Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.
Park, Mi Hee;Kim, Chul Jin;Lee, Jin Young;Keum, Chang Yeop;Kim, In Seon;Jin, Chang Hyun;Ji, Joong-Gu;Kim, Sung-kyu
Journal of the Korean Applied Science and Technology
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v.37
no.1
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pp.102-113
/
2020
In this study, anti-arthritic effect of the mixed extract of radiation mutant Perilla frutescens var. crispa and Atractylodes macrophala koidz was investigated. Cell viability was determined by MTT assay in RAW 264.7 cells. The anti-inflammatory effect of mixed extracts was determined through measurement of the levels of reactive oxygen species (ROS) and nitric oxide (NO), release of inflammatory cytokines and expression of NF-κB, COX-2 and iNOS in LPS-induced RAW 264.7 cells after treatment of mixed extracts (5, 10, 25 ㎍/㎖). We showed that the mixed extracts was not toxic in the dose of 5, 10, 25 ug/ml, and significantly inhibited production of nitric oxide and ROS, cytokines including IL-1β, IL-6, TNF-α, and inflammatory proteins including NF-κB, COX-2 and iNOS in LPS-induced RAW 264.7 cells. Moreover, the mixed extract inhibited the type II collagen induced arthritis in DBA mice in the dose of 66.5 and 133mg/kg/day. Therefore, we suggest that mixed extract of radiation mutant Perilla frutescens var. crispa and Atractylodes macrophala koidz can be developed as a raw material for health functional food and therapeutics to treat the inflammatory arthritis.
Journal of the Korean Society for Marine Environment & Energy
/
v.18
no.2
/
pp.51-63
/
2015
In order to investigation long-term variations of water qualities in the Saemangeum Salt-Water Lake formed after the sea-dike construction, the survey has carried out over 40 time from 2002 to 2010. The decreased salinity in surface water immediately after the dike construction has maintained on equal terms for years. After the dike construction, the early concentration of SPM in surface water has decreased but then it showed the tendency to move up and down due to the changes of water level in the lake. The elevated concentration of Chl-a in surface water initially after the dike construction was kept at the same conditions for years. The concentration of DIN in surface water has not changed before and shortly after the dike construction. However, the concentration of $NH_4-N$ in surface water has increased steadily after the dike construction. Consequently the concentration of DIN in the lake water after years has raised compared to pre-dike construction. The reduced concentration of DIP in surface water soon after the dike construction has increased after years as well as $NH_4-N$ due to the accumulation of organic matter to inside lake. Unlike with the unvaried $NO_3-N$, the concentration of DISi in surface water after the dike construction has immediately increased and maintained the enhanced level indicating the supply from other sources except the freshwater. Since the dike construction, the spatial characteristics of water quality was divided river sides and rest of the lake markedly. Stratification of river sides was more strong than the dike sides. In the warm seasons, hypoxia causing the release of nutrients and metals from sediment was observed downward about 1 m from surface of river sides. We strongly suggest to make some urgent measure to prevent low dissolved oxygen condition in the bottom layer of the river sides.
Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Park, Sang Hyun;Park, Su Bin;Yoo, Seul Ki;Han, Hye Ju;Lee, Su-Gwang;Lee, Uk;Heo, Ho Jin
Korean Journal of Food Science and Technology
/
v.50
no.2
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pp.225-237
/
2018
This study was conducted to compare nutritional analysis and neuroprotective effect of 5 cultivars of Diospyros kaki (Dungsi, Godongsi, Gojongsi, Gabjubaekmok, and Bansi). In nutritional analysis, three free sugars: sucrose, glucose, and fructose, and six fatty acids: tartaric acid, hexadecanoic acid, palmitic acid, oleic acid, octadecenamide, and octadecane, were detected. Potassium and phosphorus levels were the highest in inorganic component analysis, and glutamic acid and aspartic acid were the highest contents in amino acid analysis. Vitamin C was detected in all cultivars. Total phenolic content was the highest in Dungsi. Antioxidant activities such as ABTS (3-ethylbenzothiazoline-6-sulfonic acid), DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activities, FRAP (ferric reducing/antioxidant power), and MDA (malondialdehyde) inhibitory effect were the highest in Gabjubaekmok. Acetylcholinesterase inhibitory activity, cell viability, intracellular reactive oxygen species (ROS) accumulation, and lactate dehydrogenase (LDH) release were measured to confirm the neuroprotective effect in MC-IXC cells. Gabjubaekmok showed significant acetylcholinesterase (AChE) inhibition and neuroprotection.
Wetlands constitute a transitional zone between terrestrial and aquatic ecosystems and have unique characteristics such as frequent inundation, inflow of nutrients from terrestrial ecosystems, presence of plants adapted to grow in water, and soil that is occasionally oxygen deficient due to saturation. These characteristics and the presence of vegetation determine physical and chemical properties that affect decomposition rates of organic matter (OM). Decomposition of OM is associated with activities of various extracellular enzymes (EE) produced by bacteria and fungi. Extracellular enzymes convert macromolecules to simple compounds such as labile organic carbon (C), nitrogen (N), phosphorus (P), and sulfur (S) that can be easily taken up by microbes and plants. Therefore, the enzymatic approach is helpful to understand the decomposition rates of OM and nutrient cycling in wetland soils. This paper reviews the physical and biogeochemical factors that regulate extracellular enzyme activities (EEa) in wetland soils, including those of ${\beta}$-glucosidase, ${\beta}$-N-acetylglucosaminidase, phosphatase, arylsulfatase, and phenol oxidase that decompose organic matter and release C, N, P, and S nutrients for microbial and plant growths. Effects of pH, water table, and particle size of OM on EEa were not significantly different among sites, whereas the influence of temperature on EEa varied depending on microbial acclimation to extreme temperatures. Addition of C, N, or P affected EEa differently depending on the nutrient state, C:N ratio, limiting factors, and types of enzymes of wetland soils. Substrate quality influenced EEa more significantly than did other factors. Also, drainage of wetland and increased temperature due to global climate change can stimulate phenol oxidase activity, and anthropogenic N deposition can enhance the hydrolytic EEa; these effects increase OM decomposition rates and emissions of $CO_2$ and $CH_4$ from wetland systems. The researches on the relationship between microbial structures and EE functions, and environmental factors controlling EEa can be helpful to manipulate wetland ecosystems for treating pollutants and to monitor wetland ecosystem services.
Jin, Su Il;Kim, Hyeon Ju;Jeong, Ji Hee;Jin, Dong Eun;Choi, Sung-Gil;Heo, Ho Jin
Korean Journal of Food Science and Technology
/
v.46
no.5
/
pp.609-615
/
2014
The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.
The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.
Ko, Sun Young;Kang, Saem;Chang, Yun Sil;Park, Eun Ae;Park, Won Soon
Clinical and Experimental Pediatrics
/
v.49
no.3
/
pp.317-325
/
2006
Purpose : This study was carried out to elucidate the effects of nitric oxide synthase(NOS) inhibitor, NG-monomethyl-L-arginine(L-NMMA) and nitric oxide precursor, L-arginine(L-Arg) on cerebral hemodynamics and energy metabolism during reoxygenation-reperfusion(RR) after hypoxia-ischemia(HI) in newborn piglets. Methods : Twenty-eight newborn piglets were divided into 4 groups; Sham normal control(NC), experimental control(EC), L-NMMA(HI & RR with L-NMMA), and L-Arg(HI & RR with L-Arg) groups. HI was induced by occlusion of bilateral common carotid arteries and simultaneously breathing with 8 percent oxygen for 30 mins, and followed RR by release of carotid occlusion and normoxic ventilation for one hour. All groups were monitored with cerebral hemodynamics and cytochrome $aa_3$ (Cyt $aa_3$) using near infrared spectroscopy(NIRS). $Na^+$, $K^+$-ATPase activity, lipid peroxidation products, and tissue high energy phosphate levels were determined biochemically in the cerebral cortex. Results : In experimental groups, mean arterial blood pressure, $PaO_2$, and pH decreased, and base excess and blood lactate level increased after HI compared to NC group(P<0.05). These variables subsequently returned to baseline after RR except pH. There were no differences among the experimental groups. In NIRS, oxidized hemoglobin($HbO_2$) decreased and hemoglobin(Hb) increased during HI(P<0.05) but returned to base line immediately after RR; 40 min after RR, the $HbO_2$ had decreased significantly compared to NC group(P<0.05). Changes of Cyt $aa_3$ decreased significantly compared to NC after HI and recovered at the end of the experiment. Significantly reduced cerebral cortical cell membrane $Na^+$, $K^+$-ATPase activity and increased lipid peroxidation products(P<0.05) were not improved with L-NMMA or L-Arg. Conclusion : These findings suggest that NO is not involved in the mechanism of HI and RR brain damage during the early acute phase of RR.
In, Kwang-Ho;Lee, Jin-Goo;Cho, Jae-Youn;Shim, Jae-Jung;Kang, Kyung-Ho;Yoo, Se-Hwa
Tuberculosis and Respiratory Diseases
/
v.41
no.3
/
pp.231-238
/
1994
Backgroud: Since the demonstration of the fact that vascular relaxation by acetylcholine(Ach) results from the release of relaxing factor from the endothelium, the identity and physiology of this endothelium-derived relaxing factor(EDRF) has been the target for many researches. EDRF has been identified as nitric oxide(NO). With the recent evidences that EDRF is an important mediator of vascular tone, there have been increasing interests in defining the role of the EDRF as a potential mediator of hypoxic pulmonary vasoconstriction. But the role of EDRF in modulating the pulmonary circulation is not compeletely clarified. To investigate the endothelium-dependent pulmonary vasodilation and the role of EDRF during hypoxic pulmonary vasoconstriction, we studied the effects of $N^G$-monomethyl-L-arginine(L-NMMA) and L-arginine on the precontracted pulmonary arterial rings of the rat in normoxia and hypoxia. Mothods: The pulmonary arteries of male Sprague Dawley(300~350g) were dissected free of surrounding tissue, and cut into rings. Rings were mounted over fine rigid wires, in organ chambers filled with 20ml of Krebs solution bubbled with 95 percent oxygen and 5 percent carbon dioxide and maintained at $37^{\circ}C$. Changes in isometric tension were recorded with a force transducer(FT.03 Grass, Quincy, USA) Results: 1) Precontraction of rat pulmonry artery with intact endothelium by phenylephrine(PE, $10^{-6}M$) was relaxed completely by acetylcholine(Ach, $10^{-9}-10^{-5}M$) and sodium nitroprusside(SN, $10^{-9}-10^{-5}M$), but relaxing response by Ach in rat pulmonary artery with denuded endothelium was significantly decreased. 2) L-NMMA($10^{-4}M$) pretreatment inhibited Ach($10^{-9}-10^{-5}M$)-induced relaxation, but L-NMMA ($10^{-4}M$) had no effect on relaxation induced by SN($10^{-9}-10^{-5}M$). 3) Pretreatment of the L-arginine($10^{-4}M$) significantly reversed the inhibition of the Ach ($10^{-9}-10^{-5}M$)-induced relaxation caused by L-NMMA($10^{-4}M$) 4) Pulmonary arterial contraction by PE($10^{-6}M$) was stronger in hypoxia than normoxia but relaxing response by Ach($10^{-9}-10^{-5}M$) was decreased, 5) With pretreatment of L-arginine($10^{-4}M$), pulmonary arterial relaxation by Ach($10^{-9}-10^{-5}M$) in hypoxia was reversed to the level of relaxation in normoxia. Conclusion: It is concluded that rat pulmonary arterial relaxation by Ach is dependent on the intact endothelium and is largely mediated by NO. Acute hypoxic pulmonary vasoconstriction is related to the suppression on NO formation in the vascular endothelium.
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