Based on the structure of FK506, FKBP12 and calcineurin complex and the interactivecharacteristics of small molecular ligands with FKBPs, a series of L-1,4-thiazane-3-carboxylic acidderivatives as neuroimmunophilin li...Based on the structure of FK506, FKBP12 and calcineurin complex and the interactivecharacteristics of small molecular ligands with FKBPs, a series of L-1,4-thiazane-3-carboxylic acidderivatives as neuroimmunophilin ligands was designed and synthesized. The results of evaluationshow that compound N308 has a great promise as a candidate of neuroprotective andneuroregenerative agent.展开更多
Molting and metamorphosis are important physiological processes in insects that are tightly controlled by ecdysone receptor(EcR)through the 20-hydroxyecdysone(20E)signaling pathway.EcR is a steroid nuclear receptor(SR...Molting and metamorphosis are important physiological processes in insects that are tightly controlled by ecdysone receptor(EcR)through the 20-hydroxyecdysone(20E)signaling pathway.EcR is a steroid nuclear receptor(SR).Several FK506-binding proteins(FKBPs)have been identified from the mammal SR complex,and are thought to be involved in the subcellular trafficking of SR.However,their roles in insects are poorly understood.To explore whether FKBPs are involved in insect molting or metamorphosis,we injected an FKBP inhibitor(FK506)into a lepidopteran insect,Spodoptera litura,and found that molting was inhibited in 61.11%of the larvae,and that the time for larvae to pupate was significantly extended.A total of 10 FKBP genes were identified from the genome of s.litura and were clustered into 2 distinct groups,according to their subcellular localization,with FKBP13 and FKBP14 belonging to the endoplasmic reticulum(ER)group and with the other members belonging to the cytoplasmic(Cy)group.All the CyFKBPs were significantly upregulated in the prepupal or pupal stages,with the opposite being observed for the ER group members.FK506 completely blocked the transfer of EcR to the nucleus under 20E induction,and significantly downregulated the transcriptional expression of many 20E signaling genes.A similar phenomenon was observed after RNA interference of2 CyFKBPs(FKBP45 and FKBP12b),but not for FKBP13.Taken together,our data indicate that the cytoplasmic FKBPs,especially FKBP45 and FKBP12b,mediate the nuclear localization of EcR,thereby regulating the 20E signaling and ultimately affecting molting and metamorphosis in insects.展开更多
Adolescent depression is increasingly recognized as a serious mental health disorder with distinct clinical and molecular features.Using single-nucleus RNA sequencing,we identified cell-specific transcriptomic changes...Adolescent depression is increasingly recognized as a serious mental health disorder with distinct clinical and molecular features.Using single-nucleus RNA sequencing,we identified cell-specific transcriptomic changes in the nucleus accumbens(NAc),particularly in astrocytes,of adolescent macaques exhibiting depressive-like behaviors.The level of diacylglycerol kinase beta was significantly reduced in neurons and glial cells of depressed macaques,while FKBP5 levels increased in glial cells.Disruption of GABAergic synapses and disruption of D-glutamine and D-glutamate metabolism were linked to depressive phenotypes in medium spiny neurons(MSNs)and subtypes of astrocytes.Communication pathways between astrocytes and D1/D2-MSNs were also disrupted,involving factors like bone morphogenetic protein-6 and Erb-B2 receptor tyrosine kinase-4.Bulk transcriptomic and proteomic analyses corroborated these findings,and FKBP5 upregulation was confirmed by qRT-PCR,western blotting,and immunofluorescence in the NAc of rats and macaques with chronic unpredictable mild stress.Our results highlight the specific roles of different cell types in adolescent depression in the NAc,offering potential targets for new antidepressant therapies.展开更多
Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of th...Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.展开更多
基金The authors thank the National 863 Program Foundation(2002AA233051)of China for support.
文摘Based on the structure of FK506, FKBP12 and calcineurin complex and the interactivecharacteristics of small molecular ligands with FKBPs, a series of L-1,4-thiazane-3-carboxylic acidderivatives as neuroimmunophilin ligands was designed and synthesized. The results of evaluationshow that compound N308 has a great promise as a candidate of neuroprotective andneuroregenerative agent.
基金supported by grants from the National Natural Science Foundation of China(grant no.32272523)the Natural Science Foundation of Guangdong Province,China(grant no.2023A1515010178)the Laboratory of Lingnan Modern Agriculture Project(grant no.NT2021003).
文摘Molting and metamorphosis are important physiological processes in insects that are tightly controlled by ecdysone receptor(EcR)through the 20-hydroxyecdysone(20E)signaling pathway.EcR is a steroid nuclear receptor(SR).Several FK506-binding proteins(FKBPs)have been identified from the mammal SR complex,and are thought to be involved in the subcellular trafficking of SR.However,their roles in insects are poorly understood.To explore whether FKBPs are involved in insect molting or metamorphosis,we injected an FKBP inhibitor(FK506)into a lepidopteran insect,Spodoptera litura,and found that molting was inhibited in 61.11%of the larvae,and that the time for larvae to pupate was significantly extended.A total of 10 FKBP genes were identified from the genome of s.litura and were clustered into 2 distinct groups,according to their subcellular localization,with FKBP13 and FKBP14 belonging to the endoplasmic reticulum(ER)group and with the other members belonging to the cytoplasmic(Cy)group.All the CyFKBPs were significantly upregulated in the prepupal or pupal stages,with the opposite being observed for the ER group members.FK506 completely blocked the transfer of EcR to the nucleus under 20E induction,and significantly downregulated the transcriptional expression of many 20E signaling genes.A similar phenomenon was observed after RNA interference of2 CyFKBPs(FKBP45 and FKBP12b),but not for FKBP13.Taken together,our data indicate that the cytoplasmic FKBPs,especially FKBP45 and FKBP12b,mediate the nuclear localization of EcR,thereby regulating the 20E signaling and ultimately affecting molting and metamorphosis in insects.
基金supported by STI2030-Major Projects(2022ZD0212900)the Joint Project of Chongqing Municipal Science and Technology Bureau and Chongqing Health Commission(2023CCXM003)+4 种基金the National Key Research and Development Program of China(2017YFA0505700)the National Natural Science Foundation of China(82271565 and 82301714)the China Postdoctoral Science Foundation(2023TQ0398,GZB20230916,2023MD734124)the Natural Science Foundation of Chongqing,China(CSTB2023NSCQ-BHX0106)the Postdoctoral Innovation Talents Support Program of Chongqing,China(2208013341918508).
文摘Adolescent depression is increasingly recognized as a serious mental health disorder with distinct clinical and molecular features.Using single-nucleus RNA sequencing,we identified cell-specific transcriptomic changes in the nucleus accumbens(NAc),particularly in astrocytes,of adolescent macaques exhibiting depressive-like behaviors.The level of diacylglycerol kinase beta was significantly reduced in neurons and glial cells of depressed macaques,while FKBP5 levels increased in glial cells.Disruption of GABAergic synapses and disruption of D-glutamine and D-glutamate metabolism were linked to depressive phenotypes in medium spiny neurons(MSNs)and subtypes of astrocytes.Communication pathways between astrocytes and D1/D2-MSNs were also disrupted,involving factors like bone morphogenetic protein-6 and Erb-B2 receptor tyrosine kinase-4.Bulk transcriptomic and proteomic analyses corroborated these findings,and FKBP5 upregulation was confirmed by qRT-PCR,western blotting,and immunofluorescence in the NAc of rats and macaques with chronic unpredictable mild stress.Our results highlight the specific roles of different cell types in adolescent depression in the NAc,offering potential targets for new antidepressant therapies.
基金supported by the National Natural Science Foundation of China(82404995,82404890)Research Funds of Center for Xin’an Medicine and Modernization of Traditional Chinese Medicine of IHM(2023CXMMTCM013)+3 种基金Scientific Research Program of Anhui Provincial Department of Education(2024AH051036,2024AH040137,2024AH051044)Excellent Funding for Academic and Scientific Research Activities for Academic and Technological Leaders in Anhui Province(2022D317)Key Research and Development Plan of Anhui Province(202104j07020004)Anhui University of Chinese Medicine Talent Support Program(DT2300000173).
文摘Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.
