RESUMEN
Camellia oleifera, a major woody oil crop in China, produces tea oil rich in unsaturated fatty acids, earning it names like liquid gold and eastern olive oil. This study provides an integrated investigation of the transcriptome and lipidome within seeds at the maturing process across three C. oleifera varieties, revealing a significant relationship between fatty acid production and genes involved in lipid synthesis. Through transcriptomic analysis, 26,344 genes with varied expression were found. Functional enrichment analysis highlighted that pathways related to starch and sucrose metabolism, plant hormone signal transduction, and lipid accumulation were highly enriched among the differentially expressed genes. Coordinated high expression of key genes (ACCase, KAS I, KAS II, KAS III, KAR, HAD, EAR, SAD, LPAAT, LACS, DGAT, PDAT) during the late maturation stage contributes largely to high oil content. Additionally, expression variations of SAD and FADs among different varieties were explored. The analysis suggests that high expression of genes such as FAD3, FAD7, and FAD8 notably increased linolenic acid content. This research provides new insights into the molecular mechanisms of oil biosynthesis in C. oleifera, offering valuable references for improving yield and quality.
Asunto(s)
Camellia , Ácidos Grasos , Regulación de la Expresión Génica de las Plantas , Lipidómica , Proteínas de Plantas , Semillas , Transcriptoma , Camellia/metabolismo , Camellia/genética , Camellia/crecimiento & desarrollo , Camellia/química , Semillas/metabolismo , Semillas/crecimiento & desarrollo , Semillas/genética , Semillas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ácidos Grasos/metabolismo , Aceites de Plantas/metabolismo , Aceites de Plantas/química , ChinaRESUMEN
Hyperlipidemia refers to the abnormal levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL-C) and high-density lipoprotein (HDL-C) in peripheral blood circulation. It is a predominant risk factor underlying cardiovascular and cerebrovascular diseases, including coronary heart disease and atherosclerosis. Furthermore, it is also one of the most prevalent chronic diseases globally. Liujunzi Decoction is the basic prescription for the treatment of spleen and stomach diseases. It can tonify the spleen and qi, remove dampness, and reduce turbidity. Moreover, it is also clinically used for the treatment of spleen deficiency hyperlipidemia. However, its metabolites and therapeutic effect on spleen deficiency hyperlipidemia have not been comprehensively determined in vitro and in vivo. This study established a rat model of spleen deficiency hyperlipidemia by inducing starvation and satiety disorders, exhaustion swimming, and intragastric administration of the fat emulsion. To identify related metabolite changes and serum lipid composition, UPLC-Q-TOF-MS, PCA, and OPLS-DA lipidological methods were performed. The results demonstrated significant changes in rat's signs during the modeling process, which were consistent with the criteria for the syndrome differentiation of spleen deficiency in traditional Chinese medicine. Furthermore, this study identified 100 potential biomarkers in rat serum, of which 52 were associated with lipid synthesis, such as LPC, PC, PI, PE, PA, Cer, SM, etc. The pathways involved were glycerol phospholipid, sphingomyelin, and glycerol ester metabolisms. After the Liujunzi decoction intervention, 56 potential biomarkers were observed in the high-dose group, alleviating the metabolic spectrum imbalance by reducing metabolite levels. In addition, metabolic pathway disturbances were markedly improved. This study provides references for future studies on Liujunzi decoction and furnishes essential data for assessing the relationships between chemical constituents and pharmacological activities of Liujunzi decoction.
RESUMEN
BACKGROUND AND OBJECTIVE: Glycyrrhiza glabra L. (GG) and Strychnos nux-vomica L. (NV) are traditional Chinese medicines (TCMs). Changes in the chemical composition may occur before and after the GG-NV compatibility. Ultra-performance liquid chromatography Q-exactive Orbitrap mass spectrometry (UPLC-QE-Orbitrap-MS) was applied here to study the difference in the components of the GG and NV decoctions before and after they were combined. The changes in the chemical composition of GG and NV before and after the combination were determined. METHODS: The precise molecular weight, retention time, and fragment ion peak of the different components of the decoctions before and after compatibility were obtained through UPLC-QE-Orbitrap-MS. Differential analysis methods, such as principal component analysis, were used for comparison. RESULTS: In the positive ion mode, 200 new components were added, whereas six components were lost. In the negative ion mode, 144 new compounds were identified, whereas three components were missing. CONCLUSIONS: The compatibility difference between GG and NV was studied through UPLC-QE-Orbitrap-MS. The chemical composition of GG and NV changed before and after compatibility, and a class of compounds different from GG and NV was identified in the co-decoction. This study provides an experimental basis for subsequent research into detoxification mechanisms of the GG-NV combination and offers a new analytical method for investigating the compatibility of various other TCM pairs.
Asunto(s)
Medicamentos Herbarios Chinos , Glycyrrhiza , Espectrometría de Masas , Estricnina , Glycyrrhiza/química , Cromatografía Líquida de Alta Presión/métodos , Estricnina/análisis , Estricnina/química , Espectrometría de Masas/métodos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/análisis , Análisis de Componente PrincipalRESUMEN
This study comprehensively investigates the physiological and molecular regulatory mechanisms of Camellia oleifera seedlings under drought stress with a soil moisture content of about 30%, where exogenous abscisic acid (ABA) was applied via foliar spraying at concentrations of 50 µg/L, 100 µg/L, and 200 µg/L. The results demonstrated that appropriate concentrations of ABA treatment can regulate the physiological state of the seedlings through multiple pathways, including photosynthesis, oxidative stress response, and osmotic balance, thereby aiding in the restructuring of their drought response strategy. ABA treatment effectively activated the antioxidant system by reducing stomatal conductance and moderately inhibiting the photosynthetic rate, thus alleviating oxidative damage caused by drought stress. Additionally, ABA treatment promoted the synthesis of osmotic regulators such as proline, maintaining cellular turgor stability and enhancing the plant's drought adaptability. The real-time quantitative PCR results of related genes indicated that ABA treatment enhanced the plant's response to the ABA signaling pathway and improved disease resistance by regulating the expression of related genes, while also enhancing membrane lipid stability. A comprehensive evaluation using a membership function approach suggested that 50 µg/L ABA treatment may be the most-effective in mitigating drought effects in practical applications, followed by 100 µg/L ABA. The application of 50 µg/L ABA for 7 h induced significant changes in various biochemical parameters, compared to a foliar water spray. Notably, superoxide dismutase activity increased by 17.94%, peroxidase activity by 30.27%, glutathione content by 12.41%, and proline levels by 25.76%. The content of soluble sugars and soluble proteins rose by 14.79% and 87.95%, respectively. Additionally, there was a significant decrease of 31.15% in the malondialdehyde levels.
RESUMEN
The decoction turns into a complex multiphase system following exposure to high temperature and a complex chemical environment. However, the differences in the concentration of key active ingredients in different phase states and the release of drugs in sedimentary phase have yet to be elucidated. A simple ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous quantitative determination of brucine, strychnine, liquiritin, isoliquiritin, isoliquiritigenin and glycyrrhizic acid concentrations and it was applied to compare the content of different phases and measure the release characteristics of the sedimentary phase in "Glycyrrhiza glabra-Nux vomica" decoction (NGD). The results show that the method's selectivity, precision (intraday and interday ≤ 2%), matrix effect (101-108%), recovery and stability results were acceptable according to the guidelines. The method is sensitive and reliable. The content determination results show that the most toxic strychnine in the sedimentary phase accounted for 75.70% of the total components. The different components exhibited differential release in different media, and its components were released in the artificial intestinal fluid up to 81.02% in 12 h. Several components conformed to the primary kinetic model and the Ritger-Peppas model, and the most toxic compound exhibited slow release, thus conforming to the Ritger-Peppas model. This study provides a standard of reference for studies investigating reduction in toxicity of the combination of Glycyrrhiza glabra (Glycyrrhiza glabra L.) and Nux vomica (Strychnos nux-vomica L.).
Asunto(s)
Strychnos nux-vomica , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Ácido Glicirrínico/análisis , Semillas/química , Estricnina/química , Strychnos nux-vomica/química , Espectrometría de Masas en TándemRESUMEN
The present study established the spectrum-effect relationship model of flavonoids in Citri Reticulatae Pericarpium(CRP) from 15 batches of Liujunzi Decoction and statistically analyzed the correlation between chemical peaks and efficacy to identify the main effective components. HPLC fingerprints of flavonoids in CRP from 15 batches of Liujunzi Decoction were established. HPLC analysis was carried out on the Venusil XBP C_(18)(L) column(4.6 mm×250 mm, 5 µm) at 30 â with acetonitrile-water(containing 0.1% formic acid) as mobile phase for gradient elution, a flow rate of 1.0 mL·min~(-1), and detection wavelength of 300 nm to obtain chemical fingerprints. Additionally, the effects of flavonoids from CRP in 15 batches of Liujunzi Decoction on the content of GAS, MTL, and VIP, TFF3 mRNA expression, and percentage of CD3~+ T-cells of model rats with spleen deficiency were determined. The spectrum-effect relationship model was established by gray correlation analysis. The results showed that the main characteristic peaks with great contribution to the regulation of gastrointestinal tract were peak 16(vicenin-2), peak 63(sinensetin), peak 64(isosinensetin), peak 65(nobiletin), peak 67(3,5,6,7,8,3',4'-heptemthoxyflavone), peak 68(tangeretin), and peak 69(5-desmethylnobiletin). Therefore, there was a linear correlation between flavonoids from CRP in Liujunzi Decoction and the efficacy, and the medicinal effect was achieved by multi-component action. This study is expected to provide a new idea for exploring the material basis of the effect, i.e., regulating qi prior to replenishing qi, of CRP in Liujunzi Decoction.
Asunto(s)
Citrus , Bazo , Animales , Citrus/química , Medicamentos Herbarios Chinos , Flavonoides/química , Flavonoides/farmacología , Hormonas , ARN Mensajero/genética , RatasRESUMEN
Covalent organic frameworks (COFs) have attracted considerable attention in sample pretreatment because of their unique characteristics. However, the submicron or micron size of COFs has restricted their wider applications in solid-phase extraction (SPE). Herein, multiwalled nanotubes (MWNTs) were used as substrate materials to synthesize core-shell structured MWNTs@COFs composites (MWNTs@SNW-1) using a simple self-assembly method. The as-prepared MWNTs@SNW-1 composite exhibited a high BET surface area, good thermal stability, and good adsorption capacity. The MWNTs@SNW-1 composite was used as an adsorbent in cartridge-based SPE to extract four phytohormones before determining their levels by high-performance liquid chromatography. The experimental parameters affecting extraction efficiency, including the amount of adsorbents, solution pH, ionic strength, eluent type, and eluent volume, were investigated. The developed method showed a wide linear range (0.37-100 ng mL-1), low detection limits (0.11-0.32 ng mL-1), low limits of quantification (0.37-1.07 ng mL-1), high enrichment factors (45.9-49.3), and good reproducibility (<4.8%) for phytohormones. The developed analytical method was used to analyze trace phytohormones in fruit juices with good recoveries, highlighting the potential of the MWNTs@SNW-1 composite as an adsorbent in sample preparation.
Asunto(s)
Estructuras Metalorgánicas , Nanotubos , Adsorción , Cromatografía Líquida de Alta Presión , Jugos de Frutas y Vegetales , Límite de Detección , Reguladores del Crecimiento de las Plantas , Reproducibilidad de los Resultados , Extracción en Fase SólidaRESUMEN
To evaluate the detoxification effect of a combination of Radix Glycyrrhizae (GU) and Semen Strychni (SN) from toxicokinetics and drug tissue distribution perspectives, decoctions of processed SN and codecoction of SN and GU (SGN) were prepared, and an HPLC-ESI-MS/MS method was developed to monitor the severe exposure level in 1-month toxicokinetics and tissue distribution experiments to detect brucine and strychnine in rats. The toxicokinetic characteristics and tissue distribution before and after the addition of GU were analyzed. The method was successfully applied to evaluate the toxicokinetics and tissue distribution before and after the combination of SN and GU. The results show that GU decreased the blood concentration of toxic components in SN, and a double peak was observed in the drug time curve. The results of tissue distribution show that a combination of GU and SN significantly decreased the accumulation of toxic substances in metabolic organs and accelerated the clearance of toxic substances in the brain. These results provide a reference for the toxicity reduction mechanism of GU combined with SN.
Asunto(s)
Medicamentos Herbarios Chinos , Semillas , Espectrometría de Masas en Tándem , Administración Oral , Animales , Medicamentos Herbarios Chinos/toxicidad , Ratas , Semillas/toxicidad , Espectrometría de Masas en Tándem/métodos , Distribución Tisular , ToxicocinéticaRESUMEN
A novel polyethylene glycol-polycaprolactone-poly-l-tyrosine (MPEG-PCL-PTyr) amphiphilic triblock copolymer micelle was synthesized for the first time. 10-hydroxycamptothecin (HCPT) was embedded in MPEG-PCL-PTyr nanomicelles using the emulsion solvent evaporation method. A series of was conducted to confirm the structure of the compound and to evaluate the physical properties of the MPEG-PCL-PTyr nanomicelles. Cellular uptake, cytotoxicity, and apoptosis were assessed using flow cytometry and MTT assays. Confocal microscopy and flow cytometry results demonstrated that the nanocapsules carrying HCPT had significantly increased anti-tumor activity against HepG2 cells and could target HepG2 cell lysosomes with obvious liver targeting. In addition, the drug-loaded nanomicelles could significantly block the S phase of cancer cells and induce apoptosis; thus, they could be potential carriers for future 10-HCPT delivery and cancer treatment.
Asunto(s)
Micelas , Polímeros , Camptotecina/análogos & derivados , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos/métodos , Tamaño de la Partícula , Polietilenglicoles/química , Polímeros/químicaRESUMEN
AIM: Cantharidin (CTD), the major component of the anti-cancer medicine obtained from Mylabris cichorii, exerts good inhibitory effects on several cancers, such as liver and breast cancer. However, owing to its toxicity, its oral administration can cause various adverse effects, limiting its clinical applications. Therefore, the development of a novel nano-drug delivery system for CTD would be highly beneficial. METHODS: A nanostructured lipid carrier (NLC) was designed to actively target CTD to tumor cells using a hyaluronic acid (HA)-decorated copolymer (mPEG-NH2); the NLCs were called HA-mPEG-CTD-NLC. HA-mPEG was synthesized using amidation, and HA-mPEG-CTD-NLC was generated through ultrasonic emulsification in water. The mean hydrodynamic diameter of the particles was approximately 119.3â¯nm. RESULTS: Pharmacokinetic studies revealed that the half-life of HA-mPEG-CTD-NLC and its area under the curve were higher than those of a CTD solution. Further, the plasma clearance rate of HA-mPEG-CTD-NLC was 0.41 times that of the CTD solution, implying a significantly prolonged drug retention time in vivo. Fluorescence in vivo endo-microscopy and optical in vivo imaging revealed that HA-mPEG-CTD-NLC had superior cytotoxicity and targeting efficacy against SMMC-7721 cells. An evaluation of the in vivo anti-tumor activity showed that HA-mPEG-CTD-NLC significantly inhibited tumor growth and prolonged survival in tumor-bearing mice, with a tumor inhibition rate of 65.96%. CONCLUSIONS: Our results indicate that HA-mPEG-CTD-NLC may have great potential in liver cancer-targeted therapy.
Asunto(s)
Cantaridina/administración & dosificación , Ácido Hialurónico/química , Sistema de Administración de Fármacos con Nanopartículas , Polietilenglicoles/química , Animales , Cantaridina/farmacocinética , Línea Celular Tumoral , Femenino , Ácido Hialurónico/farmacocinética , Lípidos/química , Neoplasias Hepáticas/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos BALB C , Estructura Molecular , Sistema de Administración de Fármacos con Nanopartículas/farmacocinética , Polietilenglicoles/farmacocinética , Ratas Sprague-DawleyRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Citri Reticulatae Pericarpium (Rutaceae, CRP), commonly called as Chenpi () in Chinese, is most frequently used as a qi-regulating drug in thousands of Chinese medicine prescriptions. CRP is found mainly in major citrus-producing areas such as the Guangdong, Guangxi, Sichuan, Fujian, and Zhejiang Provinces of China. Since thousands of years in China, CRP has been used widely in clinical practice to treat nausea, vomiting, indigestion, anepithymia, diarrhea, cough, expectoration, and so on. Currently, CRP is listed in the Pharmacopoeia of the People's Republic of China. The present paper reviews the botany, ethnopharmacology, phytochemistry, pharmacology, quality control, and toxicology of CRP. MATERIALS AND METHODS: Information on CRP was gathered from various sources including the books on traditional Chinese herbal medicine; scientific databases including Elsevier, PubMed, and ScienceDirect; Baidu Scholar; CNKI; and others and from different professional websites. RESULTS: Approximately 140 chemical compounds have been isolated and identified from CRP. Among them, volatile oils and flavonoids are generally considered as the main bioactive and characteristic ingredients. CRP possesses wide pharmacological effects such as having a beneficial effect on the cardiovascular, digestive, and respiratory systems, antitumor, antioxidant, and anti-inflammatory properties; and a protective effect on the liver and nerve. Moreover, hesperidin is chosen as an indicator in the quantitative determination of CRP, and the quantity of aflatoxin in CRP must not exceed the standard limit mentioned in the pharmacopoeia. CONCLUSIONS: In brief, CRP has a warming nature, and hence, it can be used in harmony with a lot of medicines. CRP not only exhibits its effects individually but also aids other medicines exhibit a better effect. CRP can be consumed with tea, food, alcohol, and medicine. Irrespective of the form it is being consumed, CRP not only shows a synergistic effect but also has strengths on its own. Modern pharmacological studies have demonstrated that CRP has marked bioactivities, especially on the diseases of the digestive and respiratory systems. The bioactivities of CRP are useful for its clinical application and provide prospects for the development of drugs as well as food and health products for people. Although CRP is a commonly used drug in the traditional Chinese herbal prescription, there is an urgent need for further research on its synergistic effect with other herbs based on the compatibility theory of TCM, which would further increase our understanding on the compatibility theory of TCM.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Etnofarmacología , Medicina Tradicional China/métodos , Animales , Sinergismo Farmacológico , HumanosRESUMEN
OBJECTIVE: To construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration. METHODS: The lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods. RESULTS: The lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01). CONCLUSION: Lentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.
Asunto(s)
Disco Intervertebral/metabolismo , Lentivirus/genética , Proteínas de Neoplasias/genética , Interferencia de ARN , Receptores de Superficie Celular/genética , Feto , Células HEK293 , Humanos , TransfecciónRESUMEN
Aquaporin-1 (AQP1) and aquaporin-3 (AQP3) water channels expressed in the kidney play a critical role in the urine concentrating mechanism. Mice with AQP1 or AQP3 deletion have a urinary concentrating defect. To better characterize this defect, we studied the influence of an acute urea load (300 mumol ip) in conscious AQP1-null, AQP3-null, and wild-type mice. Urine was collected and assayed every 2 h, from 2 h before (baseline) to 8 h after the urea load. Mice of all genotypes excreted the urea load in approximately 4 h with the same time course. Interestingly, despite their low baseline, the AQP3-null mice raised their urine osmolality and urea concentration progressively after the urea load to values almost equal to those in wild-type mice at 8 h. In contrast, urine non-urea solute concentration did not change. Urine volume fell in the last 4 h to about one-fourth of basal values. AQP1-null mice increased their urine flow rate much more than AQP3-null mice and showed no change in urine osmolality and urea concentration. The urea load strongly upregulated urea transporter UT-A3 expression in all three genotypes. These observations show that the lack of AQP3 does not interfere with the ability of the kidney to concentrate urea but impairs its ability to concentrate other solutes. This solute-selective response could result from the capacity of AQP3 to transport not only water but also urea. The results suggest a novel role for AQP3 in non-urea solute concentration in the urine.