BACKGROUND Diabetic retinopathy(DR)is one of the major eye diseases contributing to blindness worldwide.Endoplasmic reticulum(ER)stress in retinal cells is a key factor leading to retinal inflammation and vascular lea...BACKGROUND Diabetic retinopathy(DR)is one of the major eye diseases contributing to blindness worldwide.Endoplasmic reticulum(ER)stress in retinal cells is a key factor leading to retinal inflammation and vascular leakage in DR,but its mechanism is still unclear.AIM To investigate the potential mechanism of LEF1 and related RNAs in DR.METHODS ARPE-19 cells were exposed to high levels of glucose for 24 hours to simulate a diabetic environment.Intraperitoneally injected streptozotocin was used to induce the rat model of DR.The expression levels of genes and related proteins were measured by RT-qPCR and Western blotting;lnc-MGC and miR-495-3p were detected by fluorescent in situ hybridization;CCK-8 and TUNEL assays were used to detect cell viability and apoptosis;enzyme-linked immunosorbent assay was used to detect inflammatory factors;dual-luciferase gene assays were used to verify the targeting relationship;and the retina was observed by HE staining.RESULTS LEF1 and lnc-MGC have binding sites,and lnc-MGC can regulate the miR-495-3p/GRP78 molecular axis.In high glucose-treated cells,inflammation was aggravated,the intracellular reactive oxygen species concentration was increased,cell viability was reduced,apoptosis was increased,the ER response was intensified,and ferroptosis was increased.As an ER molecular chaperone,GRP78 regulates the ER and ferroptosis under the targeting of miR-495-3p,whereas inhibiting LEF1 can further downregulate the expression of lnc-MGC,increase the level of miR-495-3p,and sequentially regulate the level of GRP78 to alleviate the occurrence and development of DR.Animal experiments indicated that the knockdown of LEF1 can affect the lnc-MGC/miR-495-3p/GRP78 signaling axis to restrain the progression of DR.CONCLUSION LEF1 knockdown can regulate the miR-495-3p/GRP78 molecular axis through lnc-MGC,which affects ER stress and restrains the progression of DR and ferroptosis in retinal pigment epithelial cells.展开更多
Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78（DRi P78） and Na＋-H＋ exchanger r...Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78（DRi P78） and Na＋-H＋ exchanger regulatory factor 1（NHERF1） are the CXCR4 and CCR5 homo- or hetero-dimerinteracting proteins. DRi P78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRi P78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs（sh RNAs） targeting the DRi P78 and NHERF1, respectively, and constructed the p Lenti6/BLOCK-i T-DEST lentiviral plasmids expressing DRi P78 or NHERF1 sh RNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST（3）. Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST（3） with DRi P78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.展开更多
We identified a leafy head mutant plal-5 （plastochron 1-5） from the progeny of japonica rice cultivar Taipei 309 treated with 60Co-γ ray irradiation. The plal-5 mutant has a dwarf phenotype and small leaves. Compar...We identified a leafy head mutant plal-5 （plastochron 1-5） from the progeny of japonica rice cultivar Taipei 309 treated with 60Co-γ ray irradiation. The plal-5 mutant has a dwarf phenotype and small leaves. Compared with its wild type, plal-5 has more leaves and fewer tillers, and it fails to produce normal panicles at the maturity stage. Genetic analysis showed that the plal-5 phenotype is controlled by a single recessive nuclear gene. Using the map-based cloning strategy, we narrowed down the location of the target gene to a 58-kb region between simple sequence repeat markers CHR1027 and CHR1030 on the long arm of chromosome 10. The target gene cosegregated with molecular markers CHR1028 and CHR1029. There were five predicted genes in the mapped region. The results from sequencing analysis revealed that there was one base deletion in the first exon of LOC_Os10g26340 encoding cytochrome P450 CYP78A11 in the plal-5 mutant, which might result in a downstream frame shift and premature termination. These results suggest that the P450 CYP78A11 gene is the candidate gene of PLA1-5.展开更多
基金Supported by Science and Technology Program of Yunnan Provincial Department of Science and Technology-Basic Research Program,No.202301BA070001-025.
文摘BACKGROUND Diabetic retinopathy(DR)is one of the major eye diseases contributing to blindness worldwide.Endoplasmic reticulum(ER)stress in retinal cells is a key factor leading to retinal inflammation and vascular leakage in DR,but its mechanism is still unclear.AIM To investigate the potential mechanism of LEF1 and related RNAs in DR.METHODS ARPE-19 cells were exposed to high levels of glucose for 24 hours to simulate a diabetic environment.Intraperitoneally injected streptozotocin was used to induce the rat model of DR.The expression levels of genes and related proteins were measured by RT-qPCR and Western blotting;lnc-MGC and miR-495-3p were detected by fluorescent in situ hybridization;CCK-8 and TUNEL assays were used to detect cell viability and apoptosis;enzyme-linked immunosorbent assay was used to detect inflammatory factors;dual-luciferase gene assays were used to verify the targeting relationship;and the retina was observed by HE staining.RESULTS LEF1 and lnc-MGC have binding sites,and lnc-MGC can regulate the miR-495-3p/GRP78 molecular axis.In high glucose-treated cells,inflammation was aggravated,the intracellular reactive oxygen species concentration was increased,cell viability was reduced,apoptosis was increased,the ER response was intensified,and ferroptosis was increased.As an ER molecular chaperone,GRP78 regulates the ER and ferroptosis under the targeting of miR-495-3p,whereas inhibiting LEF1 can further downregulate the expression of lnc-MGC,increase the level of miR-495-3p,and sequentially regulate the level of GRP78 to alleviate the occurrence and development of DR.Animal experiments indicated that the knockdown of LEF1 can affect the lnc-MGC/miR-495-3p/GRP78 signaling axis to restrain the progression of DR.CONCLUSION LEF1 knockdown can regulate the miR-495-3p/GRP78 molecular axis through lnc-MGC,which affects ER stress and restrains the progression of DR and ferroptosis in retinal pigment epithelial cells.
基金supported by the Science and Technology Planning Project of Guangdong Province of China（2012B031800267）the Natural Science Foundation of Guangdong Province of China（S2013010011860）the National Natural Science Foundation of China（31200130 and 81371812）
文摘Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78（DRi P78） and Na＋-H＋ exchanger regulatory factor 1（NHERF1） are the CXCR4 and CCR5 homo- or hetero-dimerinteracting proteins. DRi P78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRi P78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs（sh RNAs） targeting the DRi P78 and NHERF1, respectively, and constructed the p Lenti6/BLOCK-i T-DEST lentiviral plasmids expressing DRi P78 or NHERF1 sh RNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST（3）. Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST（3） with DRi P78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.
基金financially supported by grants from the Distinguished Young Scientists from Jiangsu GovernmentChina(Grant No.BK2012010)+1 种基金the Key Project of Chinese Ministry of Educationand the Ministry of Science and Technology of China(Grant Nos.2012AA10A302-7 and 2013ZX08009-003)
文摘We identified a leafy head mutant plal-5 （plastochron 1-5） from the progeny of japonica rice cultivar Taipei 309 treated with 60Co-γ ray irradiation. The plal-5 mutant has a dwarf phenotype and small leaves. Compared with its wild type, plal-5 has more leaves and fewer tillers, and it fails to produce normal panicles at the maturity stage. Genetic analysis showed that the plal-5 phenotype is controlled by a single recessive nuclear gene. Using the map-based cloning strategy, we narrowed down the location of the target gene to a 58-kb region between simple sequence repeat markers CHR1027 and CHR1030 on the long arm of chromosome 10. The target gene cosegregated with molecular markers CHR1028 and CHR1029. There were five predicted genes in the mapped region. The results from sequencing analysis revealed that there was one base deletion in the first exon of LOC_Os10g26340 encoding cytochrome P450 CYP78A11 in the plal-5 mutant, which might result in a downstream frame shift and premature termination. These results suggest that the P450 CYP78A11 gene is the candidate gene of PLA1-5.
