RESUMEN
Due to the medicinal importance of the flowers of Xianglei type (XL) Lonicera macranthoides, it is important to understand the molecular mechanisms that underlie their development. In this study, we elucidated the transcriptomic and metabolomic mechanisms that underlie the flower development mechanism of two L. macranthoides varieties. In this study, 3435 common differentially expressed unigenes (DEGs) and 1138 metabolites were identified. These common DEGs were mainly enriched in plant hormone signal transduction pathways. Metabolomic analysis showed that amino acids were the main metabolites of differential accumulation in wild-type (WT) L. macranthoides, whereas in XL, they were flavonoids and phenylalanine metabolites. Genes and transcription factors (TFs), such as MYB340, histone deacetylase 1 (HDT1), small auxin-up RNA 32 (SAUR32), auxin response factor 6 (ARF6), PIN-LIKES 7 (PILS7), and WRKY6, likely drive metabolite accumulation. Plant hormone signals, especially auxin signals, and various TFs induce downstream flower organ recognition genes, resulting in a differentiation of the two L. macranthoides varieties in terms of their developmental trajectories. In addition, photoperiodic, autonomous, and plant hormone pathways jointly regulated the L. macranthoides corolla opening. SAUR32, Arabidopsis response regulator 9 (ARR9), Gibberellin receptor (GID1B), and Constans-like 10 (COL10) were closely related to the unfolding of the L. macranthoides corolla. These findings offer valuable understanding of the flower growth process of L. macranthoides and the excellent XL phenotypes at the molecular level. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-04019-1.
RESUMEN
This study aims to explore the molecular regulatory mechanism of the differential accumulation of flavonoids between 'Xianglei' and the wild type of Lonicera macranthoides. The flowers, stems, and leaves of the two varieties of L. macranthoides were collected. Ultra-performance liquid chromatography-mass spectrometry(UPLC-MS) and high-throughput sequencing(RNA-seq) were employed to screen out the differential flavonoids, key differentially expressed genes(DEGs) and transcription factors(TFs). Fourteen DEGs were randomly selected for verification by qRT-PCR. The results showed that a total of 17 differential flavonoids were obtained, including naringin chalcone, apigenin, and quercetin. The transcriptomic analysis predicted 19 DEGs associated with flavonoids, including 2 genes encoding chitin synthase(CHS) and 3 genes encoding chalcone isomerase(CHI). The regulatory network analysis and weighted gene co-expression network analysis(WGCNA) screen out the key enzyme genes CHS1, FLS1, and HCT regulating the accumulation of flavonoids. MYB12 and LBD4 may be involved in the biosynthesis of flavonoids by regulating the expression of key enzyme genes CHS1, FLS1, and HCT. The qRT-PCR and RNA-seq results were similar regarding the expression patterns of the 14 randomly selected DEGs. This study preliminarily analyzed the transcriptional regulatory mechanism for the differential accumulation of flavonoids in the two varieties of L. macranthoides and laid a foundation for further elucidating the regulatory effects of key enzyme genes and TFs on the accumulation of flavonoids.
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Flavonoides , Regulación de la Expresión Génica de las Plantas , Lonicera , Metabolómica , Transcriptoma , Lonicera/genética , Lonicera/metabolismo , Lonicera/química , Flavonoides/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Perfilación de la Expresión Génica , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
OBJECTIVE: To investigate the effect of isorhamnetin on the pathology of rheumatoid arthritis (RA). METHODS: Tumor necrosis factor (TNF)- α -induced fibroblast-like synoviocytes (FLS) was exposed to additional isorhamnetin (10, 20 and 40 µ mol/L). Overexpression vectors for matrix metalloproteinase-2 (MMP2) or MMP9 or SRC were transfected to explore their roles in isorhamnetin-mediated RA-FLS function. RA-FLS viability, migration, and invasion were evaluated. Moreover, a collagen-induced arthritis (CIA) rat model was established. Rats were randomly divided to sham, CIA, low-, medium-, and high-dosage groups using a random number table (n=5 in each group) and administed with normal saline or additional isorhamnetin [2, 10, and 20 mg/(kg·day)] for 4 weeks, respectively. Arthritis index was calculated and synovial tissue inflammation was determined in CIA rats. The levels of MMP2, MMP9, TNF-α, interleukin-6 (IL-6), and IL-1 ß, as well as the phosphorylation levels of SRC, extracellular regulated kinase (ERK), and cyclic adenosine monophosphate response element-binding (CREB), were detected in RA-FLS and synovial tissue. Molecular docking was also used to analyze the binding of isorhamnetin to SRC. RESULTS: In in vitro studies, isorhamnetin inhibited RA-FLS viability, migration and invasion (P<0.05). Isorhamnetin downregulated the levels of MMP2, MMP9, TNF-α, IL-6, and IL-1 ß in RA-FLS (P<0.05). The overexpression of either MMP2 or MMP9 reversed isorhamnetin-inhibited RA-FLS migration and invasion, as well as the levels of TNF-α, IL-6, and IL-1 ß (P<0.05). Furthermore, isorhamnetin bound to SRC and reduced the phosphorylation of SRC, ERK, and CREB (P<0.05). SRC overexpression reversed the inhibitory effect of isorhamnetin on RA-FLS viability, migration and invasion, as well as the negative regulation of MMP2 and MMP9 (P<0.05). In in vivo studies, isorhamnetin decreased arthritis index scores (P<0.05) and alleviated synovial inflammation. Isorhamnetin reduced the levels of MMP2, MMP9, TNF-α, IL-6, and IL-1 ß, as well as the phosphorylation of SRC, ERK, and CREB in synovial tissue (P<0.05). Notably, the inhibitory effect of isorhamnetin was more pronounced at higher concentrations (P<0.05). CONCLUSION: Isorhamnetin exhibited anti-RA effects through modulating SRC/ERK/CREB and MMP2/MMP9 signaling pathways, suggesting that isorhamnetin may be a potential therapeutic agent for RA.
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Artritis Experimental , Artritis Reumatoide , Quercetina/análogos & derivados , Ratas , Animales , Metaloproteinasa 2 de la Matriz/metabolismo , Familia-src Quinasas/metabolismo , Familia-src Quinasas/farmacología , Familia-src Quinasas/uso terapéutico , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Simulación del Acoplamiento Molecular , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Membrana Sinovial/metabolismo , Membrana Sinovial/patología , Inflamación/patología , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/patología , Células Cultivadas , Fibroblastos , Proliferación CelularRESUMEN
Rheumatoid arthritis(RA), as a chronic autoimmune disease, has a high incidence and disability rate, causing significant suffering to patients. Due to its complex pathogenesis, it has not been fully elucidated to date, and its treatment remains a challenging problem in the medical field. Although western medicine treatment options have certain efficacy, they require prolonged use and are expensive. Additionally, they carry risks of multiple infections and adverse reactions like malignancies. The Chinese herbal medicine Rhododendron molle is commonly used in folk medicine for its properties of dispelling wind, removing dampness, calming nerves, and alleviating pain in the treatment of diseases like rheumatic bone diseases. In recent years, modern clinical and pharmacological studies have shown that the diterpenoids in R. molle are effective components, exhibiting immune-regulatory, anti-inflammatory, and analgesic effects. This makes it a promising candidate for treating RA with a broad range of potential applications. However, R. molle has certain toxic properties that hinder its clinical application and lead to the wastage of its resources. This study reviewed recent research progress on the mechanism of R. molle in preventing and treating RA, focusing on its chemical components, anti-inflammatory and analgesic properties and summarized the adverse reactions associated with R. molle, aiming to offer new ideas for finding natural remedies for RA and methods to reduce toxicity while enhancing the effectiveness of R. molle. The study seeks to clarify the safety and efficacy of R. molle and its extracts, providing a theoretical basis for its application prospects and further promoting the development and utilization of R. molle resources.
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Artritis Reumatoide , Diterpenos , Rhododendron , Humanos , Rhododendron/química , Artritis Reumatoide/tratamiento farmacológico , Antiinflamatorios , Diterpenos/farmacología , AnalgésicosRESUMEN
As a large family of transcription factors, the MYB family plays a vital role in regulating flower development. We studied the MYB family members in Lonicera macranthoides for the first time and identified three sequences of 1R-MYB, 47 sequences of R2R3-MYB, two sequences of 3R-MYB, and one sequence of 4R-MYB from the transcriptome data. Further, their physicochemical properties, conserved domains, phylogenetic relationship, protein structure, functional information, and expression were analyzed. The results show that the 53 MYB transcription factors had different conserved motifs, physicochemical properties, structures, and functions in wild type and 'Xianglei' cultivar of L. macranthoides, indicating their conservation and diversity in evolution. The transcript level of LmMYB was significantly different between the wild type and 'Xianglei' cultivar as well as between flowers and leaves, and some genes were specifically expressed. Forty-three out of 53 LmMYB sequences were expressed in both flowers and leaves, and 9 of the LmMYB members showed significantly different transcript levels between the wild type and 'Xianglei' cultivar, which were up-regulated in the wild type. The results provide a theoretical basis for further studying the specific functional mechanism of the MYB family.
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Lonicera , Factores de Transcripción , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Lonicera/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
The present study explored the consistency of the content proportions of active components of Aurantii Fructus and analyzed the influencing factors based on three-dimensional multi-component analysis. A total of 839 Aurantii Fructus samples in 65 research articles were analyzed using the three-dimensional multi-component analysis mode. The content data of flavonoid components(naringin, hesperidin, neohesperidin, narirutin, and nobiletin), coumarin components(meranzin and gluconolactone), and alkaloid(synephrine) in 386 samples which met the criteria of 2020 edition of the Chinese Pharmacopoeia were extracted and adjusted to percentages, and the content ratios between components were calculated. The influencing factors of Aurantii Fructus quality were analyzed. The results showed content ratios of components as follows: neohesperidinâ¶naringin in the range of 0.4-1.2; narirutinâ¶naringin in the range of 0.02-0.16; hesperidinâ¶naringin in the range of 0.01-0.3; nobiletinâ¶naringin in the range of 0.000 588 3-0.069 68; synephrineâ¶naringin in the range of 0.02-0.042; gluconolactoneâ¶naringin in the range of 0.001-0.01; meranzinâ¶naringin in the range of 0.000 4-0.035. The quality of Aurantii Fructus was closely related to the origin, variety, harvesting time, and processing method of medicinal materials. Harvesting time had a greater impact on the quality of Aurantii Fructus, and the origin and variety had a certain impact on the quality of Aurantii Fructus. The findings of this study indicated that the ratios between flavonoid components, flavonoids and coumarin components, and flavonoids and alkaloids fluctuated. The production base should optimize the varieties, harvesting period, and processing methods of Aurantii Fructus to provide a scientific basis for the production of high-quality Aurantii Fructus.
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Citrus , Medicamentos Herbarios Chinos , Flavonoides/análisis , Frutas/química , Cumarinas/análisis , Cromatografía Líquida de Alta Presión/métodosRESUMEN
BACKGROUND: Lonicera macranthoides Hand.-Mazz. is an important medicinal plant. Xianglei-type (XL) L. macranthoides was formed after many years of cultivation by researchers on the basis of the natural mutant. The corolla of L. macranthoides XL remains unexpanded and its flowering period is nearly three times longer than that of wild-type (WT) plants. However, the molecular mechanism behind this desirable trait remains a mystery. OBJECTIVE: To understand the floral phenotype differences between L. macranthoides and L. macranthoides XL at the molecular level. METHODS: Transcriptome analysis was performed on L. macranthoides XL and WT. One DEG was cloned by RT-PCR amplification and selected for qRT-PCR analysis. RESULTS: Transcriptome analysis showed that there were 5603 differentially expressed genes (DEGs) in XL vs. WT. Enrichment analysis of DEGs showed that pathways related to plant hormone signal transduction were significantly enriched. We identified 23 key genes in ethylene biosynthesis and signal transduction pathways. The most abundant were the ethylene biosynthesis DEGs. In addition, the open reading frames (ORFs) of WT and XL ETR2 were successfully cloned and named LM-ETR2 (GenBank: MW334978) and LM-XL-ETR2 (GenBank: MW334978), respectively. qRT-PCR at different flowering stages suggesting that ETR2 acts in the whole stage of flower development of WT and XL. CONCLUSIONS: This study provides new insight into the molecular mechanism that regulates the development of special traits in the flowers of L. macranthoides XL. The plant hormone ethylene plays an important role in flower development and flowering duration prolongation in L. macranthoides. The ethylene synthesis gene could be more responsible for the flower phenotype of XL. The genes identified here can be used for breeding and improvement of other flowering plants after functional verification.
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Lonicera , Lonicera/genética , Lonicera/metabolismo , Reguladores del Crecimiento de las Plantas/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Fitomejoramiento , Perfilación de la Expresión Génica , Etilenos/metabolismoRESUMEN
BACKGROUND: Rheumatoid arthritis (RA) is a chronic inflammatory disease with a high rate of disability accompanied by various complications. The pathogenesis of RA is complex with multiple targets and links. This study aims to investigate pharmacological effects and mechanism of Rhodojaponin III in RA. METHODS: The bovine type II collagen-induced arthritis (CIA) rat model and tumor necrosis factor-alpha (TNF-α) induced human umbilical vein endothelial cells (HUVECs) model were constructed. Different concentrations of Rhodojaponin III were utilized for intervention. The progression of CIA was assessed by the arthritis index (AI). Pathological changes in knee joints and synovium were observed. The expressions of angiogenesis-related cytokines were detected. The proliferation, migration, invasion, and angiogenesis of HUVECs were detected. The levels of pro-inflammatory cytokines were determined. The expressions of nuclear factor kappa B-inducing kinase (NIK) pathway-related proteins were analyzed. The binding of Rhodojaponin III to NIK was simulated by molecular docking. RESULTS: Rhodojaponin III suppressed cartilage damage and bone erosion in the knee joints. Rhodojaponin III inhibited expressions of platelet endothelial cell adhesion molecule-1 (CD31) and vascular endothelial cell growth factor (VEGF) to decrease vascular density. Rhodojaponin III suppressed the proliferation, migration, invasion, and angiogenesis of HUVECs, and decreased the levels of interleukin (IL)-6, IL-1ß, and TNF-α. Molecular docking showed that Rhodojaponin III could spontaneously bind to NIK. Rhodojaponin III decreased the expression of NIK, p52, and C-X-C motif chemokine ligand 12 (CXCL12) and the phosphorylation level of IκB kinase-alpha (IKKα) in the synovium of CIA rats and TNF-α-induced HUVECs. NIK overexpression reversed the inhibitory effect of Rhodojaponin III on activation of the NIK/NF-κB pathway, migration, invasion, and angiogenesis of HUVECs, and the secretion of pro-inflammatory cytokines. CONCLUSIONS: Rhodojaponin III affected the angiogenesis and inflammation of CIA rats and TNF-α-induced HUVECs by regulating the NIK/IKKα/CXCL12 pathway. These findings suggest that Rhodojaponin III has potential as a therapeutic agent for RA. Further studies are needed to explore its precise mechanism of action and evaluate its clinical efficacy.
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Artritis Experimental , Artritis Reumatoide , Ratas , Humanos , Animales , Bovinos , Factor de Necrosis Tumoral alfa , Quinasa I-kappa B , Simulación del Acoplamiento Molecular , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Citocinas/metabolismo , FN-kappa B/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/metabolismoRESUMEN
In order to explore the functions of genes of key rate-limiting enzymes chalcone isomerase(CHI) and chalcone synthase(CHS) in the biosynthesis of flavonoids in Lonicera macranthoides, this study screened and cloned the cDNA sequences of CHI and CHS genes from the transcriptome data of conventional variety and 'Xianglei' of L. macranthoides. Online bioinformatics analysis software was used to analyze the characteristics of the encoded proteins, and quantitative reverse-transcription polymerase chain reaction(qRT-PCR) to detect the expression of CHI and CHS in different parts of the varieties at different flowering stages. The content of luteo-loside was determined by high performance liquid chromatography(HPLC) and the correlation with the expression of the two genes was analyzed. The results showed that the CHI and CHS of the two varieties contained a 627 bp and 1170 bp open reading frame(ORF), respectively, and the CHI protein and CHS protein were stable, hydrophilic, and non-secretory. qRT-PCR results demonstrated that CHI and CHS of the two varieties were differentially expressed in stems and leaves at different flowering stages, particularly the key stages. Based on HPLC data, luteoloside content was in negative correlation with the relative expression of the genes. Thus, CHI and CHS might regulate the accumulation of flavonoids in L. macranthoides, and the specific functions should be further studied. This study cloned CHI and CHS in L. macranthoides and analyzed their expression for the first time, which laid a basis for investigating the molecular mechanism of the differences in flavonoids such as luteoloside in L. macranthoides and variety breeding.
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Chalcona , Lonicera , Aciltransferasas/genética , Aciltransferasas/metabolismo , Clonación Molecular , Liasas Intramoleculares , Lonicera/genética , Lonicera/metabolismo , FitomejoramientoRESUMEN
To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 µm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 µL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 µL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½ß) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
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Ácidos Cumáricos/farmacocinética , Medicamentos Herbarios Chinos/farmacocinética , Glucósidos/farmacocinética , Iridoides/farmacocinética , Estilbenos/farmacocinética , Administración Oral , Animales , Disponibilidad Biológica , Ácidos Cumáricos/administración & dosificación , Ácidos Cumáricos/sangre , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/análisis , Glucósidos/administración & dosificación , Glucósidos/sangre , Iridoides/administración & dosificación , Iridoides/sangre , Masculino , Ratas , Ratas Sprague-Dawley , Estilbenos/administración & dosificación , Estilbenos/sangreRESUMEN
OBJECTIVE: To study on volatice oil from Atractylodes macrosephala Koidz with different distill methods and find the better method. METHODS: GC-MS was used to analyze the chemical constituents of volatice oil from Atractylodes macrocephala Koidz with different distill methods. RESULTS: The extraction rates of volatice oil with steam distillation was 1.01%, the components of the oil were examined by GC-MS, 15 of the 18 were identified. The extraction rates of volatice oil with ultrasonic wave was 1.60%, the components examined, 20 of the 24 were identified. The extraction rates of volatice oil with SFE-CO2 was 2.32%, the components examined, 37 of the 49 were identified. Atractylon was the highest one. There were 12 common components in the identified ones. CONCLUSION: The components of volatice oil from Atractylodes macrocephala Koidz with different distill methods have difference but similarities, it can provide a method for Atractylodes macrocephala Koidz's quality control. The extraction rates is higher and the components are more with the method of SFE-CO2.
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Atractylodes/química , Aceites Volátiles/aislamiento & purificación , Plantas Medicinales/química , Tecnología Farmacéutica/métodos , Dióxido de Carbono , Cromatografía con Fluido Supercrítico/métodos , Cromatografía de Gases y Espectrometría de Masas/métodos , Aceites Volátiles/química , Sesquiterpenos/análisis , Sesquiterpenos/aislamiento & purificación , Vapor , UltrasonidoRESUMEN
OBJECTIVE: To determine the best manure scheme of lily (Lilium lancifolium). METHODS: Determining lilypolysaccharide and phosphatide contents through phenol hydrate-sulfuric acid and molybdenum blue colorimetric method. RESULTS: The content of efficacious composition in applying fertilizer on the leaves is higher than average and possium fertilizer can increase lilypolysaccharide content. CONCLUSION: Possium fertilizer is important in early stage and leaf fertilizer can improve lily quality.
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Fertilizantes , Lilium , Estiércol , Plantas Medicinales , Fertilizantes/clasificación , Lilium/crecimiento & desarrollo , Nitrógeno , Fosfolípidos/análisis , Fosfolípidos/aislamiento & purificación , Plantas Medicinales/crecimiento & desarrollo , Polisacáridos/análisis , Polisacáridos/aislamiento & purificación , Potasio , Control de Calidad , Espectrofotometría Ultravioleta , Factores de Tiempo , UreaRESUMEN
OBJECTIVE: To exploit and utilize reasonably the abundant natural resources by appraising the medicinal purposes of Lonicera macranthoides Hands-Mazz. through the content of the useful chemical constitutents chlorogenic acid. METHOD: The content of chlorogenic acid in the flowers of L. macranthoides Hands-Mazz in Human and L. japonica Thunb. in Henan and Shandong was compared by HPLC. RESULT: The contents of chlorogenic acid were 4.00% and 4.52% in L. macrathoides from Longhui and Xinning County of Hunan respectively. The contents were 2.20% and 2.46% in L. japonica from Shangdong and Henan respectively. CONCLUSION: The content of chlorogenic acid in L. macranthoides was higher than that in Lonicera japonica.