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1.
Carbohydr Polym ; 271: 118438, 2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34364577

RESUMEN

Phellodendron chinense Schneid. was widely used as a medicinal herb for the treatment of diabetic osteoporosis in China. In this study, an arabinogalactan, named as PPCP-1, was isolated from the bark of Phellodendron chinense Schneid., and purified by DEAE-cellulose DE52 and Sephacryl S-200 HR column chromatography. The structure of PPCP-1 was characterized as a repeating unit consisting of →3)-ß-d-Galp-(1→, →3,6)-ß-d-Galp-(1→, →5)-α-l-Araf-(1→, →4)-α-d-Glcp-(1→, →3)-α-d-Glcp-(1→, →4)-α-d-Manp-(1→ with branches of →5)-α-l-Araf-(1→, →3,5)-α-l-Araf-(1→ and terminal α-l-Araf. Pharmacologically, the oral administration of PPCP-1 preserved osteoporosis associated with hyperglycemia by inhibiting α-glucosidase activity, improving glucose tolerance, decreasing the accumulation of advanced glycation end products (AGEs), as well as down-regulating the expression of receptor for AGEs in tibias of streptozotocin-induced diabetic rats. Collectively, the present study suggested that the arabinogalactan PPCP-1 from Phellodendron chinense Schneid. might potentially be used as functional foods for bone health and/or developed for drug discovery for alleviating diabetic osteoporosis.


Asunto(s)
Conservadores de la Densidad Ósea/uso terapéutico , Diabetes Mellitus Experimental/tratamiento farmacológico , Galactanos/uso terapéutico , Hipoglucemiantes/uso terapéutico , Osteoporosis/prevención & control , Phellodendron/química , Animales , Conservadores de la Densidad Ósea/química , Conservadores de la Densidad Ósea/aislamiento & purificación , Diabetes Mellitus Experimental/complicaciones , Galactanos/química , Galactanos/aislamiento & purificación , Productos Finales de Glicación Avanzada/metabolismo , Inhibidores de Glicósido Hidrolasas/química , Inhibidores de Glicósido Hidrolasas/aislamiento & purificación , Inhibidores de Glicósido Hidrolasas/uso terapéutico , Hipoglucemiantes/química , Hipoglucemiantes/aislamiento & purificación , Lisina/análogos & derivados , Lisina/metabolismo , Masculino , Osteoporosis/etiología , Ratas Wistar , Receptor para Productos Finales de Glicación Avanzada/metabolismo
2.
Phytother Res ; 34(12): 3273-3286, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32603019

RESUMEN

Depression is partially caused by inflammation in central nervous system. This study investigated the ameliorative effects of phenol glycosides (PG) from Ligustrum lucidum Ait. (Oleaceae) on neuroinflammation and depressive-like behavior in mice hypothalamus as well as the molecular mechanism. Mice were administered with PG extract for 2 weeks prior to treatment with LPS. The mice treated with PG extract showed resistance to LPS-induced reduction in body weight and LPS-induced depressive-like behaviors shown by sucrose preference, tail suspension test, forced swimming test and open field test. LPS-induced activation of microglial cells and elevation in protein expression of inflammatory cytokines including IL-1ß, RANTES and MCP-1 in hypothalamus of mice were abrogated by pre-treatment with PG extract. This extract down-regulated expression of TLR4, MyD88, NLRP3, renin and angiotensin II and decreased proportional area of Iba-1+ microglias in hypothalamus. Pre-treatment with PG extract inhibited LPS-triggered activation of CaSR/Gα11 signaling, stimulated 1-OHase expression in hypothalamus, and enhanced circulating 1,25(OH)2 D3 level. Overall, pre-treatment with PG extract ameliorated LPS-induced depressive-like behaviors by repressing neuroinflammation in mice hypothalamus which was attributed to its suppression on activation of microglia and production of inflammatory cytokines via acting on TLR4 pathway, CaSR and RAS cascade associated with improving vitamin D metabolism.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Depresión/tratamiento farmacológico , Frutas/química , Glicósidos/uso terapéutico , Hipotálamo/efectos de los fármacos , Inflamación/tratamiento farmacológico , Ligustrum/química , Extractos Vegetales/química , Animales , Glicósidos/farmacología , Masculino , Ratones
3.
J Vis Exp ; (147)2019 05 09.
Artículo en Inglés | MEDLINE | ID: mdl-31132042

RESUMEN

Traditional Chinese herbal medicine plays a role as an alternative method in treating many diseases, such as postmenopausal osteoporosis (POP). Gushukang (GSK) granules, a marketed prescription in China, have bone-protective effects in treating POP. Before administration to the body, one standard preparation procedure is commonly required, which aims to promote the release of active constituents from raw herbs and enhance the pharmacological effects as well as therapeutic outcomes. This study proposes a detailed protocol for using GSK granules in in vivo and in vitro experimental assays. The authors first provide a detailed protocol to calculate the animal-appropriate dosages of granules for in vivo investigation: weighing, dissolving, storage, and administration. Second, this article describes protocols for micro-CT scanning and the measurement of bone parameters. Sample preparation, protocols for running the micro-CT machine and quantification of bone parameters were evaluated. Third, serum-containing GSK granules are prepared, and drug-containing serum is extracted for in vitro osteoclastogenesis and osteoblastogenesis. GSK granules were intragastrically administered twice per day to rats for three consecutive days. Blood was then collected, centrifuged, inactivated, and filtered. Finally, serum was diluted and used for performing osteoclastogenesis and osteoblastogenesis. The protocol described here can be considered a reference for pharmacological investigations of herbal prescription medicines, such as granules.


Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Animales , Resorción Ósea/diagnóstico por imagen , Resorción Ósea/tratamiento farmacológico , Resorción Ósea/patología , Medicamentos Herbarios Chinos/farmacología , Femenino , Humanos , Ratones Endogámicos C57BL , Osteoblastos/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Ovariectomía
4.
Am J Chin Med ; 47(2): 457-476, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30834778

RESUMEN

Ligustroflavone is one major compound contained in active fraction from Fructus Ligustri Lucidi (the fruit of Ligustrum lucidum), which could regulate parathyroid hormone (PTH) levels and improve calcium balance by acting on calcium-sensing receptors (CaSR). This study aimed to explore the potency of ligustroflavone as a CaSR antagonist and its protective effects against diabetic osteoporosis in mice. LF interacted well with the allosteric site of CaSR shown by molecular docking analysis, increased PTH release of primary parathyroid gland cells and suppressed extracellular calcium influx in HEK-293 cells. The serum level of PTH attained peak value at 2 h and maintained high during the period of 1 h and 3 h than that before treatment in mice after a single dose of LF. Treatment of diabetic mice with LF inhibited the decrease in calcium level of serum and bone and the enhancement in urinary calcium excretion as well as elevated circulating PTH levels. Trabecular bone mineral density and micro-architecture were markedly improved in diabetic mice upon to LF treatment for 8 weeks. LF reduced CaSR mRNA and protein expression in the kidneys of diabetic mice. Taken together, ligustroflavone could transiently increase PTH level and regulate calcium metabolism as well as prevent osteoporosis in diabetic mice, suggesting that ligustroflavone might be an effective antagonist on CaSR.


Asunto(s)
Apigenina/farmacología , Complicaciones de la Diabetes/complicaciones , Glicósidos/farmacología , Ligustrum/química , Osteoporosis/etiología , Osteoporosis/prevención & control , Receptores Sensibles al Calcio/antagonistas & inhibidores , Animales , Apigenina/administración & dosificación , Apigenina/aislamiento & purificación , Densidad Ósea/efectos de los fármacos , Calcio/metabolismo , Hueso Esponjoso/metabolismo , Células Cultivadas , Expresión Génica/efectos de los fármacos , Glicósidos/administración & dosificación , Glicósidos/aislamiento & purificación , Células HEK293 , Humanos , Riñón/metabolismo , Masculino , Ratones Endogámicos C57BL , Glándulas Paratiroides/citología , Glándulas Paratiroides/metabolismo , Hormona Paratiroidea/metabolismo , Receptores Sensibles al Calcio/genética , Receptores Sensibles al Calcio/metabolismo , Factores de Tiempo
5.
J Vis Exp ; (123)2017 05 31.
Artículo en Inglés | MEDLINE | ID: mdl-28605360

RESUMEN

Traditional herbal medicine, an alternative medicine in the clinical setting, has received increased attention in recent years. Before delivery to the body, an additional extraction procedure is commonly required to release the active constituents from raw herbs. Water decoction is a classical extraction procedure that is still broadly used in the clinical settings. Here, we propose a detailed protocol for er-xian decoction (EXD) in order to apply herbal decoctions to experimental studies. The calculation of an animal-appropriate dose is described, as well as the four main steps of EXD: soaking, water decoction, filtration, and concentration. In addition, serum-containing EXD is introduced to rats as a means of in vitro validation. Here, rats were orally administered EXD for three days. Blood samples were then collected, inactivated, centrifuged, and filtered. The serum, diluted with the culture medium, can be utilized to treat cells or tissues in vitro. For example, EXD was applied to both in vivo and in vitro studies and demonstrated that EXD enhances osteogenesis. This protocol can be used as a reference for the preparation and application of herbal medicines.


Asunto(s)
Medicamentos Herbarios Chinos , Medicina de Hierbas , Animales , Densidad Ósea/efectos de los fármacos , Femenino , Ratones , Osteogénesis/efectos de los fármacos , Fitoterapia , Ratas , Transcriptoma
6.
Artículo en Inglés | MEDLINE | ID: mdl-27563332

RESUMEN

CEP hypertrophy is one of the characteristics of intervertebral disc degeneration (IDD). LIG exerts a protective effect on IDD in animal model. The effect of LIG on CEP hypertrophy is further investigated in the present study. Cells were isolated from hypertrophic samples obtained from patients during vertebral fusion surgery. Cellular proliferation and the expression of type I collagen (Col I) and TGF-ß1 were tested. In the bipedal rats, the edges of the CEP and the sizes of noncartilaginous outgrowth, as well as the expression of osteogenic markers, Col1a, ALP, Runx2, and TGF-ß1, were detected. Within two passages, the condensed hypertrophic CEP cells exhibited osteogenic capacity by bony-like nodules and ALP positive staining, along with increased expression of Col I and TGF-ß1. LIG inhibited proliferation of CEP cells and downregulated the expression of Col I and TGF-ß1 in vitro. Furthermore, LIG attenuated CEP hypertrophy on the lumbar spine of bipedal rats by reducing Col1a, ALP, Runx2, and TGF-ß1 mRNA expression and TGF-ß1 distribution in vivo. We concluded LIG exerted a preventive effect on CEP hypertrophy via suppression of TGF-ß1 levels. This information could be used to develop alternative therapeutic methods to treat spinal CEP hypertrophy.

7.
Stem Cells Int ; 2016: 4079210, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27064269

RESUMEN

We studied the bone mesenchymal stem cells (bMSCs) and gene profiles regulated by Er-Xian Decoction (EXD), a traditional Chinese herbal formula widely used for postmenopausal osteoporosis treatment. Six-month-old female Imprinting Control Region mice that underwent ovariectomy were treated with EXD. After 3 months, bone mass was evaluated by µCT and histological and immunohistochemical detection. The self-renewal and differentiation capacities of bMSCs were evaluated by colony-forming unit-fibroblastic, colony-forming unit-adipocyte, and alkaline phosphatase staining. In addition, the expression of 26991 genes of bMSCs ex vivo at 2 weeks after EXD-treatment or of bMSCs in vitro after exposure to conditioned serum from EXD-treated rats was measured and analyzed using NimbleGen Gene Expression Profiling and Cluster and pathway analysis. EXD treatment increased bone mass, elevating osteocalcin protein levels in vivo and facilitating the self-renewal and osteoblastic differentiation of bMSCs ex vivo. EXD rescued several gene expressions that were dysregulated by OVX. These genes overlapped and their functions were involved in ten pathways between ex vivo and in vitro experiments. EXD exerts an osteogenic effect on bMSCs in OVX induced osteoporotic mice. Our results contribute to further study of its molecular mechanism and traditional use in the treatment of postmenopausal osteoporosis.

8.
Trials ; 16: 64, 2015 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-25885543

RESUMEN

BACKGROUND: High incidences of neck pain morbidity are challenging in various situations for populations based on their demographic, physiological and pathological characteristics. Chinese proprietary herbal medicines, as Complementary and Alternative Medicine (CAM) products, are usually developed from well-established and long-standing recipes formulated as tablets or capsules. However, good quantification and strict standardization are still needed for implementation of individualized therapies. The Qishe pill was developed and has been used clinically since 2009. The Qishe pill's personalized medicine should be documented and administered to various patients according to the ancient TCM system, a classification of personalized constitution types, established to determine predisposition and prognosis to diseases as well as therapy and life-style administration. Therefore, we describe the population pharmacokinetic profile of the Qishe pill and compare its metabolic rate in the three major constitution types (Qi-Deficiency, Yin-Deficiency and Blood-Stasis) to address major challenges to individualized standardized TCM. METHODS/DESIGN: Healthy subjects (N = 108) selected based on constitutional types will be assessed, and standardized pharmacokinetic protocol will be used for assessing demographic, physiological, and pathological data. Laboratory biomarkers will be evaluated and blood samples collected for pharmacokinetics(PK) analysis and second-generation gene sequencing. In single-dose administrations, subjects in each constitutional type cohort (N = 36) will be randomly divided into three groups to receive different Qishe pill doses (3.75, 7.5 and 15 grams). Multiomics, including next generation sequencing, metabolomics, and proteomics, will complement the Qishe pill's multilevel assessment, with cytochrome P450 genes as targets. In a comparison with the general population, a systematic population pharmacokinetic (PopPK) model for the Qishe pill will be established and verified. TRIAL REGISTRATION: This study is registered at ClinicalTrials.gov, NCT02294448 .15 November 2014.


Asunto(s)
Protocolos Clínicos , Medicina Tradicional China , Adulto , Estudios Cruzados , Voluntarios Sanos , Humanos , Modelos Biológicos , FN-kappa B/antagonistas & inhibidores , Medicina de Precisión , Qi
9.
Artículo en Inglés | MEDLINE | ID: mdl-24872832

RESUMEN

Most chronic low back pain is the result of degeneration of the lumbar intervertebral disc. Ligustrazine, an alkaloid from Chuanxiong, reportedly is able to relieve pain, suppress inflammation, and treat osteoarthritis and it has the protective effect on cartilage and chondrocytes. Therefore, we asked whether ligustrazine could reduce intervertebral disc degeneration. To determine the effect of ligustrazine on disc degeneration, we applied a rat model. The intervertebral disc degeneration of the rats was induced by prolonged upright posture. We found that pretreatment with ligustrazine for 1 month recovered the structural distortion of the degenerative disc; inhibited the expression of type X collagen, matrix metalloproteinase (MMP)-13, and MMP3; upregulated type II collagen; and decreased IL-1 ß , cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) expression. In conclusion, ligustrazine is a promising agent for treating lumbar intervertebral disc degeneration disease.

10.
PLoS One ; 7(11): e47482, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23144821

RESUMEN

Renal interstitial fibrosis is characterized by increased extracellular matrix (ECM) synthesis. Epithelial-mesenchymal transition (EMT) in kidneys is driven by regulated expression of fibrogenic cytokines such as transforming growth factor-beta (TGF-ß). Yam, or Dioscorea alata (DA) is an important herb in Chinese medicine widely used for the treatment of clinical diabetes mellitus. However, the fibrosis regulatory effect of DA is unclear. Thus, we examined TGF-ß signaling mechanisms against EMT in rat fibroblast cells (NRK-49F). The characterization of DA water-extracts used various methods; after inducing cellular fibrosis in NRK-49F cells by treatment with ß-hydroxybutyrate (ß-HB) (10 mM), we used Western blotting to examine the protein expression in the TGF-ß-related signal protein type I and type II TGF-ß receptors, Smads2 and Smad3 (Smad2/3), pSmad2 and Smad3 (pSmad2/3), Smads4, Smads7, and EMT markers. These markers included E-cadherin, alpha-smooth muscle actin (α-SMA), and matrix metalloproteinase-2 (MMP-2). Bioactive TGF-ß and fibronectin levels in the culture media were determined using ELISA. Expressions of fibronectin and Snail transcription factor, an EMT-regulatory transcription factor, were assessed by immunofluorescence staining. DA extract dose-dependently (50-200 µg/mL) suppressed ß-HB-induced expression of fibronectin in NRK-49F cells concomitantly with the inhibition of Smad2/3, pSmad2/3, and Smad4. By contrast, Smad7 expression was significantly increased. DA extract caused a decrease in α-SMA (α-smooth muscle actin) and MMP-2 levels, and an increase in E-cadherin expression. We propose that DA extract might act as a novel fibrosis antagonist, which acts partly by down regulating the TGF-ß/smad signaling pathway and modulating EMT expression.


Asunto(s)
Medicamentos Herbarios Chinos/uso terapéutico , Transición Epitelial-Mesenquimal/efectos de los fármacos , Enfermedades Renales/tratamiento farmacológico , Enfermedades Renales/patología , Riñón/patología , Proteínas Smad Reguladas por Receptores/metabolismo , Ácido 3-Hidroxibutírico/metabolismo , Animales , Cadherinas/metabolismo , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fibronectinas/metabolismo , Fibrosis/tratamiento farmacológico , Fibrosis/metabolismo , Fibrosis/patología , Riñón/citología , Riñón/efectos de los fármacos , Riñón/metabolismo , Enfermedades Renales/metabolismo , Ratas , Transducción de Señal/efectos de los fármacos , Factores de Transcripción de la Familia Snail , Factores de Transcripción/metabolismo , Factor de Crecimiento Transformador beta/metabolismo
11.
Pharmazie ; 66(10): 784-90, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22026161

RESUMEN

Ossification of posterior longitudinal ligament (OPLL) is a common disease in Asian countries. Osteoblast differentiation in posterior longitudinal ligamentous fibroblast is a pathologic basis of OPLL. Nowadays, an effective pharmacotherapy for OPLL is still hunted for. YQHYRJ Recipe (YQHYRJ) is designed based on traditional Chinese medicine (TCM) theories, and previous clinic trials reported its effect on relieving syndromes of cervical spondylopathy. To clarify the YQHYRJ effect of OPLL on a cellular level, we induced mice fibroblasts from posterior longitudinal ligaments to differentiate into osteoblasts by human recombinant BMP-2, and treated them with YQHYRJ and its three sub-compounds: YQ, HY and RJ. YQHYRJ and the sub-compounds reduced the increase of fibroblast proliferation, mineralization, type I collagen secretion induced by BMP-2 via MTT, alizarin red staining and immunochemical examination. Moreover, these agents inhibited BMP-2 induced upregulation of ossification-related genes ALP, Col I and OC as well as BMP signal molecules Smad1, Smad 5 and Runx2 mRNA expression. These results suggested YQHYRJ to be effective in inhibiting osteoblast differentiation induced by BMP-2 in fibroblasts from posterior longitudinal ligament. YQHYRJ might be a promising medicine for preventing OPLL disease.


Asunto(s)
Proteína Morfogenética Ósea 2/farmacología , Diferenciación Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Fibroblastos/efectos de los fármacos , Ligamentos Longitudinales/metabolismo , Osteoblastos/efectos de los fármacos , Fosfatasa Alcalina/metabolismo , Animales , Separación Celular , Cromatografía Líquida de Alta Presión , Colágeno Tipo I/metabolismo , Colorantes , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Inmunohistoquímica , Técnicas In Vitro , Ligamentos Longitudinales/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Microscopía Electrónica de Transmisión , Osificación del Ligamento Longitudinal Posterior/metabolismo , Osificación del Ligamento Longitudinal Posterior/patología , Osteocalcina/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína Smad1/biosíntesis , Proteína Smad5/biosíntesis , Sales de Tetrazolio , Tiazoles
12.
Electrophoresis ; 28(5): 871-5, 2007 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-17315152

RESUMEN

The perennial Bupleurum root is thick and woody and contains high levels of interfering compounds. Common protein extraction methods have proved refractory towards the isolation of proteins suitable for 2-DE, due to the presence of interfering compounds. A novel method for extracting proteins suitable for 2-DE was established to overcome these problems. The main characteristic of this protocol is the partitioning of the proteins into the aqueous (fraction A-2), chloroform and isoamyl alcohol phases (A-3), and the interphase (A-1). The proteins are then extracted from each of these phases. From A-1, 85% (extracted protein against total proteins) proteins could be extracted and purified. For fraction A-2, a novel phenol extraction step is employed for the extraction of proteins. Based on the well-resolved 2-DE patterns, our protein preparation is free of interfering compounds. Using these methods (A-1, A-2, and A-3-3), a total of 3662 (1526 + 1128 + 1008) spots could be separated, and a protein yield of about 1.41 mg per 1.0 g fresh root material was obtained. To our knowledge, this is the first time that a protocol for protein extraction from perennial Bupleurum root has been reported that gives reproducible results. The protocol is expected to be applicable to other recalcitrant plant tissues as well.


Asunto(s)
Bupleurum/química , Proteínas de Plantas/aislamiento & purificación , Raíces de Plantas/química , Cloroformo/química , Electroforesis en Gel Bidimensional , Pentanoles/química , Extractos Vegetales/química , Proteínas de Plantas/química , Agua/química
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