RESUMEN
This study aims to investigate the component variations and spatial distribution of ginsenosides in Panax quinquefolium roots during repeated steaming and drying. Ultra performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to identify the ginsenosides in the root extract. Matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) was employed to visualize the spatial distribution and spatiotemporal changes of prototype ginsenosides and metabolites in P. quinquefolium roots. The UPLC results showed that 90 ginsenosides were identified during the steaming process of the roots, and polar ginsenosides were converted into low polar or non-polar ginsenosides. The content of prototype ginsenosides decreased, while that of rare ginsenosides increased, which included 20(S/R)-ginsenoside Rg_3, 20(S/R)-ginsenoside Rh_2, and ginsenosides Rk_1, Rg_5, Rs_5, and Rs_4. MALDI-MSI results showed that ginsenosides were mainly distributed in the epidermis and phloem. As the steaming times increased, ginsenosides were transported to the xylem and medulla. This study provides fundamental information for revealing the changes of biological activity and pharmacological effect of P. quinquefolium roots that are caused by repeated steaming and drying and gives a reference for expanding the application scope of this herbal medicine.
Asunto(s)
Ginsenósidos , Panax , Ginsenósidos/análisis , Espectrometría de Masas en Tándem , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Panax/química , Cromatografía Líquida de Alta Presión/métodos , Raíces de Plantas/químicaRESUMEN
OBJECTIVE: Functional constipation (FC) is a common intestinal disease worldwide. Despite the presence of criteria such as Roman IV, there is no standardized diagnosis and treatment algorithm in Hong Kong that combines both Western and Chinese medicine approaches. This study integrates current effective and safe diagnosis and treatment methods for FC and provides a clear and scientific pathway for clinical professionals and patients. METHODS: A systematic search of the PubMed, Cochrane Library, and China National Knowledge Infrastructure databases was performed from their inception to June 30th, 2022, collecting the current evidence about the efficacious integrative management for FC. We organized a meeting of professionals in fields relevant to treatment and management of FC to develop a consensus agreement on clinical pathway process. RESULTS: We developed a clinical pathway for the treatment of FC based on the most recent published guidelines and consultation with experts. This pathway includes a hierarchy of recommendations for every step of the clinical process, including clinical intake, diagnostic examination, recommended labs, diagnostic flowchart, and guidance for selection of therapeutic drugs. CONCLUSION: This pathway establishes clinical standards for the diagnosis and treatment of FC using Chinese medicine and Western medicine; it will help to provide high-quality medical services in Hong Kong for patients with FC. Please cite this article as: Wei DJ, Li HJ, Lyu ZP, Lyu AP, Bian ZX, Zhong LL. A clinical pathway for integrative medicine in the treatment of functional constipation in Hong Kong, China. J Integr Med. 2023; 21(6): 550-560.
Asunto(s)
Medicina Integrativa , Humanos , Hong Kong , Vías Clínicas , China , Estreñimiento/diagnóstico , Estreñimiento/terapiaRESUMEN
BACKGROUND: Patients with post-stroke depression (PSD) usually experience anxiety, hopelessness, and insomnia, which have a negative impact on their daily activities and post-stroke rehabilitation. Acupuncture (AC), as a minimally invasive technique, has become a popular choice for improving depression symptoms. However, it is still unclear which therapy is associated with the best outcomes for PSD. In this review, we aimed to explore the impact of AC in alleviating symptoms of PSD and to evaluate the difference in effectiveness between AC combined with pharmacotherapies and various non-pharmacotherapies. METHODS: Six databases and three clinical trials registration platforms were searched from inception to March 2023. Randomized clinical trial comparing needle-based AC with pharmacotherapy, and other non-pharmacotherapy or invalid group were included. Two independent reviewers identified eligible studies, and collected data using a pre-made form. A Bayesian network meta-analysis was conducted to assess and compare different techniques using RStudio 3.6.0 with the package 'GEMTC' V.0.8.1. The primary outcome was the efficacy for PSD assessed by scales measuring depressive symptoms. The secondary outcomes were effectiveness for neurological function and the quality of life. The ranking probabilities for all treatment interventions was performed using the Surface Under the Cumulative Ranking curve (SUCRA). The risk of bias was assessed by using the Revised Cochrane Risk of Bias tool 2. RESULTS: Sixty-two studies, involving 5308 participants published from 2003 to 2022, were included. The results showed that compared with western medicine (WM) (defined as pharmacotherapy for PSD), AC alone or with repetitive transcranial magnetic stimulation (RTMS), Traditional Chinese medicine (TCM) alone or with WM, were superior for alleviating depression symptoms. Compared to Usual Care, AC alone or plus other therapies could significantly decrease scores on the Hamilton Depression Rating scale. According to result of SUCRA, AC plus RTMS had the highest probability of improving depressive symptoms with a probability of 49.43%. CONCLUSIONS: The results of this study indicate that AC alone or combined with other therapies appears to be effective in improving depression symptoms of stroke survivors. Moreover, in comparison to WM, AC alone or plus RTMS, TCM, TCM with WM, or WM, were more effective in improving depression symptoms of PSD. Also, AC with RTMS seems to be the most effective with the highest probability. REGISTRATION: This study was registered in the International Prospective Register of Systematic Reviews (PROSPERO) database in November 2020 and updated in July 2021. The registration number is CRD42020218752.
Asunto(s)
Terapia por Acupuntura , Accidente Cerebrovascular , Humanos , Depresión/etiología , Depresión/terapia , Metaanálisis en Red , Teorema de Bayes , Calidad de Vida , Terapia por Acupuntura/métodos , Accidente Cerebrovascular/complicaciones , Accidente Cerebrovascular/terapia , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
The difference of astragaloside â £ content and the expression of its biosynthesis related genes in imitating wild Astragalus mongolicus(IWA) and cultivated A.mongolicus(CA) under different growth years were systematically compared and analyzed.Then the key enzyme genes affected the difference of astragaloside â £ content in the above two A.mongolicus were screened.High-perfo-rmance liquid chromatography(HPLC)was used to determine the content of astragaloside â £ in A.mongolicusunderthe above two diffe-rent growth patterns.Based on the Illumina HiSeq and PacBio high-throughput sequencing platforms, thesecond-and third-generation transcriptome sequencing(RNA-Seq)databaseof the two A.mongolicuswas constructed.The related enzyme genes in the biosynthetic pathway of astragaloside â £ were screened and verified byquantitative reverse transcriptase polymerase chain reaction(RT-qPCR).The RNA-sequencing(RNA-Seq) and RT-qPCR data of each gene were subjected to correlation analysis and trend analysis.The results showed that the variation trend of astragaloside â £ contentby HPLC wasthe same as that of genes by RNA-Seq and RT-qPCR in 1-4 year IWA and 1-2 year CA.The trend level of astragaloside â £ contentwas lower in 2-year IWA than 1-year IWA.Compared with 2-year IWA, 3-year IWA had an upward trend, while 4-year IWA hada downward trend versus 3-year IWA.Additionally, 1-year CA had increased trendthan 2-year CA.However, the content of astragaloside â £ in 5-year IWA was higher than that of 6-year IWA, which wasinconsistent with the findings of RNA-Seq and RT-qPCR.This study preliminarily clarifiedthat the difference of astragaloside â £ contentin 1-4 year IWA and 1-2 year CA wasclosely related to the expression of the upstream and midstream genes(MVK, CMK, PMK, MVD, SS) in the biosynthetic pathway.The results facilitate the production and planting of Radix Astragali seu Hedysari.
Asunto(s)
Planta del Astrágalo , Saponinas , Triterpenos , Planta del Astrágalo/genética , Planta del Astrágalo/metabolismo , Astragalus propinquus/genética , Saponinas/análisis , Saponinas/genética , Triterpenos/análisisRESUMEN
OBJECTIVE: To observe effects of Tongluo Zhitong (, TLZT) gel preparation on p53, miR-502-5p, NF-κBp65 in synovial tissue of knee osteoarthritis (KOA), and to explore mechanism of TLZT gel preparation in treating KOA. METHODS: Thirthy-six Wistar rats aged 8 weeks and weighed 200 to 220 g (meaned 208 g) were randomly divided into normal group, model group and traditional Chinese medicine (TCM) group, 12 rats in each group. KOA model was established by modified Hulth method. After 4 weeks of modeling, TCM group treated with TLZT gel preparation for external use, 3 times daily for 2 weeks;normal group and model group were fed normally without intervention. After treatment, morphological changes of specimens in each group were observed, changes of miR-502-5p in synovial tissue were detected by qPCR, and contents of p53, NF-κBp65, IL-1ß, TNF-α, MMP-13 in synovial tissue were detected by qPCR and Western Blot respectively. RESULTS: (1)Morphological observation of specimens showed that the articular cartilage in model group was hyaline and uneven, the synovial membranes were hypertrophic and proliferative with a large number of inflammatory cells infiltrating, the joint fluid was thicker in texture;the articular cartilage in TCM group was more transparent and smooth, synovial hyperplasia was mild with a small amount of inflammatory cell infiltration, the texture of articular fluid was clear and sparse. (2) Compared with normal group, content of miR-502-5p of synovial tissue in model and TCM group were increased, mRNA and expression of p53 decreased, expression of NF-κBp65, IL-1ß, TNF-α, MMP-13 increased. (3)Compared with model group, content of miR-502-5p in synovial tissue of TCM group decreased (P<0.05), mRNA and protein expression of p53 increased (P<0.05), mRNA and protein expression of NF-κBp65, IL-1ß, TNF-α, MMP-13 decreased (P<0.05). CONCLUSION: Expression of p53, miR-502 -5p, NF -κBp65 in synovial tissue is closely related to synovial hyperplasia and inflammatory reaction, TLZT gel preparation may reduce proliferation and inflammatory reaction of KOA synovium by regulating the expression of p53, miR- 502-5p, NF-κBp65 in synovial tissues.
Asunto(s)
MicroARNs , Osteoartritis de la Rodilla , Animales , Medicamentos Herbarios Chinos , Ratas , Ratas Wistar , Membrana Sinovial , Proteína p53 Supresora de TumorRESUMEN
Objective: Schisandra sphenanthera and S. chinensis are the two important medicinal plants that have long been used under the names of "Nan-Wuweizi" and "Wuweizi", respectively. The misuse of "Nan-Wuweizi" and "Wuweizi" in herbal medical products calls for an accurate method to distinguish these herbs. Chloroplast (cp) genomes have been widely used in species delimitation and phylogeny due to their uniparental inheritance and lower substitution rates than that of the nuclear genomes. To develop more efficient DNA markers for distinguishing S. sphenanthera, S. chinensis, and the related species, we sequenced the cp genome of S. sphenanthera and compared it to that of S. chinensis. Methods: The cp genome of S. sphenanthera was sequenced at the Illumina HiSeq platform, and the reference-guided mapping of contigs was obtained with a de novo assembly procedure. Then, comparative analyses of the cp genomes of S. sphenanthera and S. chinensis were carried out. Results: The cp genome of S. sphenanthera was 146 853 bp in length and consisted of a large single copy (LSC) region of 95 627 bp, a small single copy (SSC) region of 18 292 bp, and a pair of inverted repeats (IR) of 16 467 bp. GC content was 39.6%. A total of 126 functional genes were predicted, of which 113 genes were unique, including 79 protein-coding genes, 30 transfer RNA (tRNA) genes, and four ribosomal RNA (rRNA) genes. Five tRNA, four protein-coding genes, and all rRNA were duplicated in the IR regions. There were 18 intron-containing genes, including six tRNA genes and 12 protein-coding genes. In addition, 45 SSRs were detected. The whole cp genome of S. sphenanthera was 123 bp longer than that of S. chinensis. A total of 474 SNPs and 97 InDels were identified. Five genetic regions with high levels of variation (Pi > 0.015), trnS-trnG, ccsA-ndhD, psbI-trnS, trnT-psbD and ndhF-rpl32 were revealed. Conclusion: We reported the cp genome of S. sphenanthera and revealed the SNPs and InDels between the cp genomes of S. sphenanthera and S. chinensis. This study shed light on the species identification and further phylogenetic study within the genus of Schisandra.
RESUMEN
Nanwuweizi( Schisandrae Sphenantherae Fructus) and Wuweizi( Schisandrae Chinensis Fructus) have long-term history of use as common traditional Chinese medicines since the Eastern Han Dynasty( AD.25-220 year).However their information are always confused in ancient literature because they were both used as " Wuweizi". Nanwuweizi and Wuweizi are faced with problems such as confused distribution of producing areas,unclear source plants and efficacy characteristics,which limit modern resource development and application. Based on ancient literatures of materia medica,this study conducted a systematic review from several aspects,i.e. the name,distribution of producing areas,source plants,efficacy characteristics and processing of the two medicines in ancient time. This study clarified five main aspects,as following,ancient production areas and corresponding modern distribution areas; source plants used for medicinal purposes in ancient time; application period and application scope; efficacy characteristics in clinical application;processing method. This study provides a reference for evaluating the quality and for their clinical application and reasonable development of Nanwuweizi and Wuweizi.
Asunto(s)
Medicamentos Herbarios Chinos/historia , Medicina de Hierbas/historia , Materia Medica , Schisandra , China , Historia Antigua , Medicina Tradicional China , Fitoterapia , Plantas MedicinalesRESUMEN
OBJECTIVE: To observe the therapeutic effect of acupuncture on blood pressure variability (BPV) in elderly patients with cerebral infarction complicated with essential hypertension. METHODS: Seventy-six elderly patients with cerebral infraction complicated with essential hypertension were randomized into an observation group and a control group, 38 cases in each group. Xingnao Kaiqiao acupuncture and nifedipin were given in the control group. On the basis treatment in the control group, the Huoxue Sanfeng and Tiaohe Ganpi acupuncture was applied at Renying (ST 9), Hegu (LI 4), Quchi (LI 11), Zusanli (ST 36) and Taichong (LR 3) in the observation group for 30 min. The treatment was given once a day, 5 times a week for 8 weeks. The 24-hour ambulatory blood pressure was monitored in the two groups. The changes of blood pressure and blood pressure variability were observed before and after 8 weeks of treatment, and the occurrence of adverse reactions during the treatment were recorded. RESULTS: The mean systolic blood pressure (SBP), diastolic blood pressure (DBP), daytime SBP, DBP, nighttime SBP and DBP were reduced in the two groups after 8 weeks of treatment (P<0.05), and the results in the observation group were lower than those in the control group (P<0.05). After 8 weeks of treatment, the daytime, nighttime, 24 h systolic blood pressure variability (SBPV) and diastolic blood pressure variability (DBPV) in the observation group were lower than those before treatment (P<0.05), there was no significant difference between before and after treatment in the control group (P>0.05). CONCLUSION: Huoxue Sanfeng and Tiaohe Ganpi acupuncture method can effectively control the blood pressure and blood pressure variability in the elderly patients with cerebral infraction complicated with essential hypertension.
Asunto(s)
Terapia por Acupuntura , Infarto Cerebral , Hipertensión , Anciano , Presión Sanguínea , Monitoreo Ambulatorio de la Presión Arterial , Infarto Cerebral/complicaciones , Humanos , Hipertensión/complicacionesRESUMEN
BACKGROUND: Alzheimer' disease (AD) is an ultimately fatal degenerative brain disorder that has an increasingly large burden on health and social care systems. There are only five drugs for AD on the market, and no new effective medicines have been discovered for many years. Chinese medicinal plants have been used to treat diseases for thousands of years, and screening herbal remedies is a way to develop new drugs. METHODS: We used molecular docking to screen 30,438 compounds from Traditional Chinese Medicine (TCM) against a comprehensive list of AD target proteins. TCM compounds in the top 0.5% of binding affinity scores for each target protein were selected as our research objects. Structural similarities between existing drugs from DrugBank database and selected TCM compounds as well as the druggability of our candidate compounds were studied. Finally, we searched the CNKI database to obtain studies on anti-AD Chinese plants from 2007 to 2017, and only clinical studies were included. RESULTS: A total of 1,476 compounds (top 0.5%) were selected as drug candidates. Most of these compounds are abundantly found in plants used for treating AD in China, especially the plants from two genera Panax and Morus. We classified the compounds by single target and multiple targets and analyzed the interactions between target proteins and compounds. Analysis of structural similarity revealed that 17 candidate anti-AD compounds were structurally identical to 14 existing approved drugs. Most of them have been reported to have a positive effect in AD. After filtering for compound druggability, we identified 11 anti-AD compounds with favorable properties, seven of which are found in anti-AD Chinese plants. Of 11 anti-AD compounds, four compounds 5,862, 5,863, 5,868, 5,869 have anti-inflammatory activity. The compound 28,814 mainly has immunoregulatory activity. The other six compounds have not yet been reported for any biology activity at present. DISCUSSION: Natural compounds from TCM provide a broad prospect for the screening of anti-AD drugs. In this work, we established networks to systematically study the connections among natural compounds, approved drugs, TCM plants and AD target proteins with the goal of identifying promising drug candidates. We hope that our study will facilitate in-depth research for the treatment of AD in Chinese medicine.
RESUMEN
AIM: To study the chemical constituents of the fruits of Illicium henryi. METHOD: Chromatographic separations on silica gel, Sephadex LH-20 gel and MCI gel were used to isolate the compounds. The structures were elucidated based on extensive spectroscopic data analyses. RESULTS: Seven compounds were obtained and their structures were identified as 10-benzoyl-cycloparvifloralone (1), cycloparvifloralone (2), 2α-hydroxycycloparviforalone (3), henrylactone B (4), merrillianone (5), henrylactone C (6) and 7, 14-ortholactone- 3-hydroxyfloridanolide (7). CONCLUSION: Compound 1 is a new sesquiterpene lactone. The tested compounds showed weak anti-HBV activities on HBV surface antigen (HBsAg) secretion and HBV e antigen (HBeAg) secretion using Hep G2.2.15 cell line.
Asunto(s)
Frutas/química , Illicium/química , Extractos Vegetales/química , Sesquiterpenos/aislamiento & purificación , Antivirales/química , Antivirales/aislamiento & purificación , Antivirales/farmacología , Células Hep G2 , Virus de la Hepatitis B/efectos de los fármacos , Humanos , Estructura Molecular , Extractos Vegetales/farmacología , Sesquiterpenos/química , Sesquiterpenos/farmacologíaRESUMEN
Two new monoterpenes, p-mentha-1(7),8-dien-2-O-ß-D-glucoside and trans-2,4-dihydroxy-2,4-dimethyl-trans-1-acetic acid γ-lactone were isolated from the fruits of Illicium lanceolatum along with trans-2,4-dihydroxy-2,4-dimethyl-cis-1-acetic acid γ-lactone, (1R,2R,4R)-8-p-menthen-1,2-diol, trans-sobrerol, (1S,2S,4R)-p-menthane-1,2,8-triol and (1S, 2S, 4R, 8R)-p-menthane-1,2,9-triol. The structures of the isolates were confirmed by spectroscopic analysis and they showed no inhibitory effects on the in vitro growth of microbial organisms (Escherichia coli, Staphyloccocus aureus, Bacillus subtilis) at less than 1.0 mg/mL.
Asunto(s)
Frutas/química , Illicium/química , Monoterpenos/química , Extractos Vegetales/química , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Bacillus subtilis/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Pruebas de Sensibilidad Microbiana , Modelos Químicos , Modelos Moleculares , Conformación Molecular , Peso Molecular , Monoterpenos/aislamiento & purificación , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Espectrometría de Masa por Ionización de Electrospray , Staphylococcus aureus/efectos de los fármacosRESUMEN
AIM: To elucidate the roles of receptor tyrosine kinases RET and VEGFR2 and the RAF/MEK/ERK signaling cascade in cancer treatment with sorafenib. METHODS: The cell lines A549, HeLa, and HepG2 were tested. The enzyme activity was examined under cell-free conditions using 384-well microplate assays. Cell proliferation was evaluated using the Invitrogen Alarmar Blue assay. Gene expression was analyzed using the Invitrogen SYBR Green expression assays with a sequence detection system. Protein expression analysis was performed using Western blotting. RESULTS: Sorafenib potently suppressed the activities of cRAF, VEGFR2, and RET with IC(50) values of 20.9, 4 and 0.4 nmol/L, respectively. Sorafenib inhibited cRAF, VEGFR2, and RET via non-ATP-competitive, ATP-competitive and mixed-type modes, respectively. In contrast, sorafenib exerted only moderate cytotoxic effects on the proliferation of the 3 cell lines. The IC(50) values for inhibition of A549, HeLa, and HepG2 cells were 8572, 4163, and 8338 nmol/L, respectively. In the 3 cell lines, sorafenib suppressed the cell proliferation mainly by blocking the MEK/ERK downstream pathway at the posttranscriptional level, which in turn regulated related gene expression via a feed-back mechanism. CONCLUSION: This study provides novel evidence that protein kinases RET and VEGFR2 play crucial roles in cancer treatment with sorafenib.
Asunto(s)
Antineoplásicos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Niacinamida/análogos & derivados , Compuestos de Fenilurea/farmacología , Proteínas Proto-Oncogénicas c-ret/antagonistas & inhibidores , Receptor 2 de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Antineoplásicos/uso terapéutico , Western Blotting , Técnicas de Cultivo de Célula , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Sistema de Señalización de MAP Quinasas/genética , Neoplasias/enzimología , Neoplasias/patología , Niacinamida/farmacología , Niacinamida/uso terapéutico , Compuestos de Fenilurea/uso terapéutico , Proteínas Proto-Oncogénicas c-ret/genética , Interferencia de ARN/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la Polimerasa , Sorafenib , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genéticaRESUMEN
Microorganism-mediated dissimilatory Fe (III) reduction is recognized as the dominant mechanism for Fe(III) reduction to Fe(II) in non-sulfidogenic anaerobic environments, but the microorganisms involved, especially in paddy soil, are still poorly understood. In this paper, an enrichment culture was conducted to study the phylogenetic diversity of Fe (III)-reducing bacteria in paddy soil, with acetate or hydrogen as the electron donor and with ferrihydrite or goethite as the electron acceptor, and by the methods of terminal-restriction fragment length polymorphism (T-RFLP) technology and 16S rRNA genes cloning and sequencing. No matter what the electron donor and electron acceptor were supplemented, the most abundant microorganisms were Geobacter and Clostridiales, and Rhodocyclaceae were also abundant, when acetate was supplemented as electron donor, which suggested that besides Geobacter, Clostridiales and Rhodocyclaceae could be also the important Fe(III)-reducing bacteria in paddy soil.
Asunto(s)
Bacterias/metabolismo , Biodiversidad , Compuestos Férricos/metabolismo , Oryza/crecimiento & desarrollo , Microbiología del Suelo , Bacterias/clasificación , Bacterias/crecimiento & desarrollo , Geobacter/metabolismo , Hierro/metabolismo , Oxidación-Reducción , Filogenia , Rhodocyclaceae/metabolismoRESUMEN
OBJECTIVE: To investigate the effects of Ginsenoside Rb1 (G-Rb1) on the oxidative damage and extracellular matrix accumulation in rat renal tubular epethelial cells induced by transforming growth factor-beta1 (TGF-beta1). METHODS: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, 10 ng/mL TGF-beta1-induced group, G-Rb1 treated groups in which rat renal tubular epethelial cells were treated with different concentration of G-Rb1 (10 ng/mL, 20 ng/mL, 40 ng/mL) after TGF-beta1 induction, G-Rb1 40 ng/mL group and 100 nmol/L DPI(diphenyleneiodonium, an inhibitor of NADPH oxidase) group. Intracellular reactive oxidative species (ROS) level was measured by flowcytometry. p47phox protein expression was assessed by immunohistochemistry and western blotting method. The expressions of collagen I (Col-I) and fibronectin(FN) gene were measured by real-time PCR analysis. The protein level of Col-I and FN were quantitatively detected by enzyme-linked immunosorbent assay. RESULTS: TGF-beta1 at 10 ng/mL significantly increased the intercellular ROS production and p47phox expression (P < 0.05). The levels of Col-I and FN were also significantly up-regulated with the stimulation of 10 ng/mL TGF-beta1 (P < 0.05). Compared to TGF-beta1-induced group, G-Rb1 and DPI depressed TGF-beta1-induced ROS production and p47phox overexpression. Meanhile, G-Rb1 and DPI decreased the levels of Col-I and FN. CONCLUSION: G-Rb1 could inhibit TGF-beta1 induced ROS production and decrease the levels of Col-I and FN in a dose-dependent manner. The mechanism might be partly related to the suppression of p47phox expression.
Asunto(s)
Matriz Extracelular/metabolismo , Ginsenósidos/farmacología , Túbulos Renales/metabolismo , NADPH Oxidasas/metabolismo , Factor de Crecimiento Transformador beta1/farmacología , Animales , Línea Celular , Medicamentos Herbarios Chinos/farmacología , Células Epiteliales/citología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Túbulos Renales/citología , Túbulos Renales/efectos de los fármacos , NADPH Oxidasas/genética , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The medicinal herb, Panax notoginseng, has been used for thousands of years in traditional Chinese medicine and possesses anti-fibrosis properties. Epithelial-myofibroblast transition (EMT) plays an important role in renal tubulointerstitial fibrosis. The present study was designed to examine whether ginsenoside Rg1, a major active component isolated from Panax notoginseng, has an ability to block this phenotypic transition in rat renal tubular epithelial cells (NRK-52E) induced by transforming growth factor-beta1 (TGF-beta1). The morphology of tubular epithelial-myofibroblast transition was observed through light microscope and transmission electron microscopy. alpha-SMA and E-cadherin are two markers of tubular epithelial-myofibroblast transition, their protein expressions were assessed by immunohistochemistry and western blot analysis. Gene expression of alpha-SMA as well as the two major extracellular matrix components collagen I and fibronectin was measured by real-time PCR analysis. Enzyme-linked immunosorbent assay was used to quantitatively detect collagen I and fibronectin in the supernatant. Our results revealed that ginsenoside Rg1 obviously blocked morphologic transformation in NRK-52E induced by TGF-beta1. Meanwhile, ginsenoside Rg1 inhibited the expression of alpha-SMA and the loss of E-cadherin, subsequently decreased the levels of collagen I and fibronectin in a dose-dependent manner. In addition, western blot analysis indicated that ginsenoside Rg1 inhibited the expression of P-ERK1/2 in NRK-52E induced by TGF-beta1. These results suggest that ginsenoside Rg1 can restrain the process of EMT maybe via suppressing the expression of P-ERK1/2 in vitro.
Asunto(s)
Células Epiteliales/efectos de los fármacos , Epitelio/patología , Fibroblastos/efectos de los fármacos , Ginsenósidos/farmacología , Túbulos Renales/citología , Panax notoginseng , Extractos Vegetales/farmacología , Actinas/metabolismo , Animales , Cadherinas/metabolismo , Diferenciación Celular/efectos de los fármacos , Línea Celular , Colágeno Tipo I/metabolismo , Fibronectinas/metabolismo , Fibrosis , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Panax notoginseng/química , Raíces de Plantas , ARN Mensajero/metabolismo , Ratas , Factor de Crecimiento Transformador beta1/efectos adversosRESUMEN
OBJECTIVE: To investigate the effects of ginsenoside R(g1) on the transdifferentiation of rat renal tubular epethelial cells induced by transforming growth factor-beta1, (TGF-beta1). METHOD: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, TGF-beta1-induced group and treated with ginsenoside R(g1) at different concentration (10, 20, 40 mg x L(-1)) group. The morphology of tubular epithelial-myofibroblast transdifferentiation induced by TGF-beta1 was observed through light microscope. alpha-SMA and E-cadherin protein expression were assessed by immunohistochemistry and western blot analyses. alpha-SMA, collagen I and and fibronectin gene expression were assessed by real-time quantitative chain reaction. Enzyme-linked immunosorbent assay was used to quantitatively detect collagen I and fibronectin in the supernatant. RESULT: 10 mg x L(-1) TGF-beta1 could induce the transdifferentiation of tubular epithelial myofibroblast, showing fibroblast-like in morphology, with significantly enhanced expression of alpha-SMA, depressed expression of E-cadherin and increased secretion of fibronectin and collagen I (P < 0.05). Compared to TGF-beta1-induced group, ginsenoside R(g1) partly abrogated the alpha-SMA expression and E-cadherin depression triggered by TGF-beta1 in tubular epithelial cells in a dose-dependent manner (P < 0.05). Meanhile, ginsenoside R(g1) blocked morphologic transformation of tubular epithelial cells and decreased levels of collagen I and fibronectin (P < 0.05). CONCLUSION: Ginsenoside R(g1) could inhibit TGF-beta1 induced the tubular epithelial-myofibroblast transdifferentiation and decreased levels of collagen I and fibronectin in NRK52E.