Objective : The water extract of Danguiyonghoihwan (DGYHW) has been traditionally used in treatment of tinnitus in Oriental Medicine. However, little is known about the mechanism by which DGYHW rescues auditory neuronal cells from injury damages. Therefore, in this study I effort to elucidate the mechanism of the cytoprotective effect of the DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Methods : I determined the elevated cell viability by DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Glutamate induced neuronal damage in oranotypic explant culture also, glutamate decreased cell viability on VOT-33 cells but pretreatment with DGYHW inhibited cell death. Results : One of the main mediator of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pretreatment with DGYHW inhibited this ROS generation from glutamated-stimulated VOT-33 cells. Also, I identified that the ROS-induced DCF-DA green fluorescence is reduced by DGYHW pretreatment. The critical markers of apoptotic cell death were cleavages of procaspase-3 protease protein. So I checked the expression level and cleavage of procaspase-3 protease protein. Glutamate-treated VOT-33 cells were shown to have cleavage of procaspase-3 protease proteins and following reduction of expression of these proteins. But I found that pre-treatment with DGYHW protects glutamate-induced changes of biochemical marker protein, caspase-3. Conclusion : These findings indicated that DGYHW may prevent cell death from glutamate induced VOT-33 cell death by inhibiting the ROS generation and modulation of protein expressions in procaspase-3, catalase and Bcl-2.
Objectives Ethanol is a potent inhibitor of muscle protein synthesis. Muscle mass is regulated by the balance between rates of protein synthesis and protein breakdown. Both acute and chronic alcohol consumption inhibits synthesis to a greater extent than degradation. Protein synthesis is more intensely decreased in type II fibers than in type I fibers. Apoptosis has been shown to occur frequently in a variety of tissues in response to chronic alcohol feeding. Increased muscle fiber apoptosis has been shown in alcoholics with myopathy. Pueraria radix has been used for many disorders such as fevers, gastrointestinal disorders, muscle aches, allergies, respiratory problems, skin problems, high blood pressure, migraine headaches, lowering cholesterol and treating chronic alcoholism. We therefore tested the hypothesis that oral treatment with Pueraria radix could reduce the ethanol-induced muscle atrophy. Methods Young male Sprague-Dawley rats were orally given 25% ethanol (5 ml/kg, body weight) daily with Ethanol for 4 weeks. Normal group was similarly administrated with saline. The Rats of Pueraria radix treated group (EtOH+PR) were orally administrated Pueraria radix water extract, and rats of EtOH group were given with the vehicle only. After 4 week, the morphology of gastrocnemius and plantaris muscles were assessed by hematoxylin and eosin staining. The immunoreactivities of pre-apoptotic BAX and anti-apoptotic Bcl-2 proteins were also measured. Results The muscles from rats of EtOH group represented a significant reduction in average cross section area compared to Normal group. EtOH+PR group had increased fiber compared to the EtOH group. Moreover, to investigate the ethanol-induced muscular apoptosis, the immunohistochemical analysis of Bax and Bcl-2 was carried out. The treatment with Pueraria radix (EtOH+PR) significantly decreased BAX expression and increased Bcl-2 expression 4 weeks after ethanol administration when compared with Normal group. Conclusions These results suggest that Pueraria radix water extract has protective effects on chronic alcohol induced myopathy.
Effects of sodium bicarbonate (SBC) injection on meat quality and functionality of porcine M. longissimus lumborum were investigated. Fifteen pigs ($100{\pm}5kg$) were randomly selected at a commercial slaughter plant. After slaughtering the loins were dissected from the carcass before chilling at approximately 30 minutes post mortem. The loins were divided into four cuts for sample treatment, and SBC of 0.25 M, 0.40 M and 0.75 M was injected (2% w/w) using a syringe. As SBC injection level was increased, muscle pH increased significantly (p<0.05). SBC injection decreased lightness ($L^*$) values on the surface of muscle. Moreover, with injection of SBC, drip loss %, cooking loss % and shear force were significantly (p<0.05) decreased, whereas WHC and $Na^+$ content were significantly (p<0.05) increased. From panel testing of uncooked pork loin, no significant differences (p>0.05) were found in aroma, off-flavor and drip between injection of SBC at different levels and the control, although color and acceptability were significantly lower (p<0.05) in control pork loin compare with injection of SBC at 0.75 M. In cooked pork loin from the panel test, aroma, flavour, off-flavour and juiciness were found to be similar (p>0.05) on all treatments, but tenderness and acceptability were significantly higher (p<0.05) with injection of SBC at 0.75 M than for control loin. Myofibrillar protein solubility of muscles treated with SBC was significantly (p<0.05) higher than that of the control, although no significant differences (p>0.05) were found in sarcoplasmic protein solubility between the treatments. These results indicated that SBC injection into pre-rigor porcine M. longissimus lumborum could improve ultimate pork quality characteristics such as meat color, water-holding capacity, and could inhibit muscle protein denaturation due to an increase in muscle pH.
The aim of this study was to examine the effectiveness of pre-cooking conditions on the quality characteristics of ready-to-eat (RTE) Samgyetang. Raw chickens were steamed under the different conditions of 50℃/30 min (T1), 65℃/30 min (T2), 85℃/30 min (T3), and 90℃/10 min (T4) prior to retorting at 120℃ for 65 min. The results showed that pre-cooking conditions in all treated samples could reduce fat contents in breast and leg meats by 8.5-11.7% and 10.0-11.0% compared to the control, even though there were no significant differences among treatments (p>0.05). The L* and b* values of breast and leg meats treated with the higher temperature and longer time conditions were significantly higher than the control (p<0.05), while a* values tended to decrease despite of not to a significant extent (p>0.05). Moreover, apparent viscosity and water soluble protein showed insignificant differences (p>0.05) among the samples as a result of the retorting process, which might have more negative influences on the quality. T2 samples obtained significantly the highest average Quantitative Descriptive Analysis (QDA) score and transmittance value, representing the most clear broth among the samples, compared to the control. On the other hand, T3 showed the highest cooking loss among the treatments and the lowest QDA scores among the samples. In conclusion, pre-cooking treatment prior to retorting in manufacturing Samgyetang is a plausible way to reduce its fat content. A pre-cooking condition at either 65℃ for 30 min, or 90℃ for 10 min are recommended for producing Samgyetang with optimum quality.
Objectives : Ganoderma lucidum(Ganoderma or lingzhi, 靈芝) is a well-known oriental medical mushroom containing many bioactive compounds. The possible mechanisms involved in its effects on cancer cells remain to be elucidated. In the present study, the anti-proliferative and apoptotic activities of the G. lucidum ethanol extract(GEE), in AGS human gastric cancer cells were investigated. Methods : It was found that exposure of AGS cells to GEE resulted in the growth inhibition in a dose and time dependent manner as measured by trypan blue count and MTT assay. The anti-proliferative effect of GEE treatment in AGS cells was associated with morphological changes and formation of apoptotic bodies, and the flow cytometry analysis confirmed that GEE treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptosis of AGS cells by GEE were connected with a concentration and time-dependent up-regulation of tumour necrosis factor-related apoptosis-inducing ligand(TRAIL) expression. Results : The levels of XIAP and survivin expression, members of IAP family proteins, were gradually down-regulated by GEE treatment. However other members of IAP family proteins such as cIAP-1 and cIAP-2 remained unchanged in GEE-treated AGS cells. GEE treatment also induced the proteolytic activation of caspase-3, caspase-8 and caspase-9 and a concomitant degradation of poly(ADP-ribose) polymerase(PARP) protein, a caspase-3 substrate protein. Additionally, GEE-induced apoptosis was associated with the inhibition of Akt activation in a concentration and time-dependent manner, and pre-treatment with LY294002, a phosphoinositide 3-kinase(PI3K)/Akt inhibitor, significantly increased GEE-induced growth inhibition and apoptosis. Conclusions : Therefore, G. lucidum has a strong potential as a therapeutic agent for preventing cancers such as gastric cancer cells.
The pistil of nelumbo nucifera (PNN) is used in the treatment of nocturnal pollution, hematemesis, epistaxis, metrorrhagia and diarrhoea in traditional medicine. The present study was examined to evaluate the effects of PNN on the production of pro-inflammatory mediators in vitro. After the treatment of PNN, cell viability was measured by MTT assay, nitric oxide (NO) production was monitored by measuring the nitrite content in culture medium. The protein bands were determined by immunoblot analysis and levels of cytokines were analyzed by sandwich immunoassays. In the MTT assay, the doses of PNN extract (0.03, 0.10 mg/ml) had no significant cytotoxicity. The increases of NO production and inducible nitric oxide synthase expression were detected in lipopolysaccharide(LPS)-activated Raw 264.7 cells compared with control, in contrast, these increases were significantly attenuated by pre-treatment with PNN. In cytokine assay, the massive pro-inflammatory cytokines such as tumour necrosis factor-$\alpha$, interleukin (IL)-$1{\beta}$ and IL-6 were induced in LPS-activated Raw 264.7 cells, but pre-treatment of Raw 264.7 cells with PNN caused inhibition (TNF-$\alpha$=14.17%, IL-$1{\beta}$=107.43%, IL-6=46.27%) the production of cytokines by LPS. In addition, PNN reduced prostaglandin E2 productions in a dose-dependent manner (0.03mg/ml=37.52%, 0.10 mg/ml=83.77%) as a consequence of the inhibition of cyclooxygenase-2 expression. Taken together, our data indicates that PNN can regulate the inflammatory response in macrophage cells activated by Gram-negative infection.
Chondroitin sulfate proteoglycan (CSPG) inhibits neurite outgrowth of various neuronal cell types, and CSPG-associated inhibition of neurite outgrowth is mediated by the Rho/ROCK pathway. Mesenchymal stromal/stem cells (MSCs) have the potential to differentiate into neuron-like cells under specific conditions and have been shown to differentiate into neuron-like cells by co-treatment with the ROCK inhibitor Y27632 and the hypoxia condition mimicking agent $CoCl_2$. In this study, we addressed the hypothesis that a ROCK inhibitor might be beneficial to regenerate neurons during stem cell therapy by preventing transplanted MSCs from inhibition by CSPG in damaged tissues. Indeed, dose-dependent inhibition by CSPG pretreatment was observed during morphological changes of Wharton's jelly-derived MSCs (WJ-MSCs) induced by Y27632 alone. The formation of neurite-like structures was significantly inhibited when WJ-MSCs were pre-treated with CSPG before induction under Y27632 plus $CoCl_2$ conditions, and pretreatment with a protein kinase C inhibitor reversed such inhibition. However, CSPG treatment resulted in no significant inhibition of the WJ-MSC morphological changes into neuron-like cells after initiating induction by Y27632 plus $CoCl_2$. No marked changes were detected in expression levels of neuronal markers induced by Y27632 plus $CoCl_2$ upon CSPG treatment. CSPG also blocked the morphological changes of human bone marrow-derived MSCs into neuron-like cells under other neuronal induction condition without the ROCK inhibitor, and Y27632 pre-treatment blocked the inhibitory effect of CSPG. These results suggest that a ROCK inhibitor can be efficiently used in stem cell therapy for neuronal induction by avoiding hindrance from CSPG.
Kang, Seong Hee;Bak, Dong-Ho;Chung, Byung Yeoup;Bai, Hyoung-Woo;Kang, Bo Sun
The Korean Journal of Physiology and Pharmacology
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제24권5호
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pp.413-422
/
2020
Delphinidin is a major anthocyanidin compound found in various vegetables and fruits. It has anti-oxidant, anti-inflammatory, and various other biological activities. In this study we demonstrated the anti-cancer activity of delphinidin, which was related to autophagy, in radiation-exposed non-small cell lung cancer (NSCLC). Radiosensitising effects were assessed in vitro by treating cells with a subcytotoxic dose of delphinidin (5 μM) before exposure to γ-ionising radiation (IR). We found that treatment with delphinidin or IR induced NSCLC cell death in vitro; however the combination of delphinidin pre-treatment and IR was more effective than either agent alone, yielding a radiation enhancement ratio of 1.54 at the 50% lethal dose. Moreover, combined treatment with delphinidin and IR, enhanced apoptotic cell death, suppressed the mTOR pathway, and activated the JNK/MAPK pathway. Delphinidin inhibited the phosphorylation of PI3K, AKT, and mTOR, and increased the expression of autophagy-induced cell death associated-protein in radiation-exposed NSCLC cells. In addition, JNK phosphorylation was upregulated by delphinidin pre-treatment in radiation-exposed NSCLC cells. Collectively, these results show that delphinidin acts as a radiation-sensitizing agent through autophagy induction and JNK/MAPK pathway activation, thus enhancing apoptotic cell death in NSCLC cells.
Sunagawa, K.;Hashimoto, T.;Izuno, M.;Hashizume, N.;Okano, M.;Nagamine, I.;Hirata, T.
Asian-Australasian Journal of Animal Sciences
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제21권4호
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pp.538-546
/
2008
Large-type goats eating dry forage secreted large volumes of saliva which resulted in the loss of $NaHCO_3$ from the blood and decreased plasma volume (hypovolemia). This research investigated whether or not the loss of $NaHCO_3$ from the blood and hypovolemia brought about by dry forage feeding actually depresses feed intake in large-type goats under free drinking conditions. The present experiment consisted of three treatments (NI, ASI, MI). All treatments in this experiment were carried out under free drinking conditions. In the NI control (NI), a solution was not infused. In the ASI treatment, i.v. infusion of artificial saliva was initiated 2 h before feeding and was continued for a total of 3 h concluding 1 h after the commencement of the feeding perod. In the MI treatment, mannitol solution was infused to replenish only water lost from the blood in the form of saliva. The hematocrit and plasma total protein concentrations during feeding in the NI control were observed to be higher than pre-feeding levels. This indicated that dry forage feeding-induced hypovolemia was caused by the accelerated secretion of saliva during the initial stages of feeding in freely drinking large-type goats. Increases in hematocrit and plasma total protein concentrations due to dry forage feeding were significantly suppressed by the ASI treatment. While hematocrit during feeding in the MI treatment was significantly lower than the NI control, plasma total protein concentrations were not different. From these results, it is clear that the MI treatment was less effective than the ASI treatment in mitigating the decreases in plasma volume brought about by dry forage feeding. This indicates that plasma volume increased during dry forage feeding in the ASI treatment which inhibited production of angiotensin II in the blood. The ASI treatment lessened the levels of suppression on dry forage feeding, but the MI treatment had no effect on it under free drinking conditions. The results indicate that despite the free drinking conditions, increases in saliva secretion during the initial stages of dry forage feeding in large-type goats caused $NaHCO_3$ to be lost from the blood into the rumen which in turn caused a decrease in circulating plasma volume and resulted in activation of the renin-angiotensin system and thus feeding was suppressed.
Objective: The effectiveness of Staphylococcus protein A (SPA) in improving the penetration ability of sperm and reducing antisperm antibody (ASA) titers in immunologically infertile males was evaluated. Methods: Seminal fluid samples were obtained from 15 infertile men, and ASA titers were assessed with the latex agglutination test. Identification of immunoglobulin (Ig) classes and characterization of the antigens involved in the immune response were performed using indirect immunofluorescence. Local ASAs typically present as a mixture of IgG and IgA classes. The capillary tube penetration method was used to assess the capability of spermatozoa to penetrate the cervical mucus (CM). Results: ASAs associated with the neck region of sperm showed a significantly lower migration distance in the CM of infertile females than ASAs associated with the head or tail segments. ASA-positive seminal fluid exhibited significant increases in the mean migration distance (2.6 ± 1.4 cm vs. 1.54 ± 1.1 cm, respectively; p< 0.001) and sperm concentration (174 ± 121.0 × 103/mL vs. 101 ± 93.7 × 103/mL, respectively; p= 0.033) after treatment with SPA compared to pre-treated samples. A significant reduction (p< 0.01) in the recorded ASA titer was detected. Conclusion: These results indicate that SPA can be used as a sorting regimen for insemination programs. However, further studies are warranted to assess its influence on pregnancy rate.
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