BACKGROUND Sepsis-induced intestinal injury disrupts barrier function and exacerbates systemic inflammation,contributing to high mortality.AIM To investigate the mechanism by which sphinganine protects against sepsisi...BACKGROUND Sepsis-induced intestinal injury disrupts barrier function and exacerbates systemic inflammation,contributing to high mortality.AIM To investigate the mechanism by which sphinganine protects against sepsisinduced intestinal injury,focusing on macrophage polarization and toll like receptor 2(TLR2)/nuclear factor kappa-B(NF-κB)signaling.METHODS A cecal ligation and puncture sepsis model was established in mice(n=20/group).Treatments included sphinganine(15 mg/kg)and a TLR2 agonist[fibroblast-stimulating lipopeptide-1(FSL-1),10μg/kg].Serum markers[diamine oxidase(DAO),interleukin(IL)-1β,tumor necrosis factor(TNF)-α,IL-6]were measured by enzyme-linked immunosorbent assay.Intestinal injury and macrophage polarization[cluster of differentiation(CD)86^(+)M1,CD206^(+)M2]were assessed via hematoxylin and eosin and immunofluorescence staining.Proteomic analysis,molecular docking,and Western blot were used to evaluate TLR2/NF-κB signaling.Data were analyzed by analysis of variance and t-test.RESULTS Sphinganine significantly reduced serum levels of DAO(P<0.05),IL-1β,TNF-α,and IL-6(P<0.05),preserved intestinal crypt structure,and enhanced tight junction protein zonula occludens-1 expression.It promoted a shift from M1(CD86^(+))to M2(CD206^(+))macrophages.Proteomics identified TLR2 as the most differentially expressed protein,and molecular docking confirmed strong binding between sphinganine and TLR2(-4.3 kcal/mol).Sphinganine downregulated TLR2 and phosphorylated-NF-κB p65 expression(P<0.05),effects reversed by FSL-1.Total NF-κB p65 levels remained unchanged.CONCLUSION Sphinganine protects against sepsis-induced intestinal injury by inhibiting TLR2/NF-κB signaling,modulating macrophage polarization toward the M2 phenotype,and preserving intestinal barrier integrity.展开更多
Traditionally, herbal medicine is consumed by drinking decoctions produced by boiling herbs with water. The functional components of the decoction are heat stable. Small RNAs(sRNAs) were reported as a new class of fun...Traditionally, herbal medicine is consumed by drinking decoctions produced by boiling herbs with water. The functional components of the decoction are heat stable. Small RNAs(sRNAs) were reported as a new class of functional components in decoctions. However, the mechanisms by which sRNAs survive heat treatment of the decoction and enter cells are unclear.Previous studies showed that plant-derived exosome-like nanoparticles(ELNs), which we call botanosomes, could deliver therapeutic reagents in vivo. Here, we report that heat-stable decoctosomes(ELNs) from decoctions have more therapeutic effects than the decoctions in vitro and demonstrate therapeutic efficacy in vivo. Furthermore, sRNAs, such as HJT-sRNA-m7 and PGY-sRNA-6, in the decoctosome exhibit potent anti-fibrosis and anti-inflammatory effects, respectively. Decoctosome is comprised of lipids, chemical compounds, proteins, and s RNAs. A medical decoctosome mimic is called bencaosome. A single lipid sphinganine(d22:0) identified in the decoctosome was mixed and heated with the synthesized sRNAs to form the simplest bencaosome. This simple bencaosome structure was identified by critical micelle concentration(cmc) assay that sRNAs coassembled with sphinganine(d22:0) to form the lipid layers of vesicles. The heating process facilitates co-assembly of sRNAs and sphinganine(d22:0) until a steady state is reached. The artificially produced sphinganine-HJT-sRNA-m7 and sphinganinePGY-sRNA-6 bencaosomes could ameliorate bleomycin-induced lung fibrosis and poly(I:C)-induced lung inflammation, respectively, following oral administration in mice. Our study not only demonstrates that the herbal decoctosome may represent a combinatory remedy in precision medicine but also provides an effective oral delivery route for nucleic acid therapy.展开更多
Sphinganine or dihydrosphingosine （d18：0, DHS）, one of the most abundant free sphingoid Long Chain Base （LCB） in plants, has been recently shown to induce both cytosolic and nuclear calcium transient increases an...Sphinganine or dihydrosphingosine （d18：0, DHS）, one of the most abundant free sphingoid Long Chain Base （LCB） in plants, has been recently shown to induce both cytosolic and nuclear calcium transient increases and a correlated Programmed Cell Death （PCD） in tobacco BY-2 cells. In this study, in order to get deeper insight into the LCB signaling pathway leading to cell death, the putative role of Reactive Oxygen Species （ROS） has been investigated. We show that DHS triggers a rapid dose-dependent production of H2O2 that is blocked by diphenyleniodonium （DPI）, indicating the involvement of NADPH oxidase（s） in the process. In addition, while DPI does not block DHS-induced calcium increases, the ROS production is inhibited by the broad spectrum calcium channel blocker lanthanum （La^3＋）. Therefore, ROS production occurs downstream of DHS-induced Ca^2＋ transients. Interestingly, DHS activates expression of defense-related genes that is inhibited by both La^3＋ and DPI. Since DPI does not prevent DHS-induced cell death, these results strongly indicate that DHS-induced H2O2 production is not implicated in PCD mechanisms but rather would be associated to basal cell defense mechanisms.展开更多
基金Supported by the Natural Science of Minhang District of Shanghai,No.2023MH2085.
文摘BACKGROUND Sepsis-induced intestinal injury disrupts barrier function and exacerbates systemic inflammation,contributing to high mortality.AIM To investigate the mechanism by which sphinganine protects against sepsisinduced intestinal injury,focusing on macrophage polarization and toll like receptor 2(TLR2)/nuclear factor kappa-B(NF-κB)signaling.METHODS A cecal ligation and puncture sepsis model was established in mice(n=20/group).Treatments included sphinganine(15 mg/kg)and a TLR2 agonist[fibroblast-stimulating lipopeptide-1(FSL-1),10μg/kg].Serum markers[diamine oxidase(DAO),interleukin(IL)-1β,tumor necrosis factor(TNF)-α,IL-6]were measured by enzyme-linked immunosorbent assay.Intestinal injury and macrophage polarization[cluster of differentiation(CD)86^(+)M1,CD206^(+)M2]were assessed via hematoxylin and eosin and immunofluorescence staining.Proteomic analysis,molecular docking,and Western blot were used to evaluate TLR2/NF-κB signaling.Data were analyzed by analysis of variance and t-test.RESULTS Sphinganine significantly reduced serum levels of DAO(P<0.05),IL-1β,TNF-α,and IL-6(P<0.05),preserved intestinal crypt structure,and enhanced tight junction protein zonula occludens-1 expression.It promoted a shift from M1(CD86^(+))to M2(CD206^(+))macrophages.Proteomics identified TLR2 as the most differentially expressed protein,and molecular docking confirmed strong binding between sphinganine and TLR2(-4.3 kcal/mol).Sphinganine downregulated TLR2 and phosphorylated-NF-κB p65 expression(P<0.05),effects reversed by FSL-1.Total NF-κB p65 levels remained unchanged.CONCLUSION Sphinganine protects against sepsis-induced intestinal injury by inhibiting TLR2/NF-κB signaling,modulating macrophage polarization toward the M2 phenotype,and preserving intestinal barrier integrity.
基金supported by the National Natural Science Foundation of China (81788101)the Ministry of Science and Technology of China (2015CB553406)+1 种基金the National Natural Science Foundation of China (81490531)the CAMS Innovation Fund for Medical Sciences (2017-I2M-1-009)
文摘Traditionally, herbal medicine is consumed by drinking decoctions produced by boiling herbs with water. The functional components of the decoction are heat stable. Small RNAs(sRNAs) were reported as a new class of functional components in decoctions. However, the mechanisms by which sRNAs survive heat treatment of the decoction and enter cells are unclear.Previous studies showed that plant-derived exosome-like nanoparticles(ELNs), which we call botanosomes, could deliver therapeutic reagents in vivo. Here, we report that heat-stable decoctosomes(ELNs) from decoctions have more therapeutic effects than the decoctions in vitro and demonstrate therapeutic efficacy in vivo. Furthermore, sRNAs, such as HJT-sRNA-m7 and PGY-sRNA-6, in the decoctosome exhibit potent anti-fibrosis and anti-inflammatory effects, respectively. Decoctosome is comprised of lipids, chemical compounds, proteins, and s RNAs. A medical decoctosome mimic is called bencaosome. A single lipid sphinganine(d22:0) identified in the decoctosome was mixed and heated with the synthesized sRNAs to form the simplest bencaosome. This simple bencaosome structure was identified by critical micelle concentration(cmc) assay that sRNAs coassembled with sphinganine(d22:0) to form the lipid layers of vesicles. The heating process facilitates co-assembly of sRNAs and sphinganine(d22:0) until a steady state is reached. The artificially produced sphinganine-HJT-sRNA-m7 and sphinganinePGY-sRNA-6 bencaosomes could ameliorate bleomycin-induced lung fibrosis and poly(I:C)-induced lung inflammation, respectively, following oral administration in mice. Our study not only demonstrates that the herbal decoctosome may represent a combinatory remedy in precision medicine but also provides an effective oral delivery route for nucleic acid therapy.
文摘Sphinganine or dihydrosphingosine （d18：0, DHS）, one of the most abundant free sphingoid Long Chain Base （LCB） in plants, has been recently shown to induce both cytosolic and nuclear calcium transient increases and a correlated Programmed Cell Death （PCD） in tobacco BY-2 cells. In this study, in order to get deeper insight into the LCB signaling pathway leading to cell death, the putative role of Reactive Oxygen Species （ROS） has been investigated. We show that DHS triggers a rapid dose-dependent production of H2O2 that is blocked by diphenyleniodonium （DPI）, indicating the involvement of NADPH oxidase（s） in the process. In addition, while DPI does not block DHS-induced calcium increases, the ROS production is inhibited by the broad spectrum calcium channel blocker lanthanum （La^3＋）. Therefore, ROS production occurs downstream of DHS-induced Ca^2＋ transients. Interestingly, DHS activates expression of defense-related genes that is inhibited by both La^3＋ and DPI. Since DPI does not prevent DHS-induced cell death, these results strongly indicate that DHS-induced H2O2 production is not implicated in PCD mechanisms but rather would be associated to basal cell defense mechanisms.
