• Journal of Veterinary Clinics
• /
• v.11 no.2
• /
• pp.545-552
• /
• 1994

This experiment was carried out to investigate the renditions to maintain good post-thaw motility and viability of canine spermatozoa when the semen was frozen using methanol. The semen from two male dogs which had been proven to be fertile in the previous one year was treated with different compositions of semen diluent and was frozen at different freezing temperatures, When canine semen was frozen at-2$0^{\circ}C$, -6$0^{\circ}C$, or -8$0^{\circ}C$, the spermatozoa frozen and stored at -2$0^{\circ}C$ showed very low post-thaw motility and viability from day 2 to 7 and showed no viability since day 15 after freezing. The spermatozoa frozen and stored at -6$0^{\circ}C$ or -8$0^{\circ}C$ showed higher post-thaw motility and viability on day 2, 1, 15 and 30 after freezing than that frozen and stored at-2$0^{\circ}C$(p<0.01), with no difference between two groups. Among different composition groups of the semen diluents of control(tris + egg yolk + glycerol), egg yolk-free, 히ycerol-free, and tris-free, Prior to freezing, the egg yolk-free diluent showed significantly love. motility and viability than the other diluents(p<0.05). On each thawing day (from day 2 to 15 after freezing), control group showed considerably higher motility and viability than the other groups(p<0.01). The canine spermatozoa frozen and stored at -6$0^{\circ}C$ and -8$0^{\circ}C$ showed gradual decrease of motility from day 2 to 30 after freezing and the spermatozoa of these two groups thawed on day 30 showed considerably love. motility than those thawed on day 2 after freezing, respectively(p<0.01). These results indicate that the freezing temperature of either -6$0^{\circ}C$ or -8$0^{\circ}C$ can be applicable to the freezing method using methanol and also all of the components of the semen diluent including cryoprotectant, buffer and cold-shock buffer are very important to maintain motility and viability of canine spermatozoa in the freezing and thawing procedure.

• Journal of Embryo Transfer
• /
• v.28 no.4
• /
• pp.373-379
• /
• 2013

Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or $37^{\circ}C$. In case of ICR strain, primary TSCs cultured at $37^{\circ}C$ showed significantly higher proliferation and viability than those at $35^{\circ}C$ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the $35^{\circ}C$ culture condition. Moreover, sub-passage of primary TSCs at $35^{\circ}C$ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at $35^{\circ}C$, which showed no significant difference in the viability, compared to those at $37^{\circ}C$. Furthermore, sub-passaged TSCs cultured at $37^{\circ}C$ showed no significant differences in proliferation and viability, compared to those at $35^{\circ}C$. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at $35^{\circ}C$, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at $37^{\circ}C$. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.

• The Journal of Korean Medicine
• /
• v.21 no.2
• /
• pp.79-86
• /
• 2000

Objectives and Methods : In order to elucidate toxic mechanism of myocardial damage and protective effect of myrrha water extract against cytotoxic effect of xanthine oxidase/hypoxanthine(XO/HX), cardioprotective effect of myrrha water extract was examined by MTT assay, LDH (Lactate Dehydrogenase) activity and heart beating rate after cultured myocardial cells derived from neonatal mouse were treated with various concentration of XO/HX, a free radical. Results : XO/HX induced a decrease of cell viability, an increase in the amount of LDH, and a decrease of heart beating rate on cultured myocardial cells in a dose-dependent manner. In cardioprotective effect of myrrha water extract, it showed a decrease in the amount of LDH and an increase of heart beating rate on cultured myocardial cells damaged by XO/HX. Conclusions : From the above results, it is suggested that XO/HX showed toxic effect in cultured myocardial cells derived from neonatal mouse and that myrrha water extract is very effective in the prevention of XO/HX-induced cardiotoxicity.

• International Journal of Oral Biology
• /
• v.35 no.3
• /
• pp.99-106
• /
• 2010

Luteolin is a flavonoid that exists in a glycosylated form in celery and green pepper. Flavonoids possess antioxidant and anti-inflammatory properties and can reduce the expression of key inflammatory molecules in macrophages and monocytes. It has been reported also that some flavonoids have effects on bone metabolism. The effects of luteolin on the function of osteoblasts were investigated by measuring cell viability, alkaline phosphatase activity, type I collagen production, osteoprotegerin secretion, Wnt promoter activity, BMP-2 and Runx2 expression and calcified nodule formation. Luteolin has no effects upon osteoblast viability but induced an increase in alkaline phosphatase activity, type I collagen production and a decrease in osteoprotegerin secretion in these cells. Luteolin treatment also upregulated BMP-2 mRNA expression. These results suggest that luteolin may be a regulatory molecule that facilitates the differentiation of osteoblasts.

• Toxicological Research
• /
• v.11 no.1
• /
• pp.103-108
• /
• 1995

Rat renal proximal tubule suspension was prepared from adult male Sprague Dawley rat (250-300g) by mechanical (non-enzymatical) method and evaluated as a pontential model for mechanistic studies and early screening of nephrotoxicity, using anionic antibiotics (cephaloridine). Cephaloridine (CPL) produced an increase in LDH release into media. This release results from decrease a proximal tubule cell viability and subsequently increase the permeability of cell viability and subsequently increase the permeability of cell membrane. Since loss of intracellular potassium and ATP into media is the sign of disruption of cell membrane, especially basolateral membrane (BLM), CPL induced proximal tubule cell compromise also appear be associated with BLM, maybe $Na^+-K^+$ ATPase. Also seen was significant depression in brush border membrane (BBM) ALP activity and no significantly increase in BBM GGT activities. The inhibition of typical anion, PAH accumulation (especially, CPL 5 mM) and cation, TEA (especially, 4hours incubation) were seen dose dependently. This is because of CPL accumulation in renal proximal tubule and increase of cytotoxicity.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.22 no.2
• /
• pp.50-59
• /
• 2009

Objectives : The aim of this study was conducted that CRE (Coptidis Rhizoma Extract) induces apoptosis in YD-10B cells, human oral squamous carcinoma cell line. Methods : In this study, YD-10B cells were exposed to CRE (0.03-0.30 mg/ml), for 6-24 hours. We measured the effects of CRE on the changes of cell viability and cell membrane, TUNEL assay of CRE-treated YD-10B cell. Results : In this study, CRE caused a decrease of viability in YD-10B cells, human oral squamous carcinoma cell line. When YD-10B cells were treated with CRE, cells showed dose-dependent manner apoptotic cell death. Conclusions : These results suggest that CRE may be potential therapeutic approach in the clinical management of oral squamous cell carcinoma.

• Biomedical Science Letters
• /
• v.19 no.2
• /
• pp.83-89
• /
• 2013

Parkin is a protein known to have tumor suppressive functions. In a previous study, we determined that Parkin expression restores susceptibility to TNF-${\alpha}$-induced death in HeLa cells. ${\beta}$-catenin is a key protein in the Wnt signaling pathway and excessive activation of the ${\beta}$-catenin pathway can promote cancer development. In this study, we found that ${\beta}$-catenin levels decreased dramatically in Parkin over-expressing HeLa cells treated with TNF-${\alpha}$. We used chemical inhibitors of cell signaling pathways to identify the signaling molecules involved in ${\beta}$-catenin down-regulation. Our results indicate that the PKC inhibitor (RO-31-7549) blocked parkin-induced down-regulation of ${\beta}$-catenin. We also show that Parkin-induced decrease in cell viability in TNF-${\alpha}$-treated HeLa cells is alleviated upon treatment with a PKC inhibitor. Taken together, these results suggest the possibility that ${\beta}$-catenin reduction may be associated with Parkin-induced decrease of cell viability in TNF-${\alpha}$ treated HeLa cells.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.16 no.2
• /
• pp.289-295
• /
• 2002

To certify the protective effect of herbal medicine against oxygen free radical-induced myocardiotoxicity, cytotoxicity was measured using MTT, LDH activity and Beating rate assay in the presence of Guaruhaebaekbanha-tang(GHBT) extracts or single constituents of this prescription. Myocardial toxicity was evaluated in neonatal rat myocardiocytes in cultures. In the present study, xanthine oxidase/hypoxanthine(XO/HX) resulted in a decrease in cell viability, increases in LDH activity in culture medium and decreases in beating rate in cultured myocardial cells. In the effect of GHBT extract, it showed the prevention from the XO/HX-induced cardiotoxicity by the increases of cell viability and beating rate as well as the decrease of LDH activity. In the protective effect of Fructus Trichosanthis(FT), Bulbus Allii Macrostemi(BAM) and Rhizoma Pinelliae(RP), all the extracts were significantly effective in the protection of XO/HX-induced cardiotoxocity in cultured myocardial cells by the increase of beating rate as well as th decrease of LDH activity. From these results, they show that XO/HX is cardiotoxic in cultured myocardial cells derived from neonatal rat, and it suggests that GHBT, FT, SAM, RP extracts are positively effective in the blocking in XO/HX-induced cardiotoxicity.

• Nutrition Research and Practice
• /
• v.9 no.6
• /
• pp.592-598
• /
• 2015

BACKGROUND/OBJECTIVES: Increased mass of adipose tissue in obese persons is caused by excessive adipogenesis, which is elaborately controlled by an array of transcription factors. Inhibition of adipogenesis by diverse plant-derived substances has been explored. The aim of the current study was to examine the effects of the aqueous methanol extract of laver (Porphyra yezoensis) on adipogenesis and apoptosis in 3T3-L1 adipocytes and to investigate the mechanism underlying the effect of the laver extract. MATERIALS/METHODS: 3T3-L1 cells were treated with various concentrations of laver extract in differentiation medium. Lipid accumulation, expression of adipogenic proteins, including CCAAT enhancer-binding protein ${\alpha}$, peroxisome proliferator-activated receptor ${\gamma}$, fatty acid binding protein 4, and fatty acid synthase, cell viability, apoptosis, and the total content and the ratio of reduced to oxidized forms of glutathione (GSH/GSSG) were analyzed. RESULTS: Treatment with laver extract resulted in a significant decrease in lipid accumulation in 3T3-L1 adipocytes, which showed correlation with a reduction in expression of adipogenic proteins. Treatment with laver extract also resulted in a decrease in the viability of preadipocytes and an increase in the apoptosis of mature adipocytes. Treatment with laver extract led to exacerbated depletion of cellular glutathione and abolished the transient increase in GSH/GSSG ratio during adipogenesis in 3T3-L1 adipocytes. CONCLUSION: Results of our study demonstrated that treatment with the laver extract caused inhibition of adipogenesis, a decrease in proliferation of preadipocytes, and an increase in the apoptosis of mature adipocytes. It appears that these effects were caused by increasing oxidative stress, as demonstrated by the depletion and oxidation of the cellular glutathione pool in the extract-treated adipocytes. Our results suggest that a prooxidant role of laver extract is associated with its antiadipogenic and proapoptotic effects.

• Environmental Analysis Health and Toxicology
• /
• v.21 no.3 s.54
• /
• pp.245-253
• /
• 2006

We reported previously that glucagon decreased alpha- and pi-class glutathione S-transferases (GSTs) and microsomal epoxide hydrolase (mEN) protein levels in primary cultured rat hepatocytes. The present study examines the effects of Protein kinase A (PKA) inhibitor, KT5720, on the glucagon-mediated decrease in expression of GSTs and mEN. To assess cell viability. lactate dehydrogenase release and MTT activity were examined in hepatocytes treated KT5720. Cell viability was significantly decreased in a concentration dependent manner after incubation with KT5720 at the concentrations of 1 $\mu$M or above for 24 h, which was inhibited by the cytochrome P450 inhibitor SKF-525A. In contrast, another PKA inhibitor H89 (up to 25 $\mu$M) was not toxic to hepatocytes. The glucagon-mediated decrease in expression of alpha- and pi-class GSTs and mEH was completely inhibited by 25 $\mu$M H89 and attenuated by 0.1 $\mu$M KT5720. This study demonstrates that KT5720 may cause cytotoxicity in rat hepatocytes through cytochrome P450-dependent bioactivation. The present study implicates PKA in mediating the inhibitory effect of glucagon on expression of alpha- and pi- class GSTs and mEH.