To screen for interacting proteins of the rice thylakoid lumen protein OsTLP16.5(encoded by Os06g0705100),which will help to elucidate the molecular regulatory mechanisms of OsTLP16.5 in rice.Total RNA was extracted f...To screen for interacting proteins of the rice thylakoid lumen protein OsTLP16.5(encoded by Os06g0705100),which will help to elucidate the molecular regulatory mechanisms of OsTLP16.5 in rice.Total RNA was extracted from rice to construct a cDNA library.A yeast two-hybrid(Y2H)bait vector,pGBKT7-OsTLP16.5,was constructed to screen candidate target proteins using the yeast two-hybrid system,followed by interaction validation.Large-scale screening was performed using next-generation sequencing(NGS)technology.The bait vector pGBKT7-OsTLP16.5 was successfully constructed and verified to be free of autoactivation.A library screen identified 32 potential interacting proteins,of which 3 showed positive interactions after validation.NGS technology revealed 977 potential interacting proteins,including 51 chloroplast-associated proteins.A rice cDNA library was successfully constructed,and interacting proteins of OsTLP16.5 were identified,providing a foundation for studying how OsTLP16.5 functions in rice.展开更多
Stem-leaf saponins from Panax notoginseng(SLSP)comprise numerous PPD-type saponins with diverse pharmacological properties;however,their role in Parkinson's disease(PD),characterized by microglia-mediated neuroinf...Stem-leaf saponins from Panax notoginseng(SLSP)comprise numerous PPD-type saponins with diverse pharmacological properties;however,their role in Parkinson's disease(PD),characterized by microglia-mediated neuroinflammation,remains unclear.This study evaluated the effects of SLSP on suppressing microglia-driven neuroinflammation in experimental PD models,including the 1-methyl-4-phenylpyridinium(MPTP)-induced mouse model and lipopolysaccharide(LPS)-stimulated BV-2 microglia.Our findings revealed that SLSP mitigated behavioral impairments and excessive microglial activation in models of PD,including MPTP-treated mice.Additionally,SLSP inhibited the upregulation of inducible nitric oxide synthase(i NOS)and cyclooxygenase-2(COX2)and attenuated the phosphorylation of PI3K,protein kinase B(AKT),nuclear factor-κB(NFκB),and inhibitor of NFκB proteinα(IκBα)both in vivo and in vitro.Moreover,SLSP suppressed the production of inflammatory markers such as interleukin(IL)-1β,IL-6,and tumor necrosis factor alpha(TNF-α)in LPS-stimulated BV-2cells.Notably,the P2Y2R agonist partially reversed the inhibitory effects of SLSP in LPStreated BV-2 cells.These results suggest that SLSP inhibit microglia-mediated neuroinflammation in experimental PD models,likely through the P2Y2R/PI3K/AKT/NFκB signaling pathway.These novel findings indicate that SLSP may offer therapeutic potential for PD by attenuating microglia-mediated neuroinflammation.展开更多
文摘To screen for interacting proteins of the rice thylakoid lumen protein OsTLP16.5(encoded by Os06g0705100),which will help to elucidate the molecular regulatory mechanisms of OsTLP16.5 in rice.Total RNA was extracted from rice to construct a cDNA library.A yeast two-hybrid(Y2H)bait vector,pGBKT7-OsTLP16.5,was constructed to screen candidate target proteins using the yeast two-hybrid system,followed by interaction validation.Large-scale screening was performed using next-generation sequencing(NGS)technology.The bait vector pGBKT7-OsTLP16.5 was successfully constructed and verified to be free of autoactivation.A library screen identified 32 potential interacting proteins,of which 3 showed positive interactions after validation.NGS technology revealed 977 potential interacting proteins,including 51 chloroplast-associated proteins.A rice cDNA library was successfully constructed,and interacting proteins of OsTLP16.5 were identified,providing a foundation for studying how OsTLP16.5 functions in rice.
基金supported by the Educational Commission of Shanghai in China(No.2021LK114)the Organizational Key Research and Development Program of Shanghai University of Traditional Chinese Medicine(No.2023YZZ02)the Xinglin Young Talent Program at Shanghai University of Traditional Chinese Medicine(No.A1-U17205010430)。
文摘Stem-leaf saponins from Panax notoginseng(SLSP)comprise numerous PPD-type saponins with diverse pharmacological properties;however,their role in Parkinson's disease(PD),characterized by microglia-mediated neuroinflammation,remains unclear.This study evaluated the effects of SLSP on suppressing microglia-driven neuroinflammation in experimental PD models,including the 1-methyl-4-phenylpyridinium(MPTP)-induced mouse model and lipopolysaccharide(LPS)-stimulated BV-2 microglia.Our findings revealed that SLSP mitigated behavioral impairments and excessive microglial activation in models of PD,including MPTP-treated mice.Additionally,SLSP inhibited the upregulation of inducible nitric oxide synthase(i NOS)and cyclooxygenase-2(COX2)and attenuated the phosphorylation of PI3K,protein kinase B(AKT),nuclear factor-κB(NFκB),and inhibitor of NFκB proteinα(IκBα)both in vivo and in vitro.Moreover,SLSP suppressed the production of inflammatory markers such as interleukin(IL)-1β,IL-6,and tumor necrosis factor alpha(TNF-α)in LPS-stimulated BV-2cells.Notably,the P2Y2R agonist partially reversed the inhibitory effects of SLSP in LPStreated BV-2 cells.These results suggest that SLSP inhibit microglia-mediated neuroinflammation in experimental PD models,likely through the P2Y2R/PI3K/AKT/NFκB signaling pathway.These novel findings indicate that SLSP may offer therapeutic potential for PD by attenuating microglia-mediated neuroinflammation.
