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1.
Article in English | MEDLINE | ID: mdl-38981775

ABSTRACT

Almost 16 % of the global population is affected by neurological disorders, including neurodegenerative and cerebral neuroimmune diseases, triggered by acute or chronic inflammation. Neuroinflammation is recognized as a common pathogenic mechanism in a wide array of neurological conditions including Alzheimer's disease, Parkinson's disease, postoperative cognitive dysfunction, stroke, traumatic brain injury, and multiple sclerosis. Inflammatory process in the central nervous system (CNS) can lead to neuronal damage and neuronal apoptosis, consequently exacerbating these diseases. Itaconate, an immunomodulatory metabolite from the tricarboxylic acid cycle, suppresses neuroinflammation and modulates the CNS immune response. Emerging human studies suggest that itaconate levels in plasma and cerebrospinal fluid may serve as biomarkers associated with inflammatory responses in neurological disorders. Preclinical studies have shown that itaconate and its highly cell-permeable derivatives are promising candidates for preventing and treating neuroinflammation-related neurological disorders. The underlying mechanism may involve the regulation of immune cells in the CNS and neuroinflammation-related signaling pathways and molecules including Nrf2/KEAP1 signaling pathway, reactive oxygen species, and NLRP3 inflammasome. Here, we introduce the metabolism and function of itaconate and the synthesis and development of its derivatives. We summarize the potential impact and therapeutic potential of itaconate and its derivatives on brain immune cells and the associated signaling pathways and molecules, based on preclinical evidence via various neurological disorder models. We also discuss the challenges and potential solutions for clinical translation to promote further research on itaconate and its derivatives for neuroinflammation-related neurological disorders.

3.
Cancer Med ; 12(20): 20482-20496, 2023 10.
Article in English | MEDLINE | ID: mdl-37795569

ABSTRACT

BACKGROUND: Ocular metastasis (OM) is a rare metastatic site of primary liver cancer (PLC). The purpose of this study was to establish a clinical predictive model of OM in PLC patients based on machine learning (ML). METHODS: We retrospectively collected the clinical data of 1540 PLC patients and divided it into a training set and an internal test set in a 7:3 proportion. PLC patients were divided into OM and non-ocular metastasis (NOM) groups, and univariate logistic regression analysis was performed between the two groups. The variables with univariate logistic analysis p < 0.05 were selected for the ML model. We constructed six ML models, which were internally verified by 10-fold cross-validation. The prediction performance of each ML model was evaluated by receiver operating characteristic curves (ROCs). We also constructed a web calculator based on the optimal performance ML model to personalize the risk probability for OM. RESULTS: Six variables were selected for the ML model. The extreme gradient boost (XGB) ML model achieved the optimal differential diagnosis ability, with an area under the curve (AUC) = 0.993, accuracy = 0.992, sensitivity = 0.998, and specificity = 0.984. Based on these results, an online web calculator was constructed by using the XGB ML model to help clinicians diagnose and treat the risk probability of OM in PLC patients. Finally, the Shapley additive explanations (SHAP) library was used to obtain the six most important risk factors for OM in PLC patients: CA125, ALP, AFP, TG, CA199, and CEA. CONCLUSION: We used the XGB model to establish a risk prediction model of OM in PLC patients. The predictive model can help identify PLC patients with a high risk of OM, provide early and personalized diagnosis and treatment, reduce the poor prognosis of OM patients, and improve the quality of life of PLC patients.


Subject(s)
Eye Neoplasms , Liver Neoplasms , Humans , Quality of Life , Retrospective Studies , Machine Learning , Risk Factors , Liver Neoplasms/diagnosis
4.
PLoS One ; 15(9): e0238179, 2020.
Article in English | MEDLINE | ID: mdl-32881902

ABSTRACT

Carotenoid cleavage dioxygenase (CCD), a key enzyme in carotenoid metabolism, cleaves carotenoids to form apo-carotenoids, which play a major role in plant growth and stress responses. CCD genes had not previously been systematically characterized in Brassica napus (rapeseed), an important oil crop worldwide. In this study, we identified 30 BnCCD genes and classified them into nine subgroups based on a phylogenetic analysis. We identified the chromosomal locations, gene structures, and cis-promoter elements of each of these genes and performed a selection pressure analysis to identify residues under selection. Furthermore, we determined the subcellular localization, physicochemical properties, and conserved protein motifs of the encoded proteins. All the CCD proteins contained a retinal pigment epithelial membrane protein (RPE65) domain. qRT-PCR analysis of expression of 20 representative BnCCD genes in 16 tissues of the B. napus cultivar Zhong Shuang 11 ('ZS11') revealed that members of the BnCCD gene family possess a broad range of expression patterns. This work lays the foundation for functional studies of the BnCCD gene family.


Subject(s)
Brassica napus/enzymology , Dioxygenases/genetics , Genome, Plant , Plant Proteins/genetics , Arabidopsis/enzymology , Brassica napus/genetics , Carotenoids/metabolism , Chromosome Mapping , Dioxygenases/classification , Dioxygenases/metabolism , Gene Expression Regulation, Plant , Multigene Family , Phylogeny , Plant Proteins/classification , Plant Proteins/metabolism , Promoter Regions, Genetic
5.
Eur J Pharmacol ; 882: 173309, 2020 Sep 05.
Article in English | MEDLINE | ID: mdl-32598952

ABSTRACT

Studies have shown that the ginsenoside Rg1 can improve depressive symptoms in vitro and in vivo. However, the efficacy of Rg1on the hippocampal astrocyte gap junctions in depression are unclear. We mainly aimed to explore the relationship between Rg1, hippocampal astrocyte gap junctions and depression. Using primary cultured astrocytes, corticosterone (CORT) was used to induce stress. CORT (100 µM) significantly reduced the survival rate in astrocytes, and this effect was prevented by additional Rg1 administration. Interestingly, the gap junction blocker carbenoxolone (CBX) was able to revert this Rg1 effect. In in vivo models, one group was exposed to chronic unpredictable stress (CUS) for 47 days, while another group was bilaterally injected with CBX (100 µM) into the hippocampal CA1 region. Rats treated with Rg1 (20 mg/kg) showed an improvement in the sucrose preference and the forced swimming test in both models, indicating an antidepressive activity of Rg1. The levels of astrocyte gap junction connexin 43 (Cx43) were detected by immunofluorescence (IF) and western blotting. The levels of glial fibrillary acidic protein (GFAP) were detected by IF. The gap junctions in the hippocampal CA1 area were evaluated using dye transfer and electron microscopy. The reduction in Cx43 expression, the decrease in the Cx43 to GFAP ratio, the shorter dye diffusion distance, and the abnormal ultrastructure of gap junctions in rats exposed to CUS were markedly alleviated by concomitant Rg1 treatment. Taken together, the ginsenoside Rg1 could improve depression-like behavior in rats induced by astrocyte gap junction dysfunction in the hippocampus.


Subject(s)
Antidepressive Agents/therapeutic use , Astrocytes/drug effects , Depression/drug therapy , Gap Junctions/drug effects , Ginsenosides/therapeutic use , Stress, Psychological/drug therapy , Animals , Animals, Newborn , Antidepressive Agents/pharmacology , Astrocytes/metabolism , Astrocytes/ultrastructure , Cell Survival/drug effects , Cells, Cultured , Connexin 43/metabolism , Depression/metabolism , Gap Junctions/metabolism , Gap Junctions/ultrastructure , Ginsenosides/pharmacology , Glial Fibrillary Acidic Protein/metabolism , Hippocampus/cytology , Male , Rats, Sprague-Dawley , Stress, Psychological/metabolism
6.
Toxicol Lett ; 324: 20-29, 2020 May 15.
Article in English | MEDLINE | ID: mdl-31987890

ABSTRACT

Similar to other types of neuronal degeneration, Parkinson's disease (PD) is characterized by the aggregation of a pathological protein, α-synuclein. The endoplasmic reticulum (ER) is the principal site of protein synthesis, quality control and degradation. Genetic mutants, environmental insults and other factors disturb ER balance and induce the accumulation of misfolded/unfolded proteins, which initiate ER stress and disturb normal cell function. ER stress perturbs Ca2+ homeostasis and initiates the activation of autophagy and inflammasomes, which have been identified as risk factors for the development of PD. However, the mechanisms by which ER stress contributes to the processed of PD pathogenesis and development remain unclear. This review summarizes current knowledge of ER stress and highlights the principal role of ER stress in PD pathogenesis which may help reveal novel sight to illustrate the pathomechanism of PD.


Subject(s)
Endoplasmic Reticulum Stress/physiology , Parkinson Disease/etiology , Activating Transcription Factor 6/physiology , Adaptation, Physiological , Animals , Autophagy , Calcium/metabolism , Endoribonucleases/physiology , Humans , Parkinson Disease/physiopathology , Protein Serine-Threonine Kinases/physiology , Unfolded Protein Response , X-Box Binding Protein 1/physiology , eIF-2 Kinase/physiology
7.
Medicine (Baltimore) ; 98(39): e17245, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31574838

ABSTRACT

RATIONALE: Over the past decade, although several new entities of renal tumors have emerged, a form of renal cell carcinoma (RCC) that morphologically resembles epithelial-myoepithelial carcinoma has not been reported thus far. Herein, we describe a case of an unusual renal tumor that remained unclassified under a current RCC subtype, and briefly present its morphologic, immunophenotypic, and genetic features. PATIENT CONCERNS: The patient was an 85-year-old man who presented with hematuria and flank pain. Imaging studies revealed a left renal mass without enlarged lymph nodes. There were no abnormal masses or nodules in other organs. DIAGNOSES: The patient underwent no other treatment except the left radical nephrectomy under a clinical diagnosis of invasive urothelial carcinoma and was discharged on the thirteenth day. Histologically, the renal tumor showed biphasic proliferation of epithelial (strongly cytokeratin-positive; P63, P40, and vimentin-negative) and myoepithelial (strongly vimentin-positive; focal P63 and P40-positive; and weakly cytokeratin-positive) cells arranged in a perivascular pseudorosette-like pattern. No mutations were detected in multiple gene tests. According to the pathological structure, the patient was diagnosed as primary epithelial myoepithelial carcinoma-like renal tumor. INTERVENTIONS: To the best of our knowledge, the present tumor has not been previously described, and thus, this variant has not been integrated into a known form of PCC. Therefore, we cannot diagnose this type of tumor with other types of kidney tumors. OUTCOMES: Three years after primary diagnosis, the patient died of multiple organ failure result from multiple distant metastases. LESSONS: We present the first case of carcinoma of the kidney with EMC-like features and a perivascular pseudorosette-like growth pattern. Clinicians should be aware of the features of this uncommon variant of RCC to avoid diagnostic delays or misdiagnosis and prevent unnecessary or inappropriate treatment.


Subject(s)
Carcinoma, Renal Cell/pathology , Kidney Neoplasms/pathology , Myoepithelioma/pathology , Aged, 80 and over , Carcinoma, Renal Cell/genetics , Fatal Outcome , Humans , Immunophenotyping , Kidney/pathology , Kidney Neoplasms/genetics , Male , Myoepithelioma/genetics , Rosette Formation
8.
Behav Brain Res ; 362: 288-298, 2019 04 19.
Article in English | MEDLINE | ID: mdl-30654121

ABSTRACT

Glucocorticoids are vital stress response hormones and facilitate stress coping. However, sustained glucocorticoid exposure is associated with negative effects on brain. The precise role of glucocorticoids in depression and anxiety remains unclear. In the present study, we found that rats exposed to chronic unpredictable stress (CUS) showed anxiety-like behavior but not depressive-like behavior in the absence of glucocorticoid production. It was interesting to find that the level of serotonin (5-HT) and the expression of tryptophan hydroxylase-2 (TPH2) were decreased after CUS in the hippocampus in sham rats, while adrenalectomy (ADX) prevented such decreases. In addition, the neurogenesis in hippocampus decreased in both sham and ADX rats after stress exposure. Furthermore, inhibition of mineralocorticoid receptor (MR) with spironolactone induced anxiety like behavior in sham rats but not ADX rats. The proliferation of cells was blocked by spironolactone. In conclusion, our results indicate that MR-dependent neurogenesis was closely related with anxiety-like behavior.


Subject(s)
Behavior, Animal/drug effects , Glucocorticoids/pharmacology , Receptors, Glucocorticoid/drug effects , Receptors, Mineralocorticoid/drug effects , Animals , Anxiety/drug therapy , Anxiety/metabolism , Depression/drug therapy , Depression/metabolism , Glucocorticoids/metabolism , Male , Neurogenesis/drug effects , Rats, Sprague-Dawley , Receptors, Glucocorticoid/metabolism , Receptors, Mineralocorticoid/metabolism , Serotonin/metabolism , Stress, Psychological/drug therapy , Stress, Psychological/metabolism , Tryptophan Hydroxylase/metabolism
9.
Biomed Pharmacother ; 92: 962-971, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28618657

ABSTRACT

BACKGROUND: Hypothalamic-pituitary-adrenal (HPA) axis hyperactivity is a well-established pathological feature of major depression, accompanied by the persistent increase of glucocorticoid level and the dysfunction of hypothalamic-pituitary-gonadal (HPG) axis. Ginsenoside Rg1 (Rg1) is one of the most active ingredients of Panax ginseng, which has various biological activity. OBJECTIVE: This study aimed to investigate the antidepressive effects of Rg1 and elucidate its impact on neuroendocrine system. METHODS: The antidepressive effects of Rg1 were first analysed in mice, and was further identified in the chronic-unpredictable-mild-stress (CUMS) model and the gonadectomized (GDX) model. The effects of Rg1 on depression-like behaviour were analysed by the forced swimming test (FST), tail suspension test (TST), sucrose preference test, and measurement of pentobarbital-induced sleep. The serum corticosterone and testosterone levels were detected by ELISA. The protein levels of glucocorticoid receptor (GR) and androgen receptor (AR) were analysed by western blot and immunohistochemistry analysis. RESULTS: Rg1 significantly decreased the immobility time of mice in FST and TST. Furthermore, Rg1 alleviated anhedonia and hopelessness, decreased serum corticosterone level, and increased serum testosterone level, and the GR protein level in the PFC and hippocampus of the CUMS-treated rats. Moreover, Rg1 improved sleep disruption, down-regulated the serum corticosterone level, and increased AR protein level in the PFC of the GDX-treated mice. CONCLUSION: Together, these studies suggest that Rg1 displayed antidepressant activity through the modulation of the HPA and the HPG axis. These findings provide new mechanism involved in the antidepressive effects of Rg1 and propose theoretical clues for clinical therapies.


Subject(s)
Antidepressive Agents/pharmacology , Behavior, Animal/drug effects , Depression/prevention & control , Ginsenosides/pharmacology , Hypothalamo-Hypophyseal System/drug effects , Testis/drug effects , Affect/drug effects , Animals , Corticosterone/blood , Depression/metabolism , Depression/physiopathology , Depression/psychology , Disease Models, Animal , Dose-Response Relationship, Drug , Food Preferences/drug effects , Hindlimb Suspension , Hypothalamo-Hypophyseal System/metabolism , Hypothalamo-Hypophyseal System/physiopathology , Male , Mice , Motor Activity/drug effects , Orchiectomy , Rats, Sprague-Dawley , Receptors, Androgen/metabolism , Receptors, Glucocorticoid/metabolism , Sleep/drug effects , Stress, Psychological/complications , Stress, Psychological/psychology , Swimming , Testis/metabolism , Testis/physiopathology , Testosterone/blood , Time Factors
10.
Acta Pharmacol Sin ; 38(10): 1317-1328, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28649132

ABSTRACT

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model remains the most commonly used animal model of Parkinson's disease (PD). There are three MPTP-treatment schemes: acute, subacute and chronic. Considering the advantages of the period and similarity to PD, the subacute model was often chosen to assess the validity of new candidates, but the changes caused by the subacute MPTP treatment and the appropriate positive control for this model remain to be further confirmed. The aim of this study was: to estimate the value of the subacute MPTP mouse model in aspects of behavioral performance, biochemical changes and pathological abnormalities, and to find effective positive drugs. Male C57BL/6 mice were injected with MPTP (30 mg·kg-1·d-1, ip) for 5 consecutive days. Three days before MPTP injection, the mice were orally administered selegiline (3 mg·kg-1·d-1), pramipexole (3 mg·kg-1·d-1), or medopar (100 mg·kg-1·d-1) for 18 days. Behavioral performance was assessed in the open field test, pole test and rotarod test. Neurotransmitters in the striatum were detected using HPLC. Protein levels were measured by Western blot. Pathological characteristics were examined by immunohistochemistry. Ultrastructure changes were observed by electron microscopy. The subacute MPTP treatment did not induce evident motor defects despite severe injuries in the dopaminergic system. Additionally, MPTP significantly increased the α-synuclein levels and the number of astrocytes in the striatum, and destroyed the blood-brain barrier (BBB) in the substantia nigra pars compacta. Both selegiline and pramipexole were able to protect the mice against MPTP injuries. We conclude that the subacute MPTP mouse model does not show visible motor defects; it is not enough to evaluate the validity of a candidate just based on behavioral examination, much attention should also be paid to the alterations in neurotransmitters, astrocytes, α-synuclein and the BBB. In addition, selegiline or pramipexole is a better choice than medopar as an effective positive control for the subacute MPTP model.


Subject(s)
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine/administration & dosage , Antiparkinson Agents/pharmacology , Disease Models, Animal , Parkinsonian Disorders/physiopathology , Animals , Astrocytes/drug effects , Astrocytes/metabolism , Behavior, Animal/drug effects , Benserazide/pharmacology , Benzothiazoles/pharmacology , Blood-Brain Barrier/metabolism , Chromatography, High Pressure Liquid/methods , Corpus Striatum/metabolism , Drug Combinations , Levodopa/pharmacology , Male , Mice , Mice, Inbred C57BL , Microscopy, Electron , Pramipexole , Selegiline/pharmacology , alpha-Synuclein/metabolism
11.
Acta Pharmacol Sin ; 37(12): 1525-1533, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27616576

ABSTRACT

AIM: Accumulation of α-synuclein (α-syn) in the brain is a characteristic of Parkinson's disease (PD). In this study, we investigated whether treatment with tunicamycin, an endoplasmic reticulum (ER) stress inducer, led to the accumulation of α-syn in PC12 cells, and where α-syn protein was accumulated, and finally, whether bibenzyl compound 20c, a novel compound isolated from Gastrodia elata (Tian ma), could alleviate the accumulation of α-syn and ER stress activation in tunicamycin-treated PC12 cells. METHODS: PC12 cells were treated with tunicamycin for different time (6 h, 12 h, 24 h, 48 h). Cell viability was determined by a MTT assay. Subcellular fractions of ER and mitochondria were extracted with the Tissue Endoplasmic reticulum Isolation Kit. The levels of α-syn protein and ER-stress-associated downstream chaperones were detected using Western blots and immunofluorescence. RESULTS: Treatment of PC12 cells with tunicamycin (0.5-10 µg/mL) dose-dependently increased the accumulation of α-syn monomer (19 kDa) and oligomer (55 kDa), and decreased the cell viability. Accumulation of the two forms of α-syn was observed in both the ER and mitochondria with increasing treatment time. Co-treatment with 20c (10-5 mol/L) significantly increased the viability of tunicamycin-treated cells, reduced the level of α-syn protein and suppressed ER stress activation in the cells, evidenced by the reductions in phosphorylation of eIF2α and expression of spliced ATF6 and XBP1. CONCLUSION: Tunicamycin treatment caused accumulation of α-syn monomer and oligomer in PC12 cells. Bibenzyl compound 20c reduces the accumulation of α-syn and inhibits the activation of ER stress, which protected PC12 cells against the toxicity induced by tunicamycin.


Subject(s)
Benzhydryl Compounds/pharmacology , Bibenzyls/pharmacology , Endoplasmic Reticulum Stress/drug effects , Gastrodia/chemistry , Phenols/pharmacology , Protective Agents/pharmacology , Tunicamycin/toxicity , Animals , PC12 Cells , Rats , alpha-Synuclein/metabolism
12.
Mol Cell Neurosci ; 71: 102-13, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26704905

ABSTRACT

Growing evidence indicates that GQ1b, one of the gangliosides members, contributes to synaptic transmission and synapse formation. Previous studies have shown that GQ1b could enhance depolarization induced neurotransmitter release, while the role of GQ1b in asynchronous release is still largely unknown. Here in our result, we found low concentration of GQ1b, but not GT1b or GD1b (which were generated from GQ1b by plasma membrane-associated sialidases), evoked asynchronous dopamine (DA) release from both clonal rat pheochromocytoma PC12 cells and rat striatal slices significantly. The release peaked at 2 min after GQ1b exposure, and lasted for more than 6 min. This effect was caused by the enhancement of intracellular Ca(2+) and the activation of Pyk2. Inhibition of Pyk2 by PF-431396 (a dual inhibitor of Pyk2 and FAK) or Pyk2 siRNA abolished DA release induced by GQ1b. Moreover, Pyk2 Y402, but not other tyrosine site, was phosphorylated at the peaking time. The mutant of Pyk2 Y402 (Pyk2-Y402F) was built to confirm the essential role of Y402 activation. Further studies revealed that activated Pyk2 stimulated ERK1/2 and p-38, while only the ERK1/2 activation was indispensable for GQ1b induced DA release, which interacted with Synapsin I directly and led to its phosphorylation, then depolymerization of F-actin, thus contributed to DA release. In conclusion, low concentration of GQ1b is able to enhance asynchronous DA release through Pyk2/ERK/Synapsin I/actin pathway. Our findings provide new insights into the role of GQ1b in neuronal communication, and implicate the potential application of GQ1b in neurological disorders.


Subject(s)
Dopamine/metabolism , Exocytosis , Focal Adhesion Kinase 2/metabolism , Gangliosides/metabolism , Actins/metabolism , Animals , Calcium/metabolism , Dopaminergic Neurons/metabolism , Focal Adhesion Kinase 2/genetics , Male , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Mutation , PC12 Cells , Protein Binding , Rats , Rats, Sprague-Dawley , Second Messenger Systems , Synapsins/metabolism
13.
Acta Pharmacol Sin ; 36(3): 311-22, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25640478

ABSTRACT

AIM: Protopanaxtriol (Ppt) is extracted from Panax ginseng Mayer. In the present study, we investigated whether Ppt could protect against 3-nitropropionic acid (3-NP)-induced oxidative stress in a rat model of Huntington's disease (HD) and explored the mechanisms of action. METHODS: Male SD rats were treated with 3-NP (20 mg/kg on d 1, and 15 mg/kg on d 2-5, ip). The rats received Ppt (5, 10, and 20 mg/kg, po) daily prior to 3-NP administration. Nimodipine (12 mg/kg, po) or N-acetyl cysteine (NAC, 100 mg/kg, po) was used as positive control drugs. The body weight and behavior were monitored within 5 d. Then the animals were sacrificed, neuronal damage in striatum was estimated using Nissl staining. Hsp70 expression was detected with immunohistochemistry. Reactive oxygen species (ROS) generation was measured using dihydroethidium (DHE) staining. The levels of components in the Nrf2 pathway were measured with immunohistochemistry and Western blotting. RESULTS: 3-NP resulted in a marked reduction in the body weight and locomotion activity accompanied by progressive striatal dysfunction. In striatum, 3-NP caused ROS generation mainly in neurons rather than in astrocytes and induced Hsp70 expression. Administration of Ppt significantly alleviated 3-NP-induced changes of body weight and behavior, decreased ROS production and restored antioxidant enzymes activities in striatum. Moreover, Ppt directly scavenged free radicals, increased Nrf2 entering nucleus, and the expression of its downstream products heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidase 1 (NQO1) in striatum. Similar effects were obtained with the positive control drugs nimodipine or NAC. CONCLUSION: Ppt exerts a protective action against 3-NP-induced oxidative stress in the rat model of HD, which is associated with its anti-oxidant activity.


Subject(s)
Antioxidants/pharmacology , Basal Ganglia/drug effects , Huntington Disease/prevention & control , Neurons/drug effects , Neuroprotective Agents/pharmacology , Nitro Compounds , Propionates , Sapogenins/pharmacology , Animals , Basal Ganglia/metabolism , Basal Ganglia/pathology , Basal Ganglia/physiopathology , Behavior, Animal/drug effects , Disease Models, Animal , Dose-Response Relationship, Drug , HSP70 Heat-Shock Proteins/metabolism , Heme Oxygenase (Decyclizing)/metabolism , Humans , Huntington Disease/chemically induced , Huntington Disease/metabolism , Huntington Disease/pathology , Huntington Disease/physiopathology , Male , Mitochondria/drug effects , Mitochondria/metabolism , Motor Activity/drug effects , NAD(P)H Dehydrogenase (Quinone)/metabolism , NF-E2-Related Factor 2/metabolism , Neurons/metabolism , Neurons/pathology , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Signal Transduction/drug effects , Time Factors , Weight Loss/drug effects
14.
Acta Pharmacol Sin ; 35(8): 1031-44, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24976156

ABSTRACT

AIM: To investigate the anti-fibrosis effects of ginsenoside Rg1 on alcohol- and CCl4-induced hepatic fibrosis in rats and to explore the mechanisms of the effects. METHODS: Rats were given 6% alcohol in water and injected with CCl4 (2 mL/kg, sc) twice a week for 8 weeks. Rg1 (10, 20 and 40 mg/kg per day, po) was administered in the last 2 weeks. Hepatic fibrosis was determined by measuring serum biochemical parameters, HE staining, Masson's trichromic staining, and hydroxyproline and α-SMA immunohistochemical staining of liver tissues. The activities of antioxidant enzymes, lipid peroxidation, and Nrf2 signaling pathway-related proteins (Nrf2, Ho-1 and Nqo1) in liver tissues were analyzed. Cultured hepatic stellate cells (HSCs) of rats were prepared for in vitro studies. RESULTS: In the alcohol- and CCl4-treated rats, Rg1 administration dose-dependently suppressed the marked increases of serum ALT, AST, LDH and ALP levels, inhibited liver inflammation and HSC activation and reduced liver fibrosis scores. Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues. Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes. Treatment of the cultured HSCs with Rg1 (1 µmol/L) induced Nrf2 translocation, and suppressed CCl4-induced cell proliferation, reversed CCl4- induced changes in MDA, GPX, PCIII and HA contents in the supernatant fluid and α-SMA expression in the cells. Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro. CONCLUSION: Rg1 exerts protective effects in a rat model of alcohol- and CCl4-induced hepatic fibrosis via promoting the nuclear translocation of Nrf2 and expression of antioxidant enzymes.


Subject(s)
Alcohol Drinking/adverse effects , Carbon Tetrachloride , Ginsenosides/therapeutic use , Liver Cirrhosis/drug therapy , Liver Cirrhosis/etiology , Liver/drug effects , NF-E2-Related Factor 2/metabolism , Animals , Cells, Cultured , Ginsenosides/isolation & purification , Hepatic Stellate Cells/drug effects , Hepatic Stellate Cells/metabolism , Hepatic Stellate Cells/pathology , Lipid Peroxidation/drug effects , Liver/pathology , Liver Cirrhosis/chemically induced , Liver Cirrhosis/pathology , Male , Panax/chemistry , Rats, Wistar , Signal Transduction/drug effects
15.
PLoS One ; 9(4): e94574, 2014.
Article in English | MEDLINE | ID: mdl-24718602

ABSTRACT

Protein tyrosine kinases, which are highly expressed in the central nervous system, are implicated in many neural processes. However, the relationship between protein tyrosine kinases and neurotransmitter release remains unknown. In this study, we found that ionomycin, a Ca²âº ionophore, concurrently induced asynchronous neurotransmitter release and phosphorylation of a non-receptor protein tyrosine kinase, proline-rich tyrosine kinase 2 (Pyk2), in clonal rat pheochromocytoma PC12 cells and cerebellar granule cells, whereas introduction of Pyk2 siRNA dramatically suppressed ionomycin-induced neurotransmitter release. Further study indicated that Tyr-402 (Y402) in Pyk2, instead of other tyrosine sites, underwent rapid phosphorylation after ionomycin induction in 1 min to 2 min. We demonstrated that the mutant of Pyk2 Y402 could abolish ionomycin-induced dopamine (DA) release by transfecting cells with recombinant Pyk2 and its mutants (Y402F, Y579F, Y580F, and Y881F). In addition, Src inhibition could prolong phosphorylation of Pyk2 Y402 and increase DA release. These findings suggested that Pyk2 was involved in ionomycin-induced neurotransmitter release through phosphorylation of Y402.


Subject(s)
Focal Adhesion Kinase 2/metabolism , Ionomycin/pharmacology , Neurotransmitter Agents/metabolism , Phosphotyrosine/metabolism , Animals , PC12 Cells , Phosphorylation/drug effects , RNA, Small Interfering/metabolism , Rats , src-Family Kinases/metabolism
17.
Chinese Journal of Oncology ; (12): 28-32, 2013.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-284245

ABSTRACT

<p><b>OBJECTIVE</b>To map the frequency and types of EGFR gene mutations present in lung cancer tissues. To evaluate the clinical applicability of a novel real-time double-loop probe PCR of which the ADx-EGFR kit is based, and to compare its performance with traditional Sanger DNA sequencing in the detection of somatic mutations of tumor genes.</p><p><b>METHODS</b>A total of 208 formalin-fixed paraffin-embedded (FFPE) tumor samples were tested. Genomic DNA of the tissue samples was extracted and purified, and subjected to both traditional PCR amplification, Sanger sequencing of EGFR gene in exon 18, 19, 20, 21, and ADx's EGFR mutation detection kit. The mutation rates for EGFR gene in exon 18, 19, 20, 21, as well as the frequency of each mutation detected by the two methods, were analyzed.</p><p><b>RESULTS</b>The traditional Sanger DNA sequencing technique was successfully performed in 196 out of 208 (94.2%) lung cancer samples, and 22 samples (11.2%) showed EGFR gene mutations. ADx-EGFR kit was successfully used in the lung cancers of all of the 208 cases (100.0%), and 40 samples (19.2%) showed mutations. In the lung cancer samples analyzed, mutations were mainly detected in the exon 19 and exon 21 L858R point mutation, i.e. 4.8% (10/208) and 11.6% (23/208) of total mutations, respectively, and the remaining mutations were rare.</p><p><b>CONCLUSIONS</b>The success rate of ADx-EGFR real-time PCR for formalin-fixed and paraffin-embedded tissues samples is significantly higher than that of Sanger sequencing (P < 0.01). There are significant differences between the two methods. ADx-EGFR real-time PCR shows a much higher successful detection rate and mutation rate of lung cancer tissues compared with that of Sanger sequencing. As a result, the real-time PCR with ADx-EGFR kit is proved to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing. It is an effective and reliable tool for clinical screening of somatic gene mutations in tumors.</p>


Subject(s)
Humans , DNA Mutational Analysis , Methods , Exons , Genes, erbB-1 , Lung Neoplasms , Genetics , Paraffin Embedding , Point Mutation , Real-Time Polymerase Chain Reaction , Methods
18.
Chinese Journal of Pathology ; (12): 757-761, 2010.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-295117

ABSTRACT

<p><b>OBJECTIVE</b>to map out the frequency and types of K-ras gene mutations present in colorectal and lung cancer patients; to evaluate the clinical applicability of a novel real-time double-loop probe PCR using the ADx-K-ras kit, and to compare its performance with the result by using traditional Sanger DNA sequencing in detection of somatic mutations of the tumor genes.</p><p><b>METHODS</b>a total of 827 formalin-fixed paraffin-embedded (FFPE) blocks including 583 from the colorectal and 244 from the lung cancer patients were assayed. Genomic DNA of the sample tissues was extracted, purified and subjected to PCR amplification of K-ras gene codon 12 and 13 and DNA sequencing was carried on using both the traditional Sanger sequencing method and the ADx's K-ras mutation detection kit, respectively. The mutation rates for K-ras gene at codon 12 and 13, and the mutation frequencies detected by using both methods were analyzed.</p><p><b>RESULTS</b>533 out of 583 (91.4%) colorectal cancer samples and 144 out of 244 lung cancer samples (59.0%) were detected using the traditional Sanger DNA sequencing technique, and 583 out of 583 (100.0%) colorectal plus 244 out of 244(100.0%) lung cancers were detected, respectively by using the ADx-K-ras kit. Of the 583 colorectal cancer samples, 192 (32.9%) showed mutations by using the ADx-K-ras kit in comparing with a result of 160 samples (27.4%) with K-ras gene mutation by using the traditional Sanger DNA sequencing technique. Of the 244 lung cancer samples, 26 (10.7%) showed K-ras gene mutations by using ADx-K-ras kit, while in 144 samples detected by using the traditional Sanger DNA sequencing technique, only 12 samples (8.3%) showed K-ras gene mutations. In colorectal cancer analyzed, GGT→GAT at codon 12 was the most common event with 35.1% (66/188) mutations, followed by GGC→GAC at codon 13 with 26.6% (50/188) and GGT→GTT at codon 12 with 18.6% (35/188), while GGT→GCT at codon12 was the most rare with only 1.6% (3/188) of the total mutation cases. In patients with lung cancer analyzed, GGT→GTT at codon 12 was the most common mutation, accounting for 40.9% (9/22), and GGT→GCT at codon 12 the most rare with only about 4.5% (1/22) of the total mutation cases.</p><p><b>CONCLUSIONS</b>K-ras gene mutations were present in colorectal cases, and significantly more frequent than that in lung cancer. There were significant statistical differences between the two methods. ADx-K-ras real-time PCR showed much higher successful detection rates and mutation ratios compared to Sanger sequencing. As a result, the real-time PCR with ADx-K-ras kit proves to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing. It is a effective and reliable tool for clinical screening of somatic gene mutations in tumors.</p>


Subject(s)
Humans , Colorectal Neoplasms , Genetics , Genes, ras , Genetics , Lung Neoplasms , Genetics , Mutation , Polymerase Chain Reaction , Methods , Sequence Analysis, DNA , Methods
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