ABSTRACT
Doxorubicin-based chemotherapy is a widely used first-line treatment for breast cancer, yet it is associated with various side effects, including splenic atrophy. However, the pathogenic mechanisms underlying doxorubicin-induced atrophy of the spleen remain unclear. This study investigates that doxorubicin treatment leads to splenic atrophy through several interconnected pathways involving histological changes, an inflammatory response, and apoptosis. Immunohistochemical and western blot analyses revealed reduced size of white and red pulp, decreased cellularity, amyloidosis, and fibrotic remodeling in the spleen following doxorubicin treatment. Additionally, increased secretion of pro-inflammatory cytokines was detected using an antibody array and enzyme-linked immunosorbent assay (ELISA), which triggers inflammation through the regulation of signal transducer and activator of transcription 3 (STAT3) and nuclear factor-kappa B (NF-κB) signaling pathways. Further analysis revealed that the loss of regulators and effectors of the oxidative defense system, including sirtuin (Sirt)3, Sirt5, superoxide dismutase (SOD)1, and SOD2, was implicated in the upstream regulation of caspase-dependent cellular apoptosis. These findings provide insights on the pathogenic mechanisms underlying doxorubicin-induced splenic atrophy and suggest that further investigation may be warranted to explore strategies for managing potential side effects in breast cancer patients treated with doxorubicin.
ABSTRACT
Clinical dose of doxorubicin (100 nM) induced cellular senescence and various secretory phenotypes in breast cancer and normal epithelial cells. Herein, we reported the detailed mechanism underlying ginsenoside Rh2-mediated NF-κB inhibition, and mitophagy promotion were evaluated by antibody array assay, western blotting analysis, and immunocytostaining. Ginsenoside Rh2 suppressed the protein levels of TRAF6, p62, phosphorylated IKK, and IκB, which consequently inactivated NF-κB activity. Rh2-mediated secretory phenotype was delineated by the suppressed IL-8 secretion. Senescent epithelial cells showed increased level of reactive oxygen species (ROS), which was significantly abrogated by Rh2, with upregulation on SIRT 3 and SIRT 5 and subsequent increase in SOD1 and SOD2. Rh2 remarkably favored mitophagy by the increased expressions of PINK1 and Parkin and decreased level of PGC-1α. A decreased secretion of IL-8 challenged by mitophagy inhibitor Mdivi-1 with an NF-κB luciferase system was confirmed. Importantly, secretory senescent epithelial cells promoted the breast cancer (MCF-7) proliferation while decreased the survival of normal epithelial cells demonstrated by co-culture system, which was remarkably alleviated by ginsenoside Rh2 treatment. These data included ginsenoside Rh2 regulated ROS and mitochondrial autophagy, which were in large part attributed to secretory phenotype of senescent breast epithelial cells induced by doxorubicin. These findings also suggested that ginsenoside Rh2 is a potential treatment candidate for the attenuation of aging related disease.
Subject(s)
Breast Neoplasms/chemically induced , Doxorubicin/adverse effects , Drugs, Chinese Herbal/therapeutic use , Ginsenosides/adverse effects , Mitochondria/metabolism , Autophagy , Breast Neoplasms/metabolism , Cell Culture Techniques , Cell Line, Tumor , Drugs, Chinese Herbal/pharmacology , Female , Humans , Oxidative StressABSTRACT
The administration of Dgalactose triggers brain aging by poorly understood mechanisms. It is generally recognized that Dgalactose induces oxidative stress or affects protein modifications via receptors for advanced glycated end products in a variety of species. In the present study, we aimed to investigate the involvement of astrocytes in Dgalactoseinduced brain aging in vitro. We found that Dgalactose treatment significantly suppressed cell viability and induced cellular senescence. In addition, as of the accumulation of senescent cells, we proposed that the senescenceassociated secretory phenotype (SASP) can stimulate agerelated pathologies and chemoresistance in brain. Consistently, senescent astrocytic CRT cells induced by Dgalactose exhibited increases in the levels of IL6 and IL8 via NFκB activation, which are major SASP components and inflammatory cytokines. Conditioned medium prepared from senescent astrocytic CRT cells significantly promoted the viability of brain tumor cells (U373MG and N2a). Importantly, conditioned medium greatly suppressed the cytotoxicity of U373MG cells induced by temozolomide, and reduced the protein expression levels of neuron marker neuronspecific class III ßtubulin, but markedly increased the levels of cMyc in N2a cells. Thus, our findings demonstrated that Dgalactose treatment might mimic brain aging, and that Dgalactose could contribute to brain inflammation and tumor progression through inducing the accumulation of senescentsecretory astrocytes.
Subject(s)
Astrocytes/metabolism , Astrocytoma/metabolism , Cellular Senescence , Drug Resistance, Neoplasm , Galactose/metabolism , Animals , Antineoplastic Agents/pharmacology , Astrocytes/drug effects , Astrocytoma/etiology , Cell Line, Tumor , Cell Survival/drug effects , Cells, Cultured , Cellular Senescence/drug effects , Cellular Senescence/genetics , Galactose/pharmacology , Humans , RatsABSTRACT
The acute exposure of trimethyltin (TMT) develops clinical syndrome characterized by amnesia, aggressive behavior, and complex seizures. This neurotoxicant selectively induces hippocampal neuronal injury and glial activation accompanied with resultant neuroinflammation. Here we report two candidates ginsenosides Rg3 and Rh2 as neuroprotection agents using a mouse model of TMT intoxication via a single injection (2 mg/kg) and primary neuronal culture systems. Four-week administration of Rg3 or Rh2 significantly reduced TMT-induced seizures and behavioral changes. Rg3 and Rh2 significantly attenuated the oxidative stress evidenced by improvement on antioxidant enzymes and neuronal loss and astrocytic activation in mouse brain. In primary cultures, TMT induced significant neuronal death after 24-h intoxication and vigorous secretion of inflammatory cytokines (IL-1α/ß, IL-6, TNF-α, and MCP-1) in astrocytes. Pretreatment with Rg3 or Rh2 not only reduced cell death but efficiently suppressed above mentioned inflammatory cytokines confirmed by antibody array test. The underlying protective mechanism by Rg3 and Rh2 was delineated through selective upregulation of PI3K/Akt and suppression of ERK activation. Intriguingly, Rg3 and Rh2 protected oligodendrocyte progenitor cells (O-2A) from TMT intoxication via promoting type 2 astrocytic differentiation without further inflammatory activation. Collectively, Rg3 and Rh2 interventions aimed at reducing oxidative stress and neuroinflammation neurotoxicity therefore are of therapeutic benefit in TMT-induced neurodegeneration.
Subject(s)
Encephalitis/prevention & control , Ginsenosides/pharmacology , Neurons/drug effects , Neuroprotective Agents/pharmacology , Animals , Antioxidants/pharmacology , Apoptosis/drug effects , Cell Death/drug effects , Cells, Cultured , Embryo, Mammalian , Encephalitis/chemically induced , Encephalitis/pathology , Neurons/physiology , Neuroprotection/drug effects , Neurotoxicity Syndromes/prevention & control , Oxidative Stress/drug effects , Rats , Rats, Sprague-Dawley , Trimethyltin CompoundsABSTRACT
Ginsenoside compound K (M1) is the active form of major ginsenosides deglycosylated by intestinal bacteria after oral administration. However, M1 was reported to selectively accumulate in liver and transform to fatty acid esters. Ester of M1 was not excreted by bile as M1 was, which means it was accumulated in the liver longer than M1. This study reported a synthetic method of M1-O, a mono-octyl ester of M1, and evaluated the anticancer property against murine H22 cell both in vitro and in vivo. As a result, both M1 and M1-O showed a dose-dependent manner in cytotoxicity assay in vitro. At lower dose of 12.5 µm, M1-O showed moderate detoxification. Instead, M1-O exhibited significantly higher inhibition in H22-bearing mice than M1. M1-O induced murine H22 tumor cellular apoptosis in caspase-dependent pathway given that pan-caspase inhibitor, Z-VAD-FMK, could reverse the cytotoxicity induced by M1-O. Additionally, pro- and anti-apoptosis proteins, Bcl-2 and Bax, altered and consequently induced increased expression of cleaved caspase-3. Interestingly, cyclophosphamide regimen significantly induced atrophy of spleen and thymus, main immune organs, while M1-O treatment greatly alleviated this atrophy. Collectively, we propose M1-O as a candidate for live cancer treatment.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Ginsenosides/chemistry , Amino Acid Chloromethyl Ketones/pharmacology , Animals , Antineoplastic Agents, Phytogenic/chemical synthesis , Antineoplastic Agents, Phytogenic/therapeutic use , Apoptosis/drug effects , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Caspase 3/chemistry , Caspase 3/metabolism , Cell Line, Tumor , Ginsenosides/chemical synthesis , Ginsenosides/pharmacology , Ginsenosides/therapeutic use , Liver Neoplasms/drug therapy , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Mice , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolismABSTRACT
Trimethyltin (TMT) is a potent neurotoxicant that affects various regions within the central nervous system, including the neocortex, cerebellum, and hippocampus. In the present study, ginsenoside Rd was investigated as a candidate neuroprotective agent in a primary hippocampal neuron culture and mouse models. TMT induced neurotoxicity in a seven-day primary hippocampal neuron culture in a dose-dependent manner (2.5-10 µM). However, pre-treatment with 20 µg/ml ginsenoside Rd for 24 h reversed the toxic action. ICR mice were administered a single injection of 2 mg/kg body weight TMT. Apparent tremor seizure and impaired passive avoidance tests demonstrated significant differences when compared with a saline treated control group. Nissl staining was performed to evaluate the neuronal loss in the hippocampus. In addition, immunostaining of glial fibrillary acidic protein characterized the features of astroglial activation. These results demonstrated that TMT markedly induced Cornu Ammonis 1 subregion neuronal loss and reactive astrocytes in the hippocampus, indicating disrupted hippocampal function. Notably, ginsenoside Rd attenuated the tremor seizures and cognitive decline in behavioral tests. Additionally, significantly reduced neuronal loss (P=0.018) and active astroglials (P=0.003) were observed in the ginsenoside Rd treated group. Ginsenoside Rd prevented TMT-induced cell apoptosis via regulation of B-cell lymphoma 2 (Bcl-2), bcl-2-like protein 4 and caspase-3. These results demonstrate that ginsenoside may be developed as a neuroprotective agent to prevent TMT-induced neurotoxicity.
ABSTRACT
Hydrogen peroxide was applied for promoting sporulation of Esteya vermicola and response surface methodology was used to optimize the effect of processing parameters on sporulation. Three variables were concentration (X 1), treatment time (X 2), and carbon-to-nitrogen ratio (X 3). The results indicated that X 1 and X 2 and the quadratic term of X 1 had significant effect on the sporulation, followed by the significant interaction effects between X 1 and X 2. The optimal conditions of promoting sporulation were as follows: hydrogen peroxide concentration 1.65 mM, treatment time 9.40 min, and carbon-to-nitrogen ratio 9:1. Under these conditions, sporulation increased twelve times compared with control and this result was in agreement with model predictions.
Subject(s)
Ophiostomatales/growth & development , Spores, Fungal/growth & development , Carbon/metabolism , Culture Media/chemistry , Dose-Response Relationship, Drug , Hydrogen Peroxide/metabolism , Nitrogen/metabolism , Ophiostomatales/drug effects , Spores, Fungal/drug effects , Time FactorsABSTRACT
The viability of conidia of Esteya vermicola, a potentially important biocontrol agent against the pinewood nematode Bursaphelenchus xylophilus, is usually determined by cultivation for 18-48 h in culture medium. As an alternative to this labor-intensive method, we have developed a rapid, simple, and low-cost staining method for assessing E vermicola conidia survival rates. A mixture of neutral red and methylene blue was found to be the most optimal among several stains that also included safranin O and Janus green B. This mixture stained nonviable conidia blue, in contrast to viable conidia, which were stained red in the cytoplasm and blue in the cell wall. This method may be particularly useful for traditional research laboratories, as it provides rapid results using common, relatively inexpensive laboratory equipment.
Subject(s)
Ophiostomatales/physiology , Pinus/parasitology , Plant Diseases/prevention & control , Spores, Fungal/physiology , Tylenchida/microbiology , Animals , Azo Compounds , Biological Control Agents , Coloring Agents , Phenazines , Staining and LabelingABSTRACT
Ginsenosides, the secondary plant metabolites produced by Panax ginseng are responsible for the enhancing effects on learning observed following treatment with Panax ginseng. A number of studies have provided correlational evidence that cell proliferation and survival are closely associated with hippocampal-dependent learning tasks. In this study, to investigate the beneficial effects of ginsenoside Rh1 on hippocampal cells and learning, mice (6 months old) were administered ginsenoside Rh1 at a dose of 5 and 10 mg/kg/day for a period of 3 months. Saline-treated mice were used as controls. The enhancement of memory and learning in the mice was evaluated by hippocampal-dependent tasks (passive avoidance tests and Morris water maze tests) and the immunohistochemical marker of cell proliferation, bromodeoxyuridine (BrdU). In addition, the levels of brain-derived neurotrophic factor (BDNF) were measured following treatment. Based on our data, the Rh1-treated group (5 and 10 mg/kg) showed a significantly improved learning and memory ability in the passive avoidance tests compared with the control group; however, only treatment with 10 mg/kg ginsenoside Rh1 significantly promoted spatial learning ability in the Morris water maze test. Ginsenoside Rh1 significantly enhanced cell survival in the dentate gyrus of mice, although it did not enhance hippocampal cell proliferation. In addition, ginsenoside Rh1 upregulated the expression of BDNF. These findings address the potential therapeutic significance of ginsenoside Rh1 as a nutritional supplement in memory loss and neurodegenerative diseases.
Subject(s)
Dentate Gyrus/drug effects , Ginsenosides/pharmacology , Maze Learning/drug effects , Memory/drug effects , Animals , Brain-Derived Neurotrophic Factor/genetics , Brain-Derived Neurotrophic Factor/metabolism , Cell Proliferation/drug effects , Cell Survival/drug effects , Dentate Gyrus/cytology , Dentate Gyrus/metabolism , Ginsenosides/administration & dosage , Male , Mice , Mice, Inbred ICR , Panax/chemistry , Up-RegulationABSTRACT
A wide range of plant foods and dietary supplements are able to modify the functioning of the central nervous system. In the present study, we observed that oral administration of ginsenoside Rh2 (10 mg/mL) for 3 weeks significantly improved spatial learning and memory. Spatial memory and learning was evaluated in mice by hippocampus-dependent tasks (Morris water maze test) and immunohistochemical marker of cell genesis bromodeoxyuridine. Ginsenoside Rh2 treatment (30 days) promoted cell survival and genesis. Further, ginsenoside Rh2 treatment in enriched condition had no significant effects on cell survival compared with standard condition exposure. These results revealed that ginsenoside Rh2-mediated spatial learning and memory improvement was associated with cell genesis and survival and may be parallel to the mechanism of environmental enrichment. Therefore, ginsenoside Rh2 may have efficacy as a dietary supplement for spatial learning and memory improvement.
Subject(s)
Ginsenosides/administration & dosage , Learning/drug effects , Memory/drug effects , Plant Extracts/administration & dosage , Animals , Hippocampus/drug effects , Hippocampus/physiology , Male , Mice , Mice, Inbred ICRABSTRACT
Esteya vermicola, an endoparasitic fungus of Bursaphelenchus xylophilus, the pinewood nematode (PWN), exhibits great potential as a biological control agent against this nematode. E. vermicola produces blastospores in liquid media and aerial conidia on solid media. The agent was mass-produced using two kinds of culture media: S (50 % wheat bran and 50 % pine wood powder), L (0.5 g wheat bran and 0.5 g pinewood powder in 200 ml of potato dextrose broth), and two controls: SC (potato dextrose agar), LC (potato dextrose broth). Yields, multiple stress tolerance, storage life, new generation conidial number, and PWN mortality rates of the spores were measured in each of these four media and compared. The spore yields, new generation conidial number, and nematode mortality rates of blastospores were higher than those of conidia. Nevertheless, the conidia had a higher germination rate than the blastospores during the storage process and multiple stress treatments. Considering the number of spores surviving from the process of the storage and multiple stress treatments per unit of mass media, the blastospores from L survived most. Comprehensive analysis indicates that the L culture medium is the most optimal medium for mass production relatively.
Subject(s)
Ascomycota/physiology , Spores, Fungal/physiology , Tylenchida/microbiology , Animals , Culture Media , Fermentation , Pinus/parasitology , Plant Diseases/parasitology , Stress, Physiological , Tylenchida/growth & development , Wood/parasitologyABSTRACT
Chemotherapy frequently results in neurocognitive deficits that include impaired learning and memory. Thus, it is important to prevent or ameliorate the persistence of cognitive impairment. Compound K was employed to examine the ameliorating effect on chronic treatment with cyclophosphamide. Eight week-old ICR mice were given 80 mg/kg cyclophosphamide, cyclophosphamide combined with compound K (2.5, 5 and 10 mg/kg) or saline injections once per week for 4 weeks. Passive avoidance test and Y maze were used to evaluate memory and learning ability. Immunohistochemical staining for progenitor cell and immature neurons was used to assess changes in neurogenesis. Compound K (10 mg/kg) is able to ameliorate the decrease of neurogenesis in the hippocampus caused by cyclophosphamide. These results suggest that compound K might be a potential strategy to ameliorate or repair the disrupted hippocampal neurogenesis induced by the side effect of chemotherapy agent.
Subject(s)
Antineoplastic Agents/adverse effects , Cognition Disorders/drug therapy , Cyclophosphamide/adverse effects , Ginsenosides/pharmacology , Animals , Antineoplastic Agents/pharmacology , Cognition Disorders/chemically induced , Cyclophosphamide/pharmacology , Disease Models, Animal , Hippocampus/drug effects , Hippocampus/metabolism , Male , Maze Learning , Memory/drug effects , Mice , Mice, Inbred ICR , Neurons/drug effects , Neurons/metabolism , Stem Cells/cytology , Time FactorsABSTRACT
The nematophagous fungus Esteya vermicola has tremendous potential for biological control. This species exhibits strong infectious activity against pinewood nematodes, whereas the study on the effect of nutrition and environmental factors is still of paucity. Carbon (C), nitrogen (N), pH value, temperature, and water activity have great impact on the fungal growth, sporulation, and germination. In nutrition study, the greatest number of conidia (2.36 × 10(9) per colony) was obtained at the C:N ratio of 100:1 with a carbon concentration 32 g l(-1). In addition, the germination rate and radial growth of E. vermicola were used to evaluate the effects of environmental conditions and they were optimized as following: pH 5.5, 26 °C and water activity of 0.98. Our results also confirmed that variation of environmental factors has a detrimental influence on the efficacy of active conidia and growth of fungus. Moreover, under above optimal condition, the biocontrol efficacy was significantly improved in regard to the increase of adhesive and mortality rate, which highlight the study on the application of E. vermicola as pine wilt disease biocontrol agent.
Subject(s)
Ophiostomatales/growth & development , Ophiostomatales/metabolism , Carbon/metabolism , Hydrogen-Ion Concentration , Nitrogen/metabolism , Pinus/microbiology , Spores, Fungal/growth & development , Temperature , Water/metabolismABSTRACT
This paper studied the variations of leaf photosynthesis gas exchange, chlorophyll fluorescence, and free amino acid contents of Heyedysarum scoparium under the stress of different concentration NaCl, aimed to understand the effects of salt stress on the leaf physiological characteristics and free amino acid metabolism of the plant. Under mild salt stress, the photo-damage of the leaf photosystem II was avoided via the dissipation of excess excitation energy, and stomatal limitation was the main factor reducing the photosynthesis rate. With the increase of salt stress, the leaf photoprotection was not sufficient to avoid oxidative damage, and thus, the damage to photosystem II happened. Under the stress of 200 mmol NaCl x L(-1), non-stomatal limitation was the main factor responsible for the inhibition of photosynthesis. The accumulation and metabolism of major free amino acids in H. scoparium leaves varied under salt stress. Under mild salt stress, the major free amino acids such as proline, glutamate, aspartate, and alanine had a significant accumulation, but with the further increase of salt stress, the aspartate and alanine contents in H. scoparium leaves decreased, while the synthesis and accumulation of praline increased.
Subject(s)
Amino Acids/metabolism , Fabaceae/physiology , Photosynthesis/physiology , Sodium Chloride/pharmacology , Stress, Physiological/physiology , Fabaceae/metabolism , Plant Leaves/physiologyABSTRACT
In the present investigation, we successfully employed a cell-free extract of Esteya vermicola CNU 120806 to convert ginsenoside Rg3 to Rh2. Three important factors including pH, temperature and substrate concentration were optimized for the preparation of Rh(2). The optimal condition was obtained as follows: 50°C, pH 5.0 and substrate concentration of 3 mg ml(-1). The yield of conversion was up to 90.7%. In order to identify the specificity of the ß-glucosidase activity of Esteya vermicola CNU 120806, ginsenoside Re (protopanaxatriol saponins) was treated under the same reaction system. Interestingly, no new metabolite was generated, which elucidated that the enzymatic process only occurred by hydrolysis of the terminal glucopyranosyl moieties at the C-3 carbon of ginsenoside Rg(3). The crude enzyme extract can be used for commercial ginsenoside Rh(2) preparation.