ABSTRACT
Tremella fuciformis Berk (TF) is a common edible and medicinal mushroom, and has long been used in food and in Chinese medicine. It possesses anticancer, anti-inflammation, anti-oxidative, and neuroprotective abilities. Since their cultivation is a problem, TFs in Taiwan are primarily imported from China, which has a problem with pesticide residues. Thus, the question of whether the Taiwan cultivated TFs, T1, and T6 showed similar or even better results than TFs from China (CH) was assessed in the present study. The results of the physicochemical tests of these TFs showed that T1 extracted by hot water (T1H) has the highest concentration of polysaccharide; meanwhile, T6 extracted by cold water (T6C) showed the highest amount of protein. Regarding the immune modulatory effects of these TFs, hot water extracts of these TFs augmented significantly the inducible nitric oxide synthetase (iNOS), interleukin (IL)-6, and tumor necrosis factor (TNF)-[Formula: see text] mRNA expression than those of cold water extracts. On the other hand, the cold water extracts of TFs, especially of T1C, obviously suppressed cancer cell survival better than those of hot water extracts. Interestingly, we found that hot water extracts of TFs may augment necrotic cell death, whereas, cold water extracts of TFs induce apoptosis. Furthermore, we also showed that these TFs activate caspase-3 cleavage, up regulate the Bax/Bcl-2 ratio, and decrease MMP-9 expressions in PC-3 cells. Taken together, our results indicated that T1 and T6 strains of TFs showed the similar immune modulatory and anticancer abilities were better than the CH strain of TFs.
Subject(s)
Agaricales/chemistry , Antineoplastic Agents, Phytogenic , Immunologic Factors , Plant Extracts/chemistry , Plant Extracts/pharmacology , Prostatic Neoplasms/pathology , Animals , Apoptosis/drug effects , Caspase 3/metabolism , Cell Death/drug effects , Cell Line, Tumor , Fungal Polysaccharides/isolation & purification , Gene Expression/drug effects , Humans , Interleukin-6/genetics , Interleukin-6/metabolism , Male , Matrix Metalloproteinase 9/metabolism , Mice , Necrosis , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , Plant Extracts/isolation & purification , Prostatic Neoplasms/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , RAW 264.7 Cells , RNA, Messenger/metabolism , Temperature , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Up-Regulation/drug effects , Water , bcl-2-Associated X Protein/metabolismABSTRACT
OBJECTIVE: To study the protective effect of propofol precondition against glutamate (Glu) neurotoxicity to neonatal rat cerebrocortical slices. METHODS: Brain slices of Sprague-Dawley (SD) rats were cultured in vitro and observed the morphologic changes. Brain slices were randomly divided into three groups: blank control group, Glu injury group (1 mmol/L Glu for 0.5 hour), propofol precondition group (20 mg/L propofol for 24 hours), each n=12. Changes in pathological and ultra-structure of cells were observed using microscope. Lactate dehydrogenase (LDH) leakage rate was measured. Meanwhile, the expression of glial fibrillary acidic protein (GFAP) was detected by immunohistochemical technology, then the positive cell were counted. RESULTS: Cultured brain slices of cell were intact and survived well. Hematoxylin-eosin (HE) staining, electron microscopy and LDH test results showed that cerebral film neuron severely damage, gliosis, edema, LDH leakage rate in Glu injury group were significantly more severe compared with blank control group [(68.5±2.0)% vs. (16.0±2.5)%, P<0.01]. Reduce the brain slice of the propofol pretreatment group of neuronal cell jury, cell shape recovery significantly reduced LDH leakage rate compared with the Glu injury group [(38.5±2.4)% vs. (68.5±2.0)%, P<0.05]. Immunohistochemical detection of GFAP expression of Glu injury group glial cell body swelling, producing increase in the number of GFAP positive reaction strong, the number of positive cells compared with blank control group was significantly increased (50±5 cells/HP vs. 10±3 cells/HP, P<0.01). The recovery of propofol pretreatment group glial cell morphology, cell processes slender GFAP positive reaction decreased the number of positive cells compared with the Glu injury group was significantly decreased (30±4 cells/HP vs. 50±5 cells/HP, P<0.05). CONCLUSION: Propofol pretreatment has protective effect against Glu injured rat cerebrocortical slices.
Subject(s)
Brain/drug effects , Glutamic Acid/adverse effects , Propofol/pharmacology , Animals , Brain/metabolism , Brain/pathology , L-Lactate Dehydrogenase/metabolism , Neurons/drug effects , Neurons/metabolism , Rats , Rats, Sprague-Dawley , Tissue Culture TechniquesABSTRACT
Signaling pathways, specifically calcium and calcium-dependent protein kinase (CDPK), have been implicated in the regulation of stress and developmental signals in plants. Here, we reported the isolation and characterization of an orchid, Phalaenopsis amabilis, CDPK gene, PaCDPK1, by using the rapid amplification of cDNA ends (RACE)-PCR technique. The full length cDNA of 2,310 bp contained an open reading frame for PaCDPK1 consisting of 593 amino acid residues. Sequence alignment indicated that PaCDPK1 shared similarities with other plant CDPKs. PaCDPK1 transcripts were expressed strongly in labellum but not in leaves and roots. In addition, the PaCDPK1 gene was transcriptionally activated in response to low temperature, wounding, and pathogen infection. To identify the regulatory role of the PaCDPK1 promoter, a construct containing the PaCDPK1 promoter fused to a beta-glucuronidase (GUS) gene was transferred into Arabidopsis by Agrobacterium-mediated transformation. GUS staining revealed that PaCDPK1/GUS expression was induced by cold, wounding, and pathogen challenge in leaves and stems of transgenic Arabidopsis. These results suggested that this PaCDPK1 gene promoter could be used as an endogenous promoter for biotechnological purposes in orchids.
