Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 60
Filter
Add more filters

Country/Region as subject
Publication year range
1.
Cell Commun Signal ; 18(1): 66, 2020 04 20.
Article in English | MEDLINE | ID: mdl-32312253

ABSTRACT

BACKGROUND: Anti-nerve growth factor (NGF) monoclonal antibodies (anti-NGF mAbs) have been reported to significantly attenuate pain, but the mechanism involved has not been fully elucidated, and the serious adverse events associated with mAbs seriously limit their clinical use. This study further investigated the mechanism by which peripheral NGF is involved in neuropathic pain and found safe, natural compounds that target NGF to attenuate neuropathic pain. METHODS: Nociception was assessed by the Von Frey hair and Hargreaves' methods. Western-blotting, qPCR and immunofluorescence were used to detect the cell signaling pathway. RAW264.7 macrophages and RSC96 Schwann cells were cultured for in vitro evaluation. RESULTS: Intraplantar administration of anti-NGF mAbs suppressed the expression of phosphorylated transforming growth factor-ß-activated kinase 1 (TAK1) in the dorsal root ganglion (DRG) and sciatic nerve. Intraplantar administration of a TAK1 inhibitor attenuated CCI-induced neuropathic pain and suppressed the expression of phosphorylated mitogen-activated protein kinases (MAPKs) in the DRG and sciatic nerve. Perisciatic nerve administration of levo-corydalmine (l-CDL) on the operated side obviously attenuated CCI-induced neuropathic pain and suppressed the expression of mNGF and proNGF. In addition, l-CDL-induced antinociception was reversed by intraplantar administration of NGF. Further results indicated that l-CDL-induced suppression of phosphorylated TAK1, MAPKs, and p65 and expression of the proinflammatory cytokines TNF-α and IL-1ß in the DRG and sciatic nerve were all abolished by NGF. In addition, in vitro experiments indicated that l-CDL suppressed the secretion of NGF and proNGF in RAW264.7 macrophages and RSC96 Schwann cells, which was abolished by AP-1 and CREB agonists, respectively. CONCLUSIONS: This study showed NGF inhibition suppressed TAK1 in the periphery to attenuate CCI-induced neuropathic pain through inhibition of downstream MAPK and p65 signaling. The natural compound l-CDL inhibited NGF secretion by macrophages and Schwann cells and downstream TAK1-MAPK/NF-κB signaling in the periphery to attenuate CCI-induced neuropathic pain. Video abstract Proposed mechanisms underlying the effect of l-CDL in periphery of CCI rats. In CCI rats, macropahages and Schwann cells could secret NGF to act on the receptors in the periphery to activate TAK1-MAPK/NF-κB axis and promote the release of proinflammatory cytokines, including TNF-α and IL-1ß to promote neuropathic pain. l-CDL decreased the secretion of NGF through inhibiting AP-1 and CREB respectively in RAW264.7 and RSC96 Schwann cells to attenuate CCI-induced neuropathic pain by inhibiting the TAK1-p38 MAPK/NF-κB signaling pathway.


Subject(s)
Antibodies, Monoclonal , MAP Kinase Kinase Kinases , Nerve Growth Factor , Neuralgia/drug therapy , Plant Extracts , Animals , Antibodies, Monoclonal/pharmacology , Antibodies, Monoclonal/therapeutic use , Corydalis/chemistry , MAP Kinase Kinase Kinases/antagonists & inhibitors , MAP Kinase Kinase Kinases/metabolism , Mice , Nerve Growth Factor/antagonists & inhibitors , Nerve Growth Factor/immunology , Nerve Growth Factor/metabolism , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , RAW 264.7 Cells , Rats , Rats, Sprague-Dawley , Signal Transduction
2.
Analyst ; 145(10): 3620-3625, 2020 May 18.
Article in English | MEDLINE | ID: mdl-32338259

ABSTRACT

Early detection of acute kidney injury (AKI) is important, as early intervention and treatment can prevent further kidney injury and improve kidney health. Neutrophil gelatinase-associated lipocalin (NGAL) has emerged as the earliest and promising non-invasive biomarker of AKI in urine, and has been used as a new predictive biomarker of AKI in the bench-to-bedside journey. In this work, a nanocomplex composed of a polydopamine nanosphere (PDANS) and a fluorophore-labelled aptamer has been constructed for the detection of NGAL using a DNase I-assisted recycling amplification strategy. After the addition of NGAL, the fluorescence intensity increases linearly over the NGAL concentration range from 12.5 to 400 pg mL-1. The limit of detection of this strategy is found to be 6.25 pg mL-1, which is almost 5 times lower than that of the method that does not involve DNase I. The process can be completed within 1 h, indicating a fast fluorescence response. Furthermore, the method using the nanocomplex coupled with DNase I has been successfully utilized for the detection of NGAL in the urine from cisplatin-induced AKI and five-sixths nephrectomized mice, demonstrating its promising ability for the early prediction of AKI. This method also demonstrates the protective effect of the Huangkui capsule on AKI, and provides an effective way to screen potentially protective drugs for renal disease.


Subject(s)
Acute Kidney Injury/diagnosis , Aptamers, Nucleotide/metabolism , Deoxyribonuclease I/metabolism , Indoles/chemistry , Limit of Detection , Lipocalin-2/metabolism , Nanospheres/chemistry , Polymers/chemistry , Aptamers, Nucleotide/genetics , Cell Line , Humans , Nucleic Acid Amplification Techniques , Time Factors
3.
Molecules ; 23(9)2018 Sep 19.
Article in English | MEDLINE | ID: mdl-30235833

ABSTRACT

The interaction between proprotein convertase subtilisin/kexin type 9 (PCSK9) and the low-density lipoprotein receptor (LDLR) is a promising target for the treatment of hyperc-holesterolemia. In this study, a new method based on competitive affinity and tag detection was developed, which aimed to evaluate potent natural inhibitors preventing the interaction of PCSK9/LDLR directly. Herein, natural compounds with efficacy in the treatment of hypercholesterolemia were chosen to investigate their inhibitory activities on the PCSK9/LDLR interaction. Two of them, polydatin (1) and tetrahydroxydiphenylethylene-2-O-glucoside (2), were identified as potential inhibitors for the PCSK9/LDLR interaction and were proven to prevent PCSK9-mediated LDLR degradation in HepG2 cells. The results suggested that this strategy could be applied for evaluating potential bioactive compounds inhibiting the interaction of PCSK9/LDLR and this strategy could accelerate the discovery of new drug candidates for the treatment of PCSK9-mediated hypercholesterolemia.


Subject(s)
Biological Products/pharmacology , Glucosides/pharmacology , PCSK9 Inhibitors , Receptors, LDL/antagonists & inhibitors , Stilbenes/pharmacology , Cell Line, Tumor , Drugs, Chinese Herbal/pharmacology , Glucosides/chemistry , Glucosides/isolation & purification , Hep G2 Cells , Humans , Hypercholesterolemia/drug therapy , Hypercholesterolemia/genetics , Proprotein Convertase 9/metabolism , Protein Binding/drug effects , Receptors, LDL/metabolism
4.
Int J Cancer ; 141(4): 805-815, 2017 08 15.
Article in English | MEDLINE | ID: mdl-28500623

ABSTRACT

Bone cancer pain is a challenge for its not completely clarified mechanism and broad clinical morbidity. Therefore, novel and more effective drugs are urgent needed for improvement of patients' quality of life. Glutamate receptors have been associated with the development of the central sensitization of chronic pain. Inhibition of N-methyl-d-aspartate (NMDA) and metabotropic glutamate (mGlu) receptors can effectively attenuate bone cancer pain, respectively. Herein, our results indicated that levo-Corydalmine (l-CDL), a compound from Corydalis yanhusuo W.T. Wang, which has been used in traditional Chinese medicine for pain relief could effectively attenuate bone cancer pain induced by tibia bone cavity tumor cell implantation (TCI) through simultaneously inhibiting the NMDA and mGlu1/5 receptors in rat spinal cord without notable side effects. Both intragastric and intrathecal administration of l-CDL significantly alleviated the mechanical hypersensitivity induced by TCI in rats, and the analgesic effect of l-CDL could be reversed by intrathecal administration of NMDA receptor agonist NMDA and mGlu1/5 receptor agonist DHPG but not AMPA receptor agonist AMPA. l-CDL could also selectively suppress NMDA and DHPG induced rapid rise in Ca2+ oscillations in primary cultures neurons of spinal cord in vitro. The antinociception of l-CDL were partially mediated by the reduced phosphorylation of PKC γ and ERK1/2 in spinal cord of TCI rats in a NMDA and mGlu1/5 dependent manner. In conclusion, these results suggested that l-CDL attenuates TCI induced bone cancer pain through simultaneously inhibiting the NMDA and mGlu1/5 receptors and the downstream PKC γ, ERK1/2 signaling pathways in the spinal cord.


Subject(s)
Berberine/analogs & derivatives , Bone Neoplasms/complications , Cancer Pain/drug therapy , Receptors, Metabotropic Glutamate/antagonists & inhibitors , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Spinal Cord/enzymology , Animals , Berberine/administration & dosage , Berberine/pharmacology , Calcium/metabolism , Cancer Pain/enzymology , Gene Expression Regulation, Neoplastic/drug effects , Injections, Spinal , Male , Neoplasm Transplantation , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , Spinal Cord/drug effects , Spinal Cord/metabolism , Treatment Outcome
5.
Skeletal Radiol ; 46(11): 1597-1601, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28756568

ABSTRACT

We report the case of a 60-year-old man with a 12-day history of vomiting whenever he had a meal. Computed tomography revealed a calcified mass in the right cricoid cartilage with intraluminal and extraluminal extension. The patient underwent surgical resection of the trachea with end-to-end anastomosis. Pathological examination of the surgical specimen showed a low-grade chondrosarcoma. Eighteen months after surgery, the patient is alive and disease-free.


Subject(s)
Chondrosarcoma/diagnostic imaging , Chondrosarcoma/surgery , Cricoid Cartilage/diagnostic imaging , Cricoid Cartilage/surgery , Tomography, X-Ray Computed , Tracheal Neoplasms/diagnostic imaging , Tracheal Neoplasms/surgery , Anastomosis, Surgical , Diagnosis, Differential , Humans , Male , Middle Aged
6.
Biomed Chromatogr ; 29(3): 333-40, 2015 Mar.
Article in English | MEDLINE | ID: mdl-24961612

ABSTRACT

Neopanaxadiol (NPD), a major ginsenoside in Panax ginseng C. A. Meyer (Araliaceae), was reported to have neuroprotective effect. In this study, a method of ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/QTOF-MS) was developed and validated for quantitative analysis of NPD in tissues, urine and feces, using liquid-liquid extraction (LLE) to isolate NPD from different biological samples, and chromatographic separation was performed on an Agilent Zorbax Stable Bond C18 (2.1 × 50 mm, 1.8 µm) column with 0.1% formic acid in water and acetonitrile. All standard calibration curves were linear (all r(2) > 0.995) within the test range. After oral administration, NPD was extensively distributed to most of the tissues without long-term accumulation. The higher levels were observed in stomach and intestine, followed by kidney and liver. Approximately 64.56 ± 20.32% of administered dose in feces and 0.0233 ± 0.0356% in urine were found within 96 h, which indicated that the major elimination route was fecal excretion. This analytical method was applied to the study of NPD distribution and excretion in rats after oral intake for the first time. The results we found here are helpful for us to understand the pharmacological effects of NPD, as well as its toxicity.


Subject(s)
Chromatography, High Pressure Liquid/methods , Ginsenosides/analysis , Ginsenosides/pharmacokinetics , Mass Spectrometry/methods , Administration, Oral , Animals , Calibration , Feces , Ginsenosides/administration & dosage , Liquid-Liquid Extraction , Rats, Sprague-Dawley , Reproducibility of Results , Tissue Distribution
7.
Zhong Yao Cai ; 37(12): 2155-8, 2014 Dec.
Article in Zh | MEDLINE | ID: mdl-26080495

ABSTRACT

OBJECTIVE: To optimize the method for preparation of the insulin-like growth factor I (IGF-I ) in deer antler, and to determine the IGF-I in deer antler, heart and blood. METHODS: Ultrasonic extraction was used to extract IGF-I from different tissues of deer with ammonia-ammonium acetate buffer, followed by ultrafiltration and solid phase extraction to concentrate and purify the samples. At the same time, ethanol precipitation method was carried out in the purification of IGF-I ultrafiltratein deer antler, a parallel test proceeded and radio immune assay (RIA) was set to determine the IGF-I in deer antler, heart and blood. RESULTS: The IGF-I (60.8 ng/g) in deer antler by solid phase extraction was only existed in 30% methanol aqueous solution which was much higher than that (46.1 ng/g) by ethanol precipitation method. The quantities of IGF-I in deer antler, heart and blood were significantly different, it was 61.9 ng/g in antler and 21.9 ng/mL in blood, while there was no IGF-I tested in deer heart. CONCLUSION: Solid phase extraction is superior to ethanol precipitation method in preparing IGF-I in deer antler and it is clear that the IGF-I contained in deer antler is significantly higher than that in deer heart and blood, so it is the best choice to take IGF-I from deer antler.


Subject(s)
Antlers/chemistry , Biological Products/isolation & purification , Insulin-Like Growth Factor I/isolation & purification , Myocardium/chemistry , Animals , Blood , Deer
8.
J Agric Food Chem ; 72(13): 7043-7054, 2024 Apr 03.
Article in English | MEDLINE | ID: mdl-38509000

ABSTRACT

14-3-3ζ protein, the key target in the regulation and control of integrin ß3 outside-in signaling, is an attractive new strategy to inhibit thrombosis without affecting hemostasis. In this study, 4'-O-methylbavachalconeB (4-O-MB) in Psoraleae Fructus was identified as a 14-3-3ζ ligand with antithrombosis activity by target fishing combined with ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) analysis. The competitive inhibition analysis showed that 4-O-MB targeted 14-3-3ζ and blocked the 14-3-3ζ/integrin ß3 interaction with inhibition constant (Ki) values of 9.98 ± 0.22 µM. Molecular docking and amino acid mutation experiments confirmed that 4-O-MB specifically bound to 14-3-3ζ through LSY9 and SER28 to regulate the 14-3-3ζ/integrin ß3 interaction. Besides, 4-O-MB affected the integrin ß3 early outside-in signal by inhibiting AKT and c-Src phosphorylation. Meanwhile, 4-O-MB could inhibit ADP-, collagen-, or thrombin-induced platelet aggregation function but had no effect on platelet adhesion to collagen-coated surfaces in vivo. Administration of 4-O-MB could significantly inhibit thrombosis formation without disturbing hemostasis in mice. These findings provide new prospects for the antithrombotic effects of Psoraleae Fructus and the potential application of 4-O-MB as lead compounds in the therapy of thrombosis by targeting 14-3-3ζ.


Subject(s)
Platelet Aggregation , Thrombosis , Mice , Animals , Integrin beta3/genetics , Integrin beta3/chemistry , Integrin beta3/metabolism , 14-3-3 Proteins/genetics , 14-3-3 Proteins/metabolism , Platelet Glycoprotein GPIIb-IIIa Complex/genetics , Platelet Glycoprotein GPIIb-IIIa Complex/metabolism , Platelet Glycoprotein GPIIb-IIIa Complex/pharmacology , Molecular Docking Simulation , Thrombosis/drug therapy , Thrombosis/genetics , Thrombosis/metabolism , Collagen/metabolism , Blood Platelets/metabolism
9.
J Cancer ; 15(4): 1041-1052, 2024.
Article in English | MEDLINE | ID: mdl-38230224

ABSTRACT

Background: Dopamine receptors have been reported to be involved in pain, while the exact effects and mechanism in bone cancer pain have not been fully explored. Methods: Bone cancer pain model was created by implanting walker 256 mammary gland carcinoma into right tibia bone cavity. Primary cultured spinal neurons were used for in vitro evaluation. FLIPR, western-blot, immunofluorescence, and Co-IP were used to detect cell signaling pathway. Results: Our results indicated that spinal dopamine D1 receptor (D1DR) and spinal dopamine D2 receptor (D2DR) could form heteromers in TCI rats, and antagonizing spinal D1DR and D2DR reduced heteromers formation and alleviated TCI-induced bone cancer pain. Further results indicated that D1DR or D2DR antagonist induced antinociception in TCI rats could be reversed by D1DR, D2DR, and D1/D2DR heteromer agonists. And Gq, IP3, and PLC inhibitors also attenuated TCI-induced bone cancer pain. In vitro results indicated that D1DR or D2DR antagonist decreased the Ca2+ oscillations upregulated by D1DR, D2DR, and D1/D2DR heteromer agonists in activated primary cultured spinal neurons. Moreover, inhibition of D1/D2DR heteromers induced antinociception in TCI rats was partially mediated by the CaMKII and MAPKs pathway. In addition, a natural compound levo-Corydalmine (l-CDL), could inhibit D1/D2DR heteromers and attenuate bone cancer pain. Results: Inhibition of spinal D1/D2DR heteromers via l-CDL decreases excitability in spinal neurons, which might present new therapeutic strategy for bone cancer pain.

10.
Biomed Environ Sci ; 37(6): 639-646, 2024 Jun 20.
Article in English | MEDLINE | ID: mdl-38988114

ABSTRACT

Objective: To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Methods: We designed, developed, and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and detection. The precision of the liquid transfer and temperature control was tested. A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR). The entire process, from SARS-CoV-2 nucleic acid extraction to amplification, was evaluated. Results: The precision of the syringe transfer volume was 19.2 ± 1.9 µL (set value was 20), 32.2 ± 1.6 (set value was 30), and 57.2 ± 3.5 (set value was 60). Temperature control in the amplification tube was measured at 60.0 ± 0.0 °C (set value was 60) and 95.1 ± 0.2 °C (set value was 95) respectively. SARS-Cov-2 nucleic acid extraction yield through the device was 7.10 × 10 6 copies/mL, while a commercial kit yielded 2.98 × 10 6 copies/mL. The mean time to complete the entire assay, from SARS-CoV-2 nucleic acid extraction to amplification detection, was 36 min and 45 s. The detection limit for SARS-CoV-2 nucleic acid was 250 copies/mL. Conclusion: The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test (POCT).


Subject(s)
COVID-19 , Disposable Equipment , RNA, Viral , SARS-CoV-2 , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , COVID-19/virology , Humans , RNA, Viral/isolation & purification , RNA, Viral/analysis , COVID-19 Nucleic Acid Testing/instrumentation , COVID-19 Nucleic Acid Testing/methods , Nucleic Acid Amplification Techniques/instrumentation , Nucleic Acid Amplification Techniques/methods , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/instrumentation
11.
Bioorg Med Chem Lett ; 22(9): 3100-3, 2012 May 01.
Article in English | MEDLINE | ID: mdl-22483610

ABSTRACT

Lipase-catalyzed acylation of Guanfu alcohol-amine (GFAA) with vinyl acetate (VA) was performed in non-aqueous system for the preparation of Guanfu base G (GFG), a plant-originated alkaloid with significant antiarrhythmic activity. Among the eight lipases from different origins, Novozym 435 was found to be the best biocatalyst. The most suitable molecular sieve amount, substrate concentration, molar ratio of VA to GFAA, enzyme amount and reaction temperature were proved to be 40 mg/mL, 6 µmol/mL, 10:1, 2mg/mL and 50 °C, respectively. A maximum GFG yield of 37.4% was achieved under the selected conditions with methanol served as the optimal reaction medium. The structure of the acetylated product was elucidated by (1)H NMR and (13)C NMR analysis.


Subject(s)
Heterocyclic Compounds, 4 or More Rings/chemical synthesis , Lipase/metabolism , Acylation , Biocatalysis , Methanol , Methods , Molecular Structure , Vinyl Compounds/chemistry
12.
Appl Microbiol Biotechnol ; 93(6): 2357-64, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22189862

ABSTRACT

The microbial transformation of a series of tetrahydroprotoberberines (THPBs, 1-5) by Gliocladium deliquescens NRRL1086 was investigated. In this research, the novel glycosylation of tetrahydroberberrubine (1) was observed with fast rate and high regio- and enantio-selectivity. One pair of unique enantiomorphic alkaloidal glycosides T-1 and T-2 was isolated and their structures were elucidated unambiguously by HR-MS, CD, 1D and 2D NMR spectrum. It is interesting that different amounts of glucose in the potato broth medium could influence the ratio of T-1 and T-2; in the 1.5% glucose medium, the ratio was about 15:1 and the yield of the S-form product T-1 may reach the theoretical maximum yield of about 50% which could provide one practical method to prepare the enantiomerically pure product and one alternative resolution method of tetrahydroberberrubine. The preliminary enzymatic research by using sodium dodecyl sulfate (SDS) and imidazole as glycosyltransferase and glycosidase inhibitors revealed that glycosyltransferase may contribute to glycosylation process. This is the first successful approach to glycosylation of tetrahydroprotoberberines.


Subject(s)
Alkaloids/metabolism , Berberine Alkaloids/chemistry , Berberine Alkaloids/metabolism , Gliocladium/metabolism , Glycosides/metabolism , Biotransformation , Glycosylation , Molecular Structure , Stereoisomerism
13.
Int J Nanomedicine ; 17: 837-854, 2022.
Article in English | MEDLINE | ID: mdl-35228800

ABSTRACT

PURPOSE: In order to prepare a biomimetic nano-carrier which has inflammatory chemotaxis, homologous targeting and reduce immune clearance, for targeted chemotherapy of osteosarcoma, we fabricated the paclitaxel-loaded poly(lactic-co-glycolic) acid (PLGA) nanoparticles coated with 143B-RAW hybrid membrane (PTX-PLGA@[143B-RAW] NPs) and evaluate its anti-cancer efficacy in vitro and vivo. METHODS: PTX-PLGA@[143B-RAW] NPs were prepared by the ultrasonic method and were characterized by size, zeta potential, polymer dispersion index (PDI), Coomassie bright blue staining, transmission electron microscopy (TEM) and high performance liquid chromatography (HPLC). Cellular uptake, cell viability assay, flow cytometry and chemotactic effect of PTX-PLGA@[143B-RAW] NPs were evaluated in vitro. Biodistribution, anti-cancer therapeutic efficacy and safety of PTX-PLGA@[143B-RAW] NPs were evaluated in 143B osteosarcoma xenograft mice. RESULTS: The hybrid membrane successfully coated onto the surface of PLGA nanoparticles. PTX-PLGA@[143B-RAW] NPs had a drug loading capacity of 4.24 ± 0.02% and showed targeting ability to osteosarcoma. PTX-PLGA@[143B-RAW] NPs showed high cellular uptake and improved anti-cancer efficacy against 143B cells. More importantly, PTX-PLGA@[143B-RAW] NPs treatment suppressed tumor growth in tumor-bearing mice with minimal damage to normal tissues. CONCLUSION: PTX-PLGA@[143B-RAW] NPs could be used for targeted drug delivery and osteosarcoma therapy.


Subject(s)
Bone Neoplasms , Nanoparticles , Osteosarcoma , Animals , Biomimetics , Bone Neoplasms/drug therapy , Cell Line, Tumor , Cell Membrane , Drug Carriers/chemistry , Humans , Lactic Acid/chemistry , Mice , Nanoparticles/chemistry , Osteosarcoma/drug therapy , Paclitaxel , Polyglycolic Acid/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer , Tissue Distribution
14.
FASEB J ; 24(12): 4722-32, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20696856

ABSTRACT

Kava (Piper methysticum Foster, Piperaceae) organic solvent-extract has been used to treat mild to moderate anxiety, insomnia, and muscle fatigue in Western countries, leading to its emergence as one of the 10 best-selling herbal preparations. However, several reports of severe hepatotoxicity in kava consumers led the U.S. Food and Drug Administration and authorities in Europe to restrict sales of kava-containing products. Herein we demonstrate that flavokawain B (FKB), a chalcone from kava root, is a potent hepatocellular toxin, inducing cell death in HepG2 (LD(50)=15.3 ± 0.2 µM) and L-02 (LD(50)=32 µM) cells. Hepatocellular toxicity of FKB is mediated by induction of oxidative stress, depletion of reduced glutathione (GSH), inhibition of IKK activity leading to NF-κB transcriptional blockade, and constitutive TNF-α-independent activation of mitogen-activated protein kinase (MAPK) signaling pathways, namely, ERK, p38, and JNK. We further demonstrate by noninvasive bioluminescence imaging that oral consumption of FKB leads to inhibition of hepatic NF-κB transcriptional activity in vivo and severe liver damage. Surprisingly, replenishment with exogenous GSH normalizes both TNF-α-dependent NF-κB as well as MAPK signaling and rescues hepatocytes from FKB-induced death. Our data identify FKB as a potent GSH-sensitive hepatotoxin, levels of which should be specifically monitored and controlled in kava-containing herb products.


Subject(s)
Flavonoids/pharmacology , Glutathione/metabolism , Kava/chemistry , Mitogen-Activated Protein Kinases/metabolism , NF-kappa B/metabolism , Oxidative Stress/drug effects , Plant Roots/chemistry , Animals , Blotting, Western , Cell Line , Cell Survival/drug effects , Hep G2 Cells , Humans , I-kappa B Kinase/metabolism , Liver/drug effects , Liver/metabolism , Male , Mice , Mice, Inbred BALB C , Microscopy, Atomic Force , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effects , Tumor Necrosis Factor-alpha/pharmacology
15.
Exp Mol Med ; 53(2): 235-249, 2021 02.
Article in English | MEDLINE | ID: mdl-33558591

ABSTRACT

Dopamine D1 receptor (D1DR) and D2 receptor (D2DR) are closely associated with pain modulation, but their exact effects on neuropathic pain and the underlying mechanisms remain to be identified. Our research revealed that intrathecal administration of D1DR and D2DR antagonists inhibited D1-D2DR complex formation and ameliorated mechanical and thermal hypersensitivity in chronic constriction injury (CCI) rats. The D1-D2DR complex was formed in the rat spinal cord, and the antinociceptive effects of D1DR and D2DR antagonists could be reversed by D1DR, D2DR, and D1-D2DR agonists. Gαq, PLC, and IP3 inhibitors also alleviated CCI-induced neuropathic pain. D1DR, D2DR, and D1-D2DR complex agonists all increased the intracellular calcium concentration in primary cultured spinal neurons, and this increase could be reversed by D1DR, D2DR antagonists and Gαq, IP3, PLC inhibitors. D1DR and D2DR antagonists significantly reduced the expression of p-PKC γ, p-CaMKII, p-CREB, and p-MAPKs. Levo-corydalmine (l-CDL), a monomeric compound in Corydalis yanhusuo W.T. Wang, was found to obviously suppress the formation of the spinal D1-D2DR complex to alleviate neuropathic pain in CCI rats and to decrease the intracellular calcium concentration in spinal neurons. l-CDL-induced inhibition of p-PKC γ, p-MAPKs, p-CREB, and p-CaMKII was also reversed by D1DR, D2DR, and D1-D2DR complex agonists. In conclusion, these results indicate that D1DR and D2DR form a complex and in turn couple with the Gαq protein to increase neuronal excitability via PKC γ, CaMKII, MAPK, and CREB signaling in the spinal cords of CCI rats; thus, they may serve as potential drug targets for neuropathic pain therapy.


Subject(s)
Neuralgia/etiology , Neuralgia/metabolism , Receptors, Dopamine D1/metabolism , Receptors, Dopamine D2/metabolism , Sensory Receptor Cells/metabolism , Spinal Cord Injuries/etiology , Spinal Cord Injuries/metabolism , Animals , Behavior, Animal , Biomarkers , Calcium/metabolism , Cells, Cultured , Disease Models, Animal , Disease Susceptibility , Dopamine Antagonists/pharmacology , Dopamine D2 Receptor Antagonists/pharmacology , Male , Multiprotein Complexes/metabolism , Neuralgia/diagnosis , Pain Measurement , Phosphorylation , Protein Binding , Rats , Receptors, Dopamine D1/antagonists & inhibitors , Signal Transduction , Spinal Cord Injuries/complications , Spinal Cord Injuries/diagnosis
16.
J Adv Res ; 28: 139-148, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33364051

ABSTRACT

INTRODUCTION: Spinal N-methyl-D-aspartate receptor (NMDAR) is vital in chronic pain, while NMDAR antagonists have severe side effects. NMDAR has been reported to be controlled by G protein coupled receptors (GPCRs), which might present new therapeutic targets to attenuate chronic pain. Dopamine receptors which belong to GPCRs have been reported could modulate the NMDA-mediated currents, while their exact effects on NMDAR in chronic bone cancer pain have not been elucidated. OBJECTIVES: This study was aim to explore the effects and mechanisms of dopamine D1 receptor (D1DR) and D2 receptor (D2DR) on NMDAR in chronic bone cancer pain. METHODS: A model for bone cancer pain was established using intra-tibia bone cavity tumor cell implantation (TCI) of Walker 256 in rats. The nociception was assessed by Von Frey assay. A range of techniques including the fluorescent imaging plate reader, western blotting, and immunofluorescence were used to detect cell signaling pathways. Primary cultures of spinal neurons were used for in vitro evaluation. RESULTS: Both D1DR and D2DR antagonists decreased NMDA-induced upregulation of Ca2+ oscillations in primary culture spinal neurons. Additionally, D1DR/D2DR antagonists inhibited spinal Calcitonin Gene-Related Peptide (CGRP) and c-Fos expression and alleviated bone cancer pain induced by TCI which could both be reversed by NMDA. And D1DR/D2DR antagonists decreased p-NR1, p-NR2B, and Gαq protein, p-Src expression. Both Gαq protein and Src inhibitors attenuated TCI-induced bone cancer pain, which also be reversed by NMDA. The Gαq protein inhibitor decreased p-Src expression. In addition, D1DR/D2DR antagonists, Src, and Gαq inhibitors inhibited spinal mitogen-activated protein kinase (MAPK) expression in TCI rats, which could be reversed by NMDA. CONCLUSIONS: Spinal D1DR/D2DR inhibition eliminated NMDAR-mediated spinal neuron activation through Src kinase in a Gαq-protein-dependent manner to attenuate TCI-induced bone cancer pain, which might present a new therapeutic strategy for bone cancer pain.

17.
Biomed Microdevices ; 12(4): 655-63, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20300855

ABSTRACT

The use of ceramic nano-carriers containing anti-cancer drugs for targeted delivery that span both fundamental and applied research has attracted the interest of the scientific community. In this paper, a hydrophobic photodynamic therapy drug, hypocrellin A (HA), was successfully encapsulated in water-soluble amorphous silica nanocage (HANC) by an improved sol-gel method. These nanocages are of ultrasmall size, highly monodispersed, stable in aqueous suspension, and retain the optical properties of HA. Moreover, these nanocages can be effectively delivered, subsequently taken up by cancer cells and finally targeted to mitochondria. In addition, incubation time dependent photodynamic efficacy difference between HANC and HA was investigated for the first time. Especially, the nanocages, owning extremely high stable fluorescence comparing with free HA, also have potentials as efficient probes for optical biodiagnose in vitro. All these properties of HANC could possibly make it especially promising to be used as a bimodal reagent for photodynamic therapy and biodiagnose.


Subject(s)
Drug Carriers/metabolism , Mitochondria/metabolism , Perylene/analogs & derivatives , Photosensitizing Agents/chemistry , Quinones/chemistry , Silicon Dioxide/metabolism , Biological Transport , Drug Carriers/chemistry , HeLa Cells , Humans , Hydrogen Bonding , Indicators and Reagents/chemistry , Indicators and Reagents/metabolism , Intracellular Space/metabolism , Microscopy, Electron, Transmission , Mitochondria/drug effects , Mitochondria/radiation effects , Models, Molecular , Molecular Conformation , Perylene/chemistry , Perylene/pharmacology , Perylene/therapeutic use , Phenol , Photobleaching , Photochemotherapy , Photosensitizing Agents/pharmacology , Photosensitizing Agents/therapeutic use , Quinones/pharmacology , Quinones/therapeutic use , Silanes/chemistry , Silicon Dioxide/chemistry , Singlet Oxygen/metabolism , Spectroscopy, Fourier Transform Infrared
18.
Acta Pharmacol Sin ; 31(8): 984-9, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20686523

ABSTRACT

AIM: To generate a polyclonal antibody against sarsasapogenin and to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) method for the pharmacokinetic study of Sarsasapogenin in rats. METHODS: The antigen of sarsasapogenin was produced using an active ester method and subsequently used for raising polyclonal antibodies in rabbits. The specificity and sensitivity of the antibody were measured by IC-ELISA. Using the ELISA method, sarsasapogenin levels were measured in the serum of rats after an oral dose of 100 mg/kg. RESULTS: Polyclonal antibodies raised against sarsasapogenin-bovine serum albumin were generated and showed a high reactivity to sarsasapogenin. The antibodies exhibited minor cross-reactivity to ruscogenin (23%), diosgenin (22%), 25 (R, S) ruscogenin l-O-[beta-D-glucopyranosyl (1-->2)][beta-D-xylopyranosyl (1-->3)]-beta-D-fucopyranoside (26%) and no cross-reactivity to diammonium glycyrrhizinate and notoginseng R1. The detection range of sarsasapogenin by this ELISA method was approximately 2.4-760 ng/mL. The recovery rates of 10 ng/mL, 100 ng/mL, and 500 ng/mL were in the range of 91.0%-96.2% for intra-assay and 89.0%-92.0% for inter-assay. The coefficients of variation (CV%) for intra- and inter-assays at the three different sarsasapogenin levels were 3.1%-8.3% (n=6) and 6.0%-14.1% (n=6), respectively. CONCLUSION: The IC-ELISA method is a sensitive test for the determination of sarsasapogenin concentration in rat plasma and for pharmacokinetic (PK) studies.


Subject(s)
Antibodies/immunology , Enzyme-Linked Immunosorbent Assay/methods , Spirostans/blood , Animals , Cattle , Dose-Response Relationship, Drug , Male , Rabbits , Rats , Rats, Sprague-Dawley , Serum Albumin, Bovine/immunology
19.
Appl Microbiol Biotechnol ; 86(2): 491-7, 2010 Mar.
Article in English | MEDLINE | ID: mdl-19921181

ABSTRACT

The glycosylation of ruscogenin (1) by Gliocladium deliquescens NRRL 1086 was observed and gave a regioselectively glycosylated product identified as ruscogenin 1-O-beta-D-glucopyranoside (2) by infrared, mass spectrometry, and nuclear magnetic resonance spectra. Time-course studies indicated that it appeared to be favorable to accumulate 2 when ruscogenin was added to the 24-h-old stage II culture, and the yield of 2 was about 20.1% during 120 approximately 168 h. It was noted that additional carbohydrates could significantly increase glycoside formation and the yield of 2 even reached as high as 68% compared with the control 20.1%. The primary investigation about the characteristics of the enzyme resulted that the reaction was blocked by beta-glycosidase inhibitor imidazole, however, was enhanced remarkably by glycosyltransferase inhibitor sodium dodecyl sulfate. To our knowledge, this is the first reported case of producing steroidal saponin by microbial transformation, and G. deliquescens NRRL1086 would be a practical and highly efficient tool in producing natural ruscogenin monoside.


Subject(s)
Gliocladium/metabolism , Spirostans/metabolism , Carbon/metabolism , Glycosylation , Magnetic Resonance Spectroscopy , Mass Spectrometry , Stereoisomerism
20.
RSC Adv ; 10(19): 11257-11262, 2020 Mar 16.
Article in English | MEDLINE | ID: mdl-35495318

ABSTRACT

MicroRNA-21 (miRNA-21) is a significant biomarker which is closely related to some kinds of diseases, such as cancer, cardiovascular disease and kidney disease. Therefore, the detection of miRNA-21 is of great importance and can provide essential information for disease diagnosis. In this study, we report a facile, sensitive assay for miRNA-21 detection using personal glucose meters (PGM). Biotinylated DNA strand linked invertase (Inv) is conjugated on the surface of streptavidin-coated magnetic beads (MBs) to form a MBs-DNA-Inv complex. Target miRNA-21 in the detection system is captured by the MBs-DNA-Inv probe through DNA/RNA hybridization. The duplex-specific nuclease (DSN) enzyme specifically cleaves the DNA to recycle the target miRNA and release invertase, thereby triggering the dual signal amplification and ensuring high sensitivity. Besides, we establish a linear relationship between PGM and different concentrations of miRNA-21 in the range of 10 to 200 pM. The limit of detection is 1.8 pM, which is more sensitive than some of the previous reports. In addition, the biosensor exhibits excellent sequence selectivity and single-base mutation can be discriminated. Moreover, the expression of miRNA-21 is confirmed in urine from mice by our method, which is in good accordance with the qRT-PCR result. Therefore, a dependable, low-cost strategy for the detection of miRNA has been established and it meets the latest analytical demands for miRNA determination that is suitable for the public.

SELECTION OF CITATIONS
SEARCH DETAIL