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1.
CNS Neurosci Ther ; 30(2): e14574, 2024 02.
Artigo em Inglês | MEDLINE | ID: mdl-38421088

RESUMO

RATIONALE: Numerous epidemiological studies have reported a link between low testosterone levels and an increased risk of cerebrovascular disease in men. However, there is ongoing controversy surrounding testosterone replacement therapy due to potential side effects. PBMT has been demonstrated to improve cerebrovascular function and promote testosterone synthesis in peripheral tissues. Despite this, the molecular mechanisms that could connect PBMT with testosterone and vascular function in the brain of photothrombosis (PT)-induced stroke rats remain largely unknown. METHODS: We measured behavioral performance, cerebral blood flow (CBF), vascular permeability, and the expression of vascular-associated and apoptotic proteins in PT-induced stroke rats treated with flutamide and seven consecutive days of PBM treatment (350 mW, 808 nM, 2 min/day). To gain further insights into the mechanism of PBM on testosterone synthesis, we used testosterone synthesis inhibitors to study their effects on bEND.3 cells. RESULTS: We showed that PT stroke caused a decrease in cerebrovascular testosterone concentration, which was significantly increased by 7-day PBMT (808 nm, 350 mW/cm2 , 42 J/cm2 ). Furthermore, PBMT significantly increased cerebral blood flow (CBF) and the expression of vascular-associated proteins, while inhibiting vascular permeability and reducing endothelial cell apoptosis. This ultimately mitigated behavioral deficits in PT stroke rats. Notably, treatment with the androgen receptor antagonist flutamide reversed the beneficial effects of PBMT. Cellular experiments confirmed that PBMT inhibited cell apoptosis and increased vascular-associated protein expression in brain endothelial cell line (bEnd.3) subjected to oxygen-glucose deprivation (OGD). However, these effects were inhibited by flutamide. Moreover, mechanistic studies revealed that PBMT-induced testosterone synthesis in bEnd.3 cells was partly mediated by 17ß-hydroxysteroid dehydrogenase 5 (17ß-HSD5). CONCLUSIONS: Our study provides evidence that PBMT attenuates cerebrovascular injury and behavioral deficits associated with testosterone/AR following ischemic stroke. Our findings suggest that PBMT may be a promising alternative approach for managing cerebrovascular diseases.


Assuntos
Terapia com Luz de Baixa Intensidade , Acidente Vascular Cerebral , Humanos , Masculino , Ratos , Camundongos , Animais , Testosterona/metabolismo , Androgênios/metabolismo , Receptores Androgênicos/metabolismo , Células Endoteliais/metabolismo , Flutamida/farmacologia , Flutamida/uso terapêutico , Flutamida/metabolismo , Acidente Vascular Cerebral/terapia
2.
Phytomedicine ; 116: 154882, 2023 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-37210961

RESUMO

BACKGROUND: Artemisia annua, a well-known traditional Chinese medicine, is the main source for production of artemisinin, an anti-malaria drug. A. annua is distributed globally, with great diversity of morphological characteristics and artemisinin contents. Diverse traits among A. annua populations impeded the stable production of artemisinin, which needs an efficient tool to identify strains and assess population genetic homogeneity. PURPOSE: In this study, ribosomal DNA (rDNA), were characterized for A. annua for strains identification and population genetic homogeneity assessment. METHODS: The ribosomal RNA (rRNA) genes were identified using cmscan and assembled using rDNA unit of LQ-9 as a reference. rDNA among Asteraceae species were compared performing with 45S rDNA. The rDNA copy number was calculated based on sequencing depth. The polymorphisms of rDNA sequences were identified with bam-readcount, and confirmed by Sanger sequencing and restriction enzyme experiment. The ITS2 amplicon sequencing was used to verify the stability of ITS2 haplotype analysis. RESULTS: Different from other Asteraceae species, 45S and 5S linked-type rDNA was only found in Artemisia genus. Rich polymorphisms of copy number and sequence of rDNA were identified in A. annua population. The haplotype composition of internal transcribed spacer 2 (ITS2) region which had moderate sequence polymorphism and relative short size was significantly different among A. annua strains. A population discrimination method was developed based on ITS2 haplotype analysis with high-throughput sequencing. CONCLUSION: This study provides comprehensive characteristics of rDNA and suggests that ITS2 haplotype analysis is ideal tool for A. annua strain identification and population genetic homogeneity assessment.


Assuntos
Artemisia annua , Artemisininas , Asteraceae , Artemisia annua/genética , DNA Ribossômico/genética , Medicina Tradicional Chinesa
3.
J Ethnopharmacol ; 293: 115328, 2022 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-35489660

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The dried pericarp of Citrus reticulata Blanco (CP) occupies an important position in the history of clinical applications of traditional Chinese medicine (TCM). In traditional use, CP is used to treat diseases related to the digestive, respiratory, and cardiovascular systems, as well as to regulate Qi and promote blood circulation throughout the body. In China, a special cultivar of CP named Guang Chen Pi (GCP) which is collected exclusively from Citrus reticulata Blanco's cultivars 'Chachi', is considered to be the best CP with high medicinal and dietary value. Modern pharmacology shows that CP has high effect on regulating metabolic disorders and cardiovascular systems diseases. Atherosclerosis (AS) is not only an inflammatory disease but also cardiovascular lipid metabolism disorder. Foam cells formation is the hallmark of AS. Several reports indicated that CP can mitigate the development of AS, but involved signaling pathway and its role in foam cell formation is unclear. Since the main components of GCP has protective effects in cardiovascular diseases, we evaluated its effect of inhibiting foam cell formation to support the traditional usage of GCP. AIM OF THE STUDY: The objective of this study aims to investigate the effects of GCP on suppressing RAW264.7 foam cell formation and anti-inflammatory in vitro. MATERIALS AND METHODS: To evaluate the anti-foam cell formation and anti-inflammatory activity of GCP, oxidized low-density lipoprotein (ox-LDL) induced RAW264.7 macrophages model was involved. Meantime, foam cell developing status was also closely monitored. RT-qPCR and Western blot were then applied to further investigate receptors in associated signaling pathways. RESULTS: GCP shown inhibitory effect on macrophage-derived foam cell formation in Oil Red O staining analysis, which was further confirmed by flow cytometry of Dil-ox-LDL staining and TG and TC analysis. The HDL-mediated cholesterol efflux was also promoted by GCP. Mechanistic studies showed that GCP significantly down-regulate SRA1 and CD36 protein expression, while significantly increasing the expression of PPARγ, LXRα, SRB1 and ABCG1. Also, GCP reduced ox-LDL-induced inflammatory factors level, and inhibited phosphorylation of p38 MAPK, ERK1/2, JNK1/2, NF-κB p65 and IKKα/ß. CONCLUSIONS: GCP exhibited anti-atherogenic ability by interfering RAW264.7 foam cell formation, through inhibiting lipid uptake and promoting HDL-mediated cholesterol. PPARγ-LXRα-ABCG1/SRB1 pathway and its anti-inflammatory effect may involve. This proposed anti-foam cell formation activity is expected to provide new insight on comprehensive utilization of GCP.


Assuntos
Aterosclerose , Citrus , Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Colesterol/metabolismo , Células Espumosas , Lipoproteínas LDL/metabolismo , Macrófagos , PPAR gama/metabolismo
4.
Food Chem ; 372: 131255, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-34627084

RESUMO

Tibet's native fruit resources have not been explored and remain underutilized due to geographical isolation. "Medog lemon" (ML), an ethno-species of wild citron, is an important functional food and dietary resource for the local people in southeastern Tibet. Herein comprehensive characterization of components of ML were firstly performed with an integrated strategy based on UHPLC-QE Orbitrap MS. A total of 196 components were characterized, among which 33 were detected from Citrus genus for the first time, and 55 were identified as potential new phytochemicals. The chemical comparison of ML with cultivated citron and fingered citron based on partial least squares-discriminant analysis model and univariate analysis revealed the distinct chemical composition of ML and in which more than 30 differentiated components were identified. The distinct morphological and chemical characters, as well as its good antioxidant properties, indicated ML as a potential new food resource that can be beneficial for human health.


Assuntos
Citrus , Frutas , China , Frutas/química , Humanos , Compostos Fitoquímicos , Extratos Vegetais/análise , Tibet
5.
PeerJ ; 9: e12325, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34721994

RESUMO

BACKGROUND: Amomum villosum Lour. is the plant that produces the famous traditional Chinese medicine Amomi Fructus. Frequent habitat destruction seriously threatens A. villosum germplasm resources. Genetic diversity is very important to the optimization of germplasm resources and population protection, but the range of inherited traits within A. villosum is unclear. In this study, we analyzed the genetic diversity and genetic structures of A. villosum populations in Guangdong and constructed a local reference DNA barcode library as a resource for conservation efforts. METHODS: DNA barcoding and Inter-Simple Sequence Repeat (ISSR) markers were used to investigate the population genetics of A. villosum. Five universal DNA barcodes were amplified and used in the construction of a DNA barcode reference library. Parameters including percentage of polymorphic sites (PPB), number of alleles (Na), effective number of alleles (Ne), Nei's gene diversity index (H), and Shannon's polymorphism information index (I) were calculated for the assessment of genetic diversity. Genetic structure was revealed by measuring Nei's gene differentiation coefficient (Gst), total population genetic diversity (Ht), intra-group genetic diversity (Hs), and gene flow (Nm). Analysis of molecular variance (AMOVA), Mantel tests, unweighted pair-group method with arithmetic mean (UPGMA) dendrogram, and principal co-ordinates (PCoA) analysis were used to elucidate the genetic differentiation and relationship among populations. RESULTS: A total of 531 sequences were obtained from the five DNA barcodes with no variable sites from any of the barcode sequences. A total of 66 ISSR bands were generated from A. villosum populations using the selected six ISSR primers; 56 bands, 84.85% for all the seven A. villosum populations were polymorphic. The A. villosum populations showed high genetic diversity (H = 0.3281, I = 0.4895), whereas the gene flow was weak (Nm = 0.6143). Gst (0.4487) and AMOVA analysis indicated that there is obvious genetic differentiation amongA. villosum populations and more genetic variations existed within each population. The genetic relationship of each population was relatively close as the genetic distances were between 0.0844 and 0.3347.

6.
Front Pharmacol ; 12: 695560, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34456722

RESUMO

The root of Reynoutria multiflora (Thunb.) Moldenke (syn: Polygonum multiflorum Thunb.) is a distinguished herb that has been popularly used in traditional Chinese medicine. The raw Reynoutria multiflora (RRM) should be processed by steaming before use, and the processing time is not specified in the processing specification. Our previous studies showed that the efficacy and toxicity of processed Reynoutria multiflora (PRM) at different processing times were inconsistent. A comprehensive identification method was established in this study to find a quality marker of raw Reynoutria multiflora (RRM) and processed Reynoutria multiflora (PRM) with different processing times. Metabolomics based on ultra-high-performance liquid chromatography tandem quadrupole/electrostatic field orbitrap high-resolution mass spectrometry (UHPLC-Q-Exactive plus orbitrap MS/MS) was used in this study. Using the CD.2 software processed database, multivariate statistical analysis methods coupled with cluster analysis and heatmap were implemented to distinguish between RRMs and PRMs with different processing times. The results showed that RRM and PRMs processed for 4, 8, 12, and 18 h cluster into group 1, and PRM processed for 24 and 32 h into group 2, indicating that it can effectively distinguish between the two groups and twenty potential markers, made the highest contributions to the observed chemical differences between two groups. Among them, tetrahydroxystilbene-O-hexoside-O-galloyl and sucrose can be used to identify PRM processed for 24 h. Therefore, the properties of RRM changed after 24 h of processing, and the quality markers were screened to distinguish RRM and PPM. It can also be used as an important control technology for the processing of RM, which has wide application prospects.

7.
J Agric Food Chem ; 68(31): 8483-8495, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32610017

RESUMO

The peel of Citrus reticulata 'Chachi' (GCP), which is highly valued in China for its health-promoting effects, is usually collected at different development stages to be processed into various functional foods. In the present work, a rapid method based on ultra-high-performance liquid chromatography Q Exactive Orbitrap mass spectrometry-incorporated untargeted and pseudotargeted metabolomics analysis was developed to investigate the chemical variations in GCP at different ripeness stages. Samples that originated from an individual tree were collected at immature, near mature, and mature stages. A total of 112 compounds were identified or tentatively identified, and flavonoids malonyl glycosides and polymethoxyfolavones glycosides were reported for the first time. Untargeted metabolomics analysis indicated the distinct chemical profiles and significant changes during ripeness stages. Then, a validated pseudotargeted metabolomics method based on parallel reaction monitoring was further applied with a wide coverage of targeted compounds. The GCP samples were found differing in the content variations of flavonoid aglycones, flavonoid O-/C-glycosides, polymethoxyfolavones, limonoids, alkaloids, and phenolic acid, which are important for phenotypic variations at different development stages. The present study is expected to provide new insight on comprehensive utilization of citrus peels at different ripeness stages.


Assuntos
Citrus/química , Frutas/crescimento & desenvolvimento , Metabolômica/métodos , Extratos Vegetais/química , China , Cromatografia Líquida de Alta Pressão , Citrus/crescimento & desenvolvimento , Citrus/metabolismo , Flavonoides/química , Flavonoides/metabolismo , Frutas/química , Frutas/metabolismo , Glicosídeos/química , Glicosídeos/metabolismo , Limoninas/química , Limoninas/metabolismo , Extratos Vegetais/metabolismo , Espectrometria de Massas em Tandem
8.
Front Pharmacol ; 11: 569144, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33408629

RESUMO

The root of Reynoutria multiflora (Thunb.) Moldenke (syn.: Polygonum multiflorum Thunb., HSW) is a distinguished herb that has been popularly used in traditional Chinese medicine (TCM). Evidence of its potential side effect on liver injury has accumulated and received much attention. The objective of this study was to profile the metabolic characteristics of lipids in injured liver of rats induced by HSW and to find out potential lipid biomarkers of toxic consequence. A lipopolysaccharide (LPS)-induced rat model of idiosyncratic drug-induced liver injury (IDILI) was constructed and evident liver injury caused by HSW was confirmed based on the combination of biochemical, morphological, and functional tests. A lipidomics method was developed for the first time to investigate the alteration of lipid metabolism in HSW-induced IDILI rat liver by using ultra-high-performance liquid chromatography/Q-exactive Orbitrap mass spectrometry coupled with multivariate analysis. A total of 202 characterized lipids, including phosphatidylcholine (PC), lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE), lysophosphatidylethanolamine (LPE), sphingomyelin (SM), phosphatidylinositol (PI), lysophosphatidylinositol (LPI), phosphatidylserine (PS), phosphoglycerols (PG), and ceramide (Cer), were compared among groups of LPS and LPS + HSW. A total of 14 out 26 LPC, 22 out of 47 PC, 19 out of 29 LPE, 16 out of 36 PE, and 10 out of 15 PI species were increased in HSW-treated rat liver, which indicated that HSW may cause liver damage via interfering the phospholipid metabolism. The present work may assist lipid biomarker development of HSW-induced DILI and it also provide new insights into the relationships between phospholipid perturbation and herbal-induced idiosyncratic DILI.

9.
RSC Adv ; 9(40): 23373-23381, 2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-35514483

RESUMO

CRP (Citri Reticulatae Pericarpium), a famous traditional Chinese medicine, has also been extensively used in foods and condiments in dietary practice for centuries. According to the Chinese Pharmacopeia (2015 edition) it contains two subtypes, Guangchenpi (GCP) and Chenpi (CP). GCP exclusively originates from the pericarp of Citrus reticulata 'Chachi' cultivar and it's generally believed that GCP has superior qualities compared with the other main cultivars (CP). In the present study, an integrated approach combining LC-QTOF MS-based untargeted metabolomics analysis and DNA barcoding molecular identification was conducted to study the genetic diversity and chemical differences between GCP and CP. A validated UPLC-QTOF MS metabolomics method was established to identify markers by using PCA and OPLS-DA models. 34 identified metabolites could be used as chemical markers to distinguish effectively between the two subtypes. Among them polymethoxyflavones (PMF) such as hexamethoxyflavone (nobiletin and natsudaidain), pentamethoxyflavone (tangeretin and sinensetin), and tetramethoxyflavone are the most influential markers. Support vector machines were employed to classify all the samples and these markers showed good prediction accuracy (100%). The results of DNA barcoding showed that the secondary structure of the ITS2 sequences were significantly different among GCP and other three cultivars. The study indicated the integrated method could be a powerful and reliable analytical tool for differentiating GCP from CP.

10.
Molecules ; 24(1)2018 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-30577553

RESUMO

Herb genomics and comparative genomics provide a global platform to explore the genetics and biology of herbs at the genome level. Panax ginseng C.A. Meyer is an important medicinal plant for a variety of bioactive chemical compounds of which the biosynthesis may involve transport of a wide range of substrates mediated by oligopeptide transporters (OPT). However, information about the OPT family in the plant kingdom is still limited. Only 17 and 18 OPT genes have been characterized for Oryza sativa and Arabidopsis thaliana, respectively. Additionally, few comprehensive studies incorporating the phylogeny, gene structure, paralogs evolution, expression profiling, and co-expression network between transcription factors and OPT genes have been reported for ginseng and other species. In the present study, we performed those analyses comprehensively with both online tools and standalone tools. As a result, we identified a total of 268 non-redundant OPT genes from 12 flowering plants of which 37 were from ginseng. These OPT genes were clustered into two distinct clades in which clade-specific motif compositions were considerably conservative. The distribution of OPT paralogs was indicative of segmental duplication and subsequent structural variation. Expression patterns based on two sources of RNA-Sequence datasets suggested that some OPT genes were expressed in both an organ-specific and tissue-specific manner and might be involved in the functional development of plants. Further co-expression analysis of OPT genes and transcription factors indicated 141 positive and 11 negative links, which shows potent regulators for OPT genes. Overall, the data obtained from our study contribute to a better understanding of the complexity of the OPT gene family in ginseng and other flowering plants. This genetic resource will help improve the interpretation on mechanisms of metabolism transportation and signal transduction during plant development for Panax ginseng.


Assuntos
Ginsenosídeos/química , Ginsenosídeos/metabolismo , Magnoliopsida/metabolismo , Panax/química , Fatores de Transcrição/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Genoma de Planta/genética , Magnoliopsida/genética , Filogenia , Fatores de Transcrição/genética
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