Ethane 1,2-Dimethane sulfonate(EDS), a toxin which specifically kills Leydig cells(LC), has been widely used to prepare the reversible testosterone(T) depletion rat model. Previous studies including our own clearly demonstrated that the dramatic weight loss of the T-dependent accessory sex organs such as epididymis and seminal vesicle in this 'LC knock-out' rats. These weight loss could be derived from massive and abrupt death of the cells via apoptotic process. The present study was performed to test the effect of EDS administration on the expression of some apoptotic genes in the rat epididymis. Adult male Sprague-Dawley rats($300{\sim}350$ g B.W.) were injected with single dose of EDS(75 mg/kg, i.p.) and sacrificed on Weeks 0, 1, 2, 3, 4, 5, 6 and 7. Tissue weights and the numbers of the epididymal sperm were measured. The transcriptional activities of the bcl-2, bax, Fas and Fas ligand(Fas-L) were evaluated by semi-quantitative RT-PCR. As expected, the weights and the sperm counts of epididymis declined progressively after the EDS treatment during Week 1 and 2. These decrements were discontinued with a gradual return towards normal during Weeks $5{\sim}7$, although the maximal recoveries of the epididymal weights(71%) and sperm count(38%) were subnormal on Week 7. The initial level of bcl-2 transcripts persisted to Week 6 then elevated significantly on Week 7. The level of bax transcripts significantly decreased on Week 6, and no remarkable change was found in the rest of the experimental period. The transcripts for the Fas in epididymis elevated during Weeks $1{\sim}2$, returned to normal on Week 3, and the level persisted to the Week 7. Similarly, the level of Fas-L transcripts elevated during Weeks $1{\sim}3$ and returned to normal after Week 4. Our results demonstrated the transient T depletion by EDS administration could induce the changes in expression of the apoptotic genes in rat epididymis. The activation of Fas and Fas-L in the epididymis of EDS-treated rats might be responsible for the initial apototic process and consequently the tissue damage and the sperm loss. Future studies will attempt to determine the precise molecular mechanism(s) of apoptosis in the rat epididymis.
Kwak, Chung Shil;Kim, Mi-Ju;Kim, Sun Gi;Park, Sunyeong;Kim, In Gyu;Kang, Heun Soo
Journal of Nutrition and Health
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v.52
no.6
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pp.529-539
/
2019
Purpose: Sprouts of evening primrose (Oenothera laciniata, OL) were reported to have high contents of flavonoids and potent antioxidant activity. This study examined the antioxidant and antiobesity activities of OL sprouts to determine if they could be a natural health-beneficial resource preventing obesity and oxidative stress. Methods: OL sprouts were extracted with 50% ethanol, evaporated, and lyophilized (OLE). The in vitro antioxidant activity of OLE was examined using four different tests. The antiobesity activity and in vivo antioxidant activity from OLE consumption were examined using high fat diet-induced obese (DIO) C57BL/6 mice. Results: The IC50 for the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activities of OLE were 26.2 ㎍/mL and 327.6 ㎍/mL, respectively. OLE exhibited the ferric reducing antioxidant power (FRAP) activity of 56.7 ㎍ ascorbic acid eq./mL at 100 ㎍/mL, and an increased glutathione level by 65.1% at 200 ㎍/mL compared to the control in the hUC-MSC stem cells. In an animal study, oral treatment with 50 mg or 100 mg of OLE/kg body weight for 14 weeks reduced the body weight gain, visceral fat content, fat cell size, blood leptin, and triglyceride levels, as well as the atherogenic index compared to the high fat diet control group (HFC) (p < 0.05). The blood malondialdehyde (MDA) level and the catalase and SOD-1 activities in adipose tissue were reduced significantly by the OLE treatment compared to HFC as well (p < 0.05). In epididymal adipose tissue, the OLE treatment reduced the mRNA expression of leptin, PPAR-γ and FAS significantly (p < 0.05) compared to HFC while it increased adiponectin expression (p < 0.05). Conclusion: OLE consumption has potent antioxidant and antiobesity activities via the suppression of oxidative stress and lipogenesis in DIO mice. Therefore, OLE could be a good candidate as a natural resource to develop functional food products that prevent obesity and oxidative stress.
The aim of this study is to know how the rat submandibular gland changes under various emotional stress condition, using molecular biological methods. Restraint and chronic unpredictable mild stress (CUMS) experiment is conducted on fifty one 7-week old Sprague-Dawley rats (restraint stress experiment: 21, CUMS: 30). The rats were sacrificed, the submandibular glands were excised immediately at certain time, and examined by the use of immunohistochemistry and western blotting. In CUMS experiment, sucrose preference test, water intake change, weight change were implemented at 1 week interval for the experimental period The results are as follows: 1. The number of clusterin-secreting cells of restraint stress group compared to control group showed significantly decreasing tendency in all experimental groups except for the 1st hour group (p<0.001 in the 9th, 24th, 72nd, 120th, and 168th hour group). 2. The number of clusterin-secreting cells of CUMS group compared to control group showed significantly increasing tendency in the 2nd week group (p<0.01), and significantly decreasing tendency in the 4th and 5th week group (p<0.001). 3. Sucrose preference test in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01). 4. Weight change in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01), but water intake change didn't show significant difference compared to control group. 5. In western blot analysis, clusterin expression was decreased on a gradual basis in due time compared to the control group in the restraint stress group. As for CUMS group (chronic unpredictable mild stress group), it was increased till the 2nd week and decreased till the 5th week after that, which is similar to immunohistochemical analysis result and the decreasing tendency of sucrose preference and weigh changes. Through the test, it was proved that expression of clusterin in saliva glands decreases after receiving either acute or chronic stress, indicating relation with depression caused by chronic stress. Unlike other data, however, apoptotic tendency was hardly found in tissues. Diverse possibilities could be suggested on that: first, the stress was not enough to expedite apoptosis; second, apoptosis-related protein was already being secreted though not detected with microscope; third, clusterin, a major secretion molecule of saliva, decreased with saliva's malfunction due to stress. In the respect, it will be necessary to examine proteins expressed in case of cell death or other heat-shock proteins at the same time, in order to see whether any cellular change or death is caused by decreasing clusterin under high stress, and whether the original state is restored as time goes by under mild stress, through longer-term tests using even higher acute stress.
Thi, Luc The;Nguyen, Quan Hoang;Park, Yoo Gyeong;Jeong, Byoung Ryong
Journal of Bio-Environment Control
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v.28
no.2
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pp.178-184
/
2019
Strawberry ($Fragaria{\times}ananassa$) is one of the most important and popular fruit crops in the world, and 'Sulhyang' is one of the principal cultivars cultivated in the Republic of Korea for the domestic market. The growth and flower induction in strawberry is the process which influences directly on fruit bearing and yield of this crop. In this study, effect of benzyladenine (BA), gibberellic acid ($GA_3$), and salicylic acid (SA) on growth and flower bud induction in strawberry 'Sulhyang' was investigated. The 3-week-old runner plants, grown in 21-cell propagation trays, were potted and cultivated in growth chambers with $25^{\circ}C/15^{\circ}C$ (day/night) temperatures, 70% relative humidity (RH), and light intensity of $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ photosynthetic photon flux density (PPFD) provided by white light emitting diodes (LEDs). The runner plants were treated with one of three concentrations, 0 (control), 100, and $200mg{\cdot}L^{-1}$ of BA, $GA_3$, or SA solution. The chemicals were sprayed two times on leaves of runner plants at an interval of two weeks. After 9 weeks the results showed that the application of all chemicals caused reduction of root length and chlorophyll (SPAD) content as compared to the control. The lowest chlorophyll (SPAD) content was recorded in plants treated with $GA_3$. However, the treatment of $200mg{\cdot}L^{-1}$$GA_3$ promoted leaf area, leaf fresh weight, and plant fresh weight. The greatest flower induction (85%) and number of inflorescences (4.3 inflorescences per plant) were observed in the treatment of $200mg{\cdot}L^{-1}\;SA$, followed by $100mg{\cdot}L^{-1}\;SA$. Overall, results suggest that foliar application of $GA_3$ solution could accelerate plant growth, while foliar application of SA solution could induce hastened flowering. Further studies may be needed to find out the relationship between $GA_3$ and SA solutions treated in a combination, and the molecular mechanism involved in those responses observed.
Hwang-Bo, Hyun;Kwon, Da Hye;Kim, Min Young;Ji, Seon Yeong;Choi, Eun Ok;Kim, Sung Ok;Jeong, Ji-Suk;Hong, Su Hyun;Choi, Sung Hyun;Park, Cheol;Choi, Yung Hyun
Journal of Life Science
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v.29
no.1
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pp.112-117
/
2019
Herbal medicines are widely used as therapeutic products in many countries. Corni fructus (CF), the dried ripe sarcocarp of Cornus officinalis Sieb. et Zucc (Cornaceae), has been used for thousands of years in traditional medicine and has been reported to be effective for the prevention and treatment of various diseases, such as kidney diseases and diabetes. Recent research on CF has documented a wide spectrum of therapeutic properties, which include anti-inflammatory, ant-oxidative, immunomodulatory, and anti-cancer effects. However, there is no information on its safety. Therefore, in this study, the toxicity of water extract of CF to ICR mice was investigated. The mice received a single dose of water extract of CF (1,000, 2,000, and 5,000 mg/kg of body weight) via the oral route. Mortality, clinical signs, body weight changes, gross findings, and weights of the principal organs after 14 d were then assessed. The results revealed no adverse effects of CF as determined by clinical signs, body weights, or organ weights and no gross pathological findings in any of the treatment groups. These results suggest that the 50% lethal dose and approximated lethal dose of CF extract is over 5,000 mg/kg. The findings provide scientific evidence for the safety of CFs.
Sucrose (suc) is a disaccharide that consists of glucose (glu) and fructose (fru). It is a carbohydrate source that acts as a nutrient molecule and a molecular signal that regulates gene expression and alters metabolites. This study aimed to evaluate whether suc-specific signaling induces an increase in bioactive compounds by exogenous suc absorption via roots or whether other factors, such as osmotic stress or biotic stress, are involved. To compare the osmotic stress induced by suc treatment, 4-week-old cultured mugwort plants were subjected to Hoagland nutrient solution with 10 mM, 30 mM, and 50 mM of suc or mannitol (man) for 3 days. Shoot fresh weight in suc and man treatments was not significantly different from the control. Both man and suc treatments increased the content of bioactive compounds in mugwort, but they displayed different enhancement patterns compared to the suc treatments. Mugwort extract treated with suc 50 mM effectively protected HepG2 liver cells damaged by ethanol and t-BHP. To compare the biotic stress induced by suc treatment, 3-week-old mugwort plants were subjected to microorganism and/or suc 30 mM with Hoagland nutrient solution. Microorganisms and/or suc 30 mM treatments showed no difference about the shoot fresh weight. However, sugar content in mugwort treated with suc 30 mM and microorganism with suc 30 mM treatment was significantly higher than that of the control. Suc 30 mM and microorganism with suc 30 mM were effective in enhancing bioactive compounds than microorganism treatment. These results suggest that mugwort plants can absorb exogenous suc via roots and the enhancement of bioactive compounds by suc treatment may result not from osmotic stress or biotic stress because of microorganism, but by suc-specific signaling.
The apple seed contained 25.96% of crude fat and 37.62% of crude protein. The lipid fractions obtained by cilicic column chromatography were mainly composed of about 93.52% neutral lipid, whereas compound lipid was only 6.48% level. Among the neutral lipid separated by thin layer chromatography, triglyceride was 92.17%, sterol ester, sterol, diglyceride and free fatty acid were 3.53, 2.25, 1.44 and 0.56, respectively. The predominent fatty acids of total and neutral lipids were linoleic acid (59.79-69.37%) and oleic acid (20.04-29.82%), but those of glycolipid and phojspholipid were linoleic acid (29.20-36.04%). The major fatty acids of triglyceride separated from neutral lipid were oleic acid (44.31%), linoleic acid (36.66%) and palmitic acid (12.48%). The salt soluble protein of apple seed was highly dispersible in 0.02M sodium phosphate buffer containing about 1.0M $MgSO_4$, and the extractability of seed protein was 37%, Glutamic acid was the major amino acid in salt soluble protein, followed by arginine and aspartic acid. The eletrophoretic analysis showed three bands in apple seed protein, and the collection rate of the main protein fraction purified by Sephadex G-100 and G-200 was 76.6%. Glutamic acid, aspartic acid and arginine were the major amino acids of the main apple seed protein. The molecular weight for the main protein of the apple seed was estimated to be 45,000.
Fish sauces prepared from sardine, Sardinops melanosticta were tested for inhibitory activity against angiotensin-I converting enzyme(ACE). Three kinds of fish sauces were prepared from scrap(S), meat(M) and round(R) of sardine, respectively. ACE inhibitory activity of sardine sauce S and R decreased with the elapse of fermentation period, whereas that of sardine sauce M increased to 30 days and thereafter decreased. ACE inhibitory activity of sardine sauce M fermented with koji was higher than that without koji. And occurrence of $5\%$ TCA soluble peptide-nitrogen was similar to tendancy of the ACE inhibitory activity. The ACE inhibitory activity increased with an increment of amounts added and was stable at heat treatment in boiling water bath for 5hrs. $IC_{50}\%$ (Amounts of inhibitors need for $50\%$ inhibition) of the sardine sauce S, M and R fermented with(without) koji during 90 days was $125{\mu}g(140{\mu}g),\;200{\mu}g(100{\mu}g)$ and $125{\mu}g(135{\mu}g)$, respectively. From the profiles of fractionation of the sardine sauce R fermented without koji for 90 days, the molecular weight of most active fraction was about 1,400 and the amino acids of Glu, Ala, Leu and Lys were found in abundance.
Previous biochemical studies indicate that $(Na^++K^+)-ATPase$ is composed of two subunits, ${\alpha}$ and ${\beta}$, in a form of ${\alpha}_2{\beta}_2$ with a molecular weight of approximately 300,000 daltons. There is also suggestive evidence that the $Na^+$, $K^+$ pump in human erythrocytes occurs in a complex with some glycolytic enzymes. We assessed here in situ assembly size of the $(Na^++K^+)-ATPase$ of human erythrocytes by applying classical target theory to radiation inactivation data of the ouabain-sensitive sodium flux and ATP hydrolysis of intact cells and ghosts. Cells(in the presence of cryoprotective agent) and ghosts were irradiated at $-45^{\circ}C$ to $-50^{\circ}C$ with an increasing dose of a 1.5 MeV electron beam, and after thawing, the pump and/or enzyme activities were assayed. Each activity measured was decreased as a simple exponential function of radiation dose, from which a radiation sensitive volume (target size) was calculated. When intact cells were used, the target size of both $(Na^++K^+)-ATPase$ and $Na^+$, $K^+$ pump was found to be approximately 600,000 daltons. This target size of the ATPase was reduced to approximately 325,000 daltons if the cells were pretreated with strophanthidin. When ghosts were used, the target size of the ATPase was again approximately 325,000 daltons. Our target size measurement suggests that, in intact cells, the $(Na^++K^+)-ATPase/Na^+,K^+$ pump exists either as a dimer of $(\alpha\beta)_2$ which is a functional unit or as a monomer of $(\alpha\beta)_2$ but in tight complex with other enzyme or enzymes. The results also suggest that this dimeric or heterocomplex association is dissociated during ghost preparation and strophanthidin treatment.
Lee, Ho Se;Jeon, Sang Goo;Oh, You-Kwan;Kim, Kwang Ho;Chung, Soo Hyun;Na, Jeong-Geol;Yeo, Sang-Do
Korean Chemical Engineering Research
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v.50
no.4
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pp.672-677
/
2012
Lipids in microalgal biomass were recovered by using pyrolysis method. The pyrolysis experiments of two Chlorella sp. KR-1 samples, which have triglyceride contents of 10.8% and 36.5%, respectively were carried out at $600^{\circ}C$ to investigate the effects of lipid contents in the cells on the reaction characteristics. The conversion and liquid yield of the lipid-rich sample were higher than those of the lipid-lean sample since its carbon to hydrogen ratio was low. There were low molecular weight organic acids, ketones, aldehydes and alcohols in the liquid products from both KR-1 samples, but the pyrolysis oil of the lipid-rich sample was abundant in free fatty acids, particularly palmitic acid, oleic acid and stearic acid while the content of nitrogen containing organic compounds was low. The microalgal pyrolysis oil had two layers composed of the light hydrophobic fraction and the heavy hydrophilic fraction. The light fraction might be originated from triglycerides and the heavy fraction might be from carbohydrates and proteins. In the light fraction of the liquid products, there were considerable linear alkanes such as pentadecane and heptadecane as well as free fatty acids, implying that deoxygenation reaction including decarboxylation was occurred during the pyrolysis. The yield of the liquid products from the pyrolysis of the KR-1 sample having triglyceride content of 36.5% was 56.9% and the light fraction in the liquid products was 68.2%. Also more than 80% of the light fraction was free fatty acids and pure hydrocarbons, thus showing that most triglycerides could be extracted in the form of suitable raw materials for biofuels.
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