic procedures cases clinically. After the establishment of the model of apical periodontitis in the maxillary first molars of rats, pulp revascularization was performed in mesial root canal. Histological sections showed that most of the tissues growing into the root canal were not real pulp tissue, but cementoid, osteoid and periodontal-like membrane. Moreover, we detected that the expression of Semaphorin 3A increased in the mesial root canal. When we used CoCl<sub>2</sub> to induce hypoxic environment, the expression of genes and proteins, Hypoxia inducible factor-1α, Vascular endothelial growth factor, and Semaphorin 3A in dental pulp stem cells were both upregulated. In conclusion, hypoxia mediated the high expression of Semaphorin 3A in DPSC might be involved tissue regeneration during pulp vascularization.
基金supported by the National Natural Science Foundation of China(No.81974321)the Discipline Innovation and Talent Introduction Program to Universities from Ministry of Education of China(Project 111)+3 种基金the Tackling Project for Science and Technology of Xinxiang City(No.GG2019003)the Natural Science Foundation of Henan Province of China(No.212300410173)the State Administration of Traditional Chinese Medicine Young Qi Huang Scholar,and the Innovation Team Project of Scientific Research of Traditional Chinese Medicine of Shanghai Health Committee(No.2022CX001)China.
文摘Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.
