ABSTRACT
To study the mechanism of anthocyanin-biosynthesis regulation, we examined light-regulated gene expression involved in anthocyanin biosynthesis in purple grains of wheat. Ten kinds of anthocyanins were identified from a purple-grained wheat cultivar by HPLC-ESI-MS/MS analysis. Libraries constructed from the total RNA of purple grains under light (L) or dark (D) conditions for 15 or 20 days were sequenced. In total, 1874 differentially expressed genes (DEGs) were identified in L20 vs L15, 1432 DEGs were identified in D20 vs D15, 862 DEGs were identified in D15 vs L15, and 1786 DEGs were identified in D20 vs L20. DEG functional enrichments suggested that light-signal transduction is critical to anthocyanin biosynthesis. The 911 DEGs referred to as light-regulated DEGs (LDEGs) involved a number of genes in anthocyanin biosynthesis, transcription regulation, sugar- and calcium-signaling pathways, and hormone metabolism. These findings laid the foundation for future studies on the regulatory mechanisms of anthocyanin biosynthesis in purple grains of wheat.
Subject(s)
Anthocyanins/biosynthesis , Plant Proteins/genetics , Triticum/genetics , Gene Expression Regulation, Plant/radiation effects , Light , Plant Proteins/metabolism , Seeds/genetics , Seeds/metabolism , Seeds/radiation effects , Transcriptome/radiation effects , Triticum/metabolism , Triticum/radiation effectsABSTRACT
Extracellular matrix (ECM) degrading enzymes, including matrix metalloproteinases (MMPs), are critical for cartilage destruction in the progression of osteoarthritis (OA). Thus, identifying novel drugs, which suppress the synthesis of MMPs may facilitate the treatment of OA. The cytotoxicity of lycorine was determined using a CCK8 assay. The effects of lycorine on IL1ßinduced upregulation of MMPs and activation of mitogenactivated protein kinase pathways were detected by western blot analysis and reverse transcriptionquantitative polymerase chain reaction. Hematoxylin and eosin staining and Safranin O staining were used to evaluate the effect of lycorine in a mouse anterior cruciate ligament transection model. In the present study, it was demonstrated for the first time, to the best of our knowledge, that lycorine (LY) suppressed interleukin1ß (IL1ß)induced synthesis of MMP3 and MMP13 in vitro. Molecular analysis revealed that LY abrogated the phosphorylation of cJun Nterminal kinase (JNK) and the activation of the nuclear factor (NF)κB signaling pathway caused by IL1ß stimulation. In addition, in vivo experiments in a mouse anterior cruciate ligament transection model confirmed the protective role of LY on cartilage. Taken together, the data obtained in the present study demonstrated that LY suppressed the IL1ßinduced expression of MMP3 and MMP13 through inhibition of the JNK and NFκB pathways, suggesting that LY may be used as a potential drug for the treatment of OA.
Subject(s)
Amaryllidaceae Alkaloids/therapeutic use , Anti-Inflammatory Agents/therapeutic use , Chondrocytes/drug effects , Interleukin-1beta/immunology , Matrix Metalloproteinase 13/immunology , Matrix Metalloproteinase 3/immunology , Osteoarthritis/drug therapy , Phenanthridines/therapeutic use , Protective Agents/therapeutic use , Amaryllidaceae/chemistry , Amaryllidaceae Alkaloids/chemistry , Amaryllidaceae Alkaloids/pharmacology , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Cartilage, Articular/drug effects , Cartilage, Articular/immunology , Cartilage, Articular/pathology , Cell Survival/drug effects , Cells, Cultured , Chondrocytes/immunology , Chondrocytes/pathology , Disease Models, Animal , Male , Mice , Mice, Inbred C57BL , NF-kappa B/immunology , Osteoarthritis/immunology , Osteoarthritis/pathology , Phenanthridines/chemistry , Phenanthridines/pharmacology , Protective Agents/chemistry , Protective Agents/pharmacology , Rats , Rats, Sprague-DawleyABSTRACT
Osteoclasts play an important role in diseases involving bone loss. In this study, we assessed the effect of a plant-derived natural alkaloid (lycorine, or LY) on osteoclastogenesis in vitro and in vivo. Our in vitro study showed that receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis could be inhibited by LY; this effect was due to inhibition of mitogen-activated protein kinase (MAPK) signalling via MAP kinase kinases (MKKs). The MAPK agonist anisomycin could partially rescue the inhibitory effect of LY. Furthermore, LY also played a protective role in both a murine ovariectomy (OVX)-induced osteoporosis model and a titanium particle-induced osteolysis model. These results confirmed that LY was effective in preventing osteoclast-related diseases in vivo. In conclusion, our results show that LY is effective in suppressing osteoclastogenesis and therefore could be used to treat OVX-induced osteoporosis and wear particle-induced osteolysis.
Subject(s)
Amaryllidaceae Alkaloids/pharmacology , Bone Density Conservation Agents/pharmacology , Osteogenesis/drug effects , Osteolysis/prevention & control , Osteoporosis/prevention & control , Phenanthridines/pharmacology , RANK Ligand/genetics , Animals , Bone Marrow/drug effects , Cell Differentiation/drug effects , Cell Survival/drug effects , Disease Models, Animal , Female , Gene Expression Regulation , MAP Kinase Kinase 4/genetics , MAP Kinase Kinase 4/metabolism , MAP Kinase Kinase 7/genetics , MAP Kinase Kinase 7/metabolism , Macrophages/drug effects , Macrophages/metabolism , Macrophages/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Osteoclasts/drug effects , Osteoclasts/metabolism , Osteoclasts/pathology , Osteolysis/chemically induced , Osteolysis/genetics , Osteolysis/pathology , Osteoporosis/genetics , Osteoporosis/metabolism , Osteoporosis/pathology , Ovariectomy , Primary Cell Culture , RANK Ligand/antagonists & inhibitors , RANK Ligand/metabolism , Signal Transduction , TitaniumABSTRACT
A new pyrrolidine derivative, 3-hydroxy-5-(hydroxymethyl)-4-(4'-hydroxyphenoxy)pyrrolidin-2-one (1), and eight known steroids, (22E,24R)-7beta,8beta-epoxy-3beta,5alpha,9alpha-trihydroxyergosta-22-en-6-one (2, a reassigned structure of (22E,24R)-5alpha,6alpha-epoxy-3beta,8beta,14alpha-trihydroxyergosta-22-en-7-one), (22E,24R)-3beta,5alpha,9alpha-trihydroxyergosta-7,22-dien-6-one (3), (22E,24R)-3beta,5alpha-dihydroxyergosta-7,22-dien-6-one (4), (22E,24R)-ergosta-7,22-dien-3beta/,5alpha,6beta-triol (5), (22E,24R)-ergosta-5,22-dien-3beta-ol (6), (22E,24R)-5alpha,8alpha-epidioxyergosta-6,22-dien-3beta-ol (7), (22E,24R)-5alpha,8alpha-epidioxyergosta-6,9(11),22-trien-3beta-ol (8), and (22E,24R)-1(10 --> 6)-abeo-ergosta-5,7,9,22-tetraen-3alpha-ol (9), were isolated from the cultures of Gibberella zeae, an endophytic fungus isolated from the marine green alga Codium fragile. Their structures and relative stereochemistry were elucidated by 1D, 2D NMR and mass spectroscopic techniques. Compound 1 showed cytotoxicity against A-549 and BEL-7402 cell lines.
