• Toxicological Research
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• 제18권4호
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• pp.355-362
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• 2002

Pathophysiological elevation of intracellular calcium concentration ($[Ca^{2+}]_1$) in the neuron has been considered as an important responsible factor in the neuronal cell damages. However the mechanism of increase of $[Ca^{2+}]_1$ and the relationship between $[Ca^{2+}]_1$ level and cytotocixity have not been fully demonstrated. In the present study, real-time alteration of $[Ca^{2+}]_1$and cellular response (cell damages) in the pheochromocytoma cells (PC12) stimulated by glutamate were investigated. Glutamate dose dependently decreased cell viability determined propidium iodide fluorescence method and morphology change. Conversely related with cell damages, glutamate dose dependently increased the level of［Ca$^{2+}$］$_{i}$ . To investigate the mechanism of glutamate-induced increase of $[Ca^{2+}]_1$,$[Ca^{2+}]_1$, was first measured in the cell cultured in calcium free media and in the presence of dantrolene, an inhibitor of calcium release from ryanodine receptor located in endoplasmic reticulum (ER). Similar to the increase$[Ca^{2+}]_1$ in the calcium-containing media, glutamate dose dependently increased $[Ca^{2+}]_1$ in the cell cultured in free calcium media. However pretreatment (2 hr) with 20~50 $\mu\textrm{M}$ dantrolene substantial lowered glutamate-induced increase of $[Ca^{2+}]_1$, suggesting that release of calcium from ER may be major sourse of increase of $[Ca^{2+}]_1$ in PC12 cells. Dantrolene-induced inhibition of $[Ca^{2+}]_1$ resulted in recovery of cytotoxicity by glutamate. Relevance of N-methy-D-aspartate (NMDA) receptor, a type of glutamte receptor on glutamate-induced incense of $[Ca^{2+}]_1$,$[Ca^{2+}]_1$ was also determined in the cells pretreated (2 hr) with NMDA receptor antagonist MK-80l. Glutamate-induced increase of $[Ca^{2+}]_1$ was reduced by MK-801 dose dependently. Furthermore, glutamate-induced cytotoxicity was also prevented by MK-80l. These results demonstrate that glutamte increase $[Ca^{2+}]_1$ dose dependently and thereby cause cytotoxicity. The increase of $[Ca^{2+}]_1$ may release from ER, especially through ryanodine receptor and/or through NMDA receptor Alteration of calcium homeostasis through disturbance of ER system and/or calcium influx through NMDA receptor could contribute glutamate-induced cell damages.s.

• 대한미생물학회지
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• 제35권3호
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• pp.251-261
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• 2000

V. vulnificus is an estuarine bacterium which causes septicemia and shock in susceptible patients. The organism produces a hemolytic cytolysin (VvH), which has a membrane damaging effect on erythrocytes. To clarify the mechanisms by which VvH might contribute to virulence, we examined its effect on macrophages. When mouse peritoneal macrophages were harvested and co-cultured with hemolysin-positive V. vulnificus strains (100 bacteria/cell), about 60% of the macrophages were killed; macrophages were not killed when co-cultured V. vulnificus strain CVD 707, a VvH-negative deletion mutant. Exposure of macrophages to filtered culture supernatants (2.5 HU/ml) and purified VvH (3 HU/ml) resulted in an increase in dead cells (80 and 90%, respectively), as determined by the trypan blue dye exclusion method and LDH release from macrophages was also increased (70 and 65.5%, respectively). The cytotoxic effect of VvH on macrophages was both the dose- and time-dependent. The VvH caused damage to the macrophage membrane and was blocked significantly by preincubation with cholesterol (p<0.01). Fetal bovine serum showed remarkable inhibition of VvH synthesis by V. vulnificus and inhibited VvH activity in culture supernatant. Cell viability was increased by 35% (p<0.01) and LDH release decreased by 28% (p<0.01) when macrophages were incubated with V. vulnificus (100 bacterial cell) in DMEM-10% FBS for 2 hr. Bacterial clearance activity of mice against V. vulnificus CVD 707 was decreased by pretreatment with 10 HU of VvH. This result suggests that the VvH can impair the membrane of macrophages and may playa role in the pathogenesis of V. vulnificus septicemia.

• Journal of Radiation Protection and Research
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• 제24권2호
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• pp.93-99
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• 1999

Alkaline single cell gel electrophoresis (SCGE) assay는 일명 혜성분석이라고 부르며 in vivo 와 in vitro 에서 많은 화학적, 생물학적인 인자에 의한 DNA 손상을 감지하는데 유용한 기법으로 각각의 세포에서 DNA 단일 가닥 절단과 알칼리에 약한 장소를 평가하는 새로운 방법으로 인정되고 있다. 단세포 겔 전기영동법 (SCGE)을 사용하여 복숭아씨 추출물이 방사선에 의하여 사람 림프구 DNA에 나타나는 손상을 보호하는 지 여부를 평가하였다. 복숭아씨 추출물로 10 분간 전처리한 림프구를 0, 0.1, 0.3, 0.5, 1.0, 2.0 Gy 의 방사선으로 조사하였고 방사선만을 조사한 림프구 실험군과 비교평가하였다. 혜성분석에서 DNA 가닥 절단에 대한 표식인 tail moment의 증가는 감마선에 대해서 뚜렷한 선량-반응 관계를 나타내었으며 각각의 농도별로 복숭아씨 추출물이 처리된 림프구의 DNA 손상은 현저히 감소하였다. 단세포 겔 전기영동법을 통한 평가결과 복숭아씨 추출물은 방사선에 의한 림프구 DNA 손상에 대한 탁월한 방어효과를 나타내었다.

• Restorative Dentistry and Endodontics
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• 제35권3호
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• pp.173-179
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• 2010

본 연구의 목적은 실란, 과산화수소, 불산, 샌드블라스팅을 이용한 섬유포스트의 표면 처리가 복합레진 코어와의 결합강도에 미치는 영향을 평가하기 위한 것이다. 32개의 FRC Postec Plus를 표면 처리를 하지 않은 대조군과 표면 처리 방법에 따라 7개의 실험군(S군, HP군, HP-S군, HF군, HF-S군, SB군, SB-S군)으로 분류하였다. S군은 실란을 적용하였고, HP군은 28%의 과산화수소로 부식하였으며, HP-S군은 과산화수소 처리 후 실란을 적용하였다. HF군은 4% 불화수소산 젤로 부식하였고, HF-S군은 불화수소산 젤 처리 후 실란으로 처리하였다. SB군은 산화 알루미늄으로 샌드블라스팅 하였고, SB-S군은 샌드블라스팅 후 실란을 적용하였다. 표면 처리한 포스트는 Tetric Flow를 사용하여 광중합 시키면서 주위에 원통형의 코어를 축조하였다. 형성된 포스트-코어 복합체를 막대 형태로 절단하여 미세인장결합강도를 측정하였다. One-way ANOVA와 LSD test로 결합강도 값을 분석한 결과, 샌드블라스팅은 섬유포스트와 복합레진 코어와의 결합력을 증가시켰으며 샌드블라스팅 후 실란으로 처리한 경우 다른 처리 방법들보다 유의하게 증가된 결합력을 나타냈다(p < 0.05).

• 대한한방소아과학회지
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• 제20권1호
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• pp.241-255
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• 2006

Objective : Allergic Inflammation is related with secretion of Cytokine. This study was performed to examine the effects of Woobangja on anti-allergic inflammation. Method : While macrophage 264.7cells was chosen as a normal group a control group was classified into three groups. One was stimulated with LPS. and another was pretreated with Woobangja for 1 hour. The third was pretreated with gydrocortisone for 1 hour. After the pretreatment, macrophage were incubated with lipopolysaccharide(LPS) 100 ng/ml for 12h and media collected and $TNF-{\alpha}$, IL-6, $IL-1{\beta}$, IL-10 concentration in supernatants were measured each by Enzyme linked immuno-sorbent assay. Woobangja were used $50\;{\mu}g/ml$, $100\;{\mu}g/ml$, $250\;{\mu}g/ml$, $500\;{\mu}g/ml$, 1 mg/ml. Hydrocortisones were used respectively $10^{-8}\;M$,$10^{-7}\;M$,$10^{-6}\;M$,$10^{-5}\;M$,$10^{-4}\;M$. Results : Woobangja showed inhibitory effect on $TNF-{\alpha}$ by LPS-stimulated macrophage 264.7. The inhibitory effect was most significant in 1mg/ml(p<0.01), and has increased according to the number of doses. Woobangja also showed inhibitory effect on IL-10 by LPS-stimulated macrophasg 264.7. The inhibitory effect was most significant in $100\;{\mu}g/ml$, and was not in a dose-dependent manner as Hydrocortisone group. Woobangja and Hydrocortison showed contrary effect on $IL-1{\beta}$ in al five concentration(p<0.01), and at the lowest concentration ($50\;{\mu}g/ml$) the level of $IL-1{\beta}$ was the lowest. On the other hand hydrocortison was observed to have inhibitory effect on $IL-1{\beta}$ in all five concentration(p<0.01). IL-6 was inhibited by hydrocortison in a roughly dose-dependent manner, but was not inhibited by Woobangja. On the contrary Woobangja obviously increased the expression of $IL-1{\beta}$ in all five concentration(p<0.01), but it was not related with concentrations. Conclusion : 1. Woobangja does significantly inhibit the expression of $TNF-{\alpha}$ by LPS-stimulated macrophage 264.7. 2. Woobangja does significantly increse the expression of IL-6 by LPS-stimulated macrophage 264.7. 3. Woobangja does significantly increse the expression of $IL-1{\beta}$ by LPS-stimulated macrophage 264.7. 4. Woobangja does significantly inhibit the expression of IL-10 by LPS-stimulated macrophage 264.7. 5. Woobangja is observer to have anti-allergic inflammatory effect through inhibiting inflammatory cytokine.

• 대한한방내과학회지
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• 제25권4호
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• pp.289-299
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• 2004

Background : Although the pathophysiology of asthma has been reported, its mechanism has not been fully elucidated. The mast cell is an effector cells in allergic inflammation and secretes a number of chemokines. Chemokines are important for the recruitment of leukocytes to sites of infection, which is essential in host defense. Chemokines also contribute to the pathogenesis of several disorders such as asthma, chronic bronchitis, atopic dermatitis, allergic rhinitis, and rheumatoid arthritis. Objective : In this study, the aim was to identify the effect of Baekryunchihyo-tang(白蓮治哮湯) on expression of chemokines. This was examined by RT-PCR using the human mast cell line (HMC-l) Materials and Methods : HMC-l cells were used, which is known to secrete and express chemokines. In order to investigate the protective effect of Baekryunchihyo-tang(白蓮治哮湯), HMC-l cells were incubated with pretreatment of Baekryunchihyo-tang(白蓮治哮湯) for 24 hrs. RT-PCR analyses of chemokine genes of cells pretreated with Baekryunchihyo-tang(白蓮治哮湯) showed that expressions of IL-8, $MIP-l{\beta}$, and RANTES genes in these cells were lower and $MIP-l{\alpha}$ showed a similar pattern compared to the calcium ionophore-treated group. In addition, cell cytotoxicity concentration measurements were performed by MTT assay method. Results : After stimulation with 1 uM calcium ionophore A23178 for 2 hrs, IL-8, major one of CXC chemokines, was highly expressed, and expression of $MIP-l{\beta}$ and RANTES (CC chemokines) increased, while expression of $MIP-l{\alpha}$ did not change. The cell cytotoxicity of Baekryunchihyo-tang(白蓮治哮湯) with treatments at various concentrations and times was not observed, respectively. Conclusion : This study suggests that Baekryunchihyo-tang(白蓮治哮湯) has dose-dependent effects on mRNA expression of IL-8(CXC chemokines), $MIP-l{\beta}$ and RANTES(CC chemokines) in human mast cellline(HMC-l). So these herbal medicines may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanism of inhibition by herbal medicine in the asthma model. This study provides basic data on the possibility of the clinical treatment of Baekryunchihyo-tang(白蓮治哮湯) for allergic disorders.

• 한국식품과학회지
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• 제45권3호
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• pp.312-316
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• 2013

본 연구에서는 탕액에 존재하는 제랄레논의 분석을 위한 HPLC법을 확립하고 탕액조제 시 한약재로부터의 제랄레논 이행률을 조사하였다. 한약재 (괄루인, 두충, 복분자)에 제랄레논을 임의의 농도로 오염시켜 수침 및 비수침 과정을 거친 후 상압가열 ($100^{\circ}C$, 3 h)과 고압가열 ($121^{\circ}C$, 1 h)하여 탕액을 조제한 다음 탕액과 한약재 잔류물로 분리하고 immunoaffinity column으로 정제하여 분석에 사용하였다. 가열처리를 하지 않은 탕액 제조 전 원료 한약재에 대한 회수율은 괄루인 83.7-95.5%, 두충 81.9-99.7%, 복분자 79.1-82.3%로 분석에 이용이 가능한 것으로 확인되었고, 3종의 한약재 중 회수율이 가장 좋은 두충을 선택하여 탕액을 조제한 후 탕액과 한약재 잔류물에 대한 회수율 측정한 결과 탕액에서는 68.4-83.7%, 한약재 잔류물에서는 72.9-80.3%로 원재료 보다는 낮은 수준으로 확인되었다. 한약재에서 탕액으로 제랄레논의 이행률은 한약재 원재료에 제랄레논을 임의의 농도로 오염시킨 후 탕액을 조제하여 분석하였으며, 그 결과 탕액에서는 제랄레논이 검출되지 않았고, 괄루인과 복분자 잔류물에서는 수침 시료의 경우 17.04-22.99%, 비수침 시료의 경우 10.17-18.33%의 제랄레논이 검출되었다. 그리고 두충 잔류물에서는 수침 시료의 경우 10.44-17.16%, 비수침 시료의 경우 12.42-17.75%가 검출되었다. 본 연구에서는 탕액에서 제랄레논이 검출되지 않아 이행률은 낮은 것으로 판단되나 보다 탕액 조제과정 중 첨가물에 의해 이행이 일어날 수 있는 가능성은 여전히 존재하기 때문에, 제랄레논을 포함한 곰팡이독소에 안전한 한약재의 확보를 위한 지속적인 노력이 필요하다. 또한, 유통 중인 한약재를 포함하여 조제포장되어 판매되는 탕액에 대해서도 지속적인 모니터링이 필요할 것으로 판단된다.

• 한국식품영양학회지
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• 제33권2호
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• pp.215-221
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• 2020

The purpose of this study was to investigate the protective effect on oxidative stress induced PC12 cells, and volatile flavor composition of essential oils derived from medicinal plant seeds- Gossypium hirsutum L. (G. hirsutum), Coix lachryma-jobi (C. lachryma-jobi) and Oenothera biennis (O. biennis). The essential oils were obtained by the solvent (hexane) extraction method from the seeds. The essential oils of the seeds were analyzed by the solid-phase micro-extraction gas chromatography mass spectrometry (SPME-GC/MS). The major compounds of G. hirsutum, C. lachryma-jobi and O. biennis were cyclonexanol (16.65%), β-asarone (14.29%) and ylangene (50.01%). The DPPH radical scavenging activity (IC₅₀) was the highest value of 8.52 mg/mL in the O. biennis. Additionally, IC₅₀ values of G. hirsutum and C. lachryma-jobi were 26.76 mg/mL and 36.81 mg/mL. For the oxidative stress on PC12 cells, we treated with hydrogen peroxide (H₂O₂). The pretreatment of oxidative stress induced PC12 cells with all the essential oils preserved or increased their cell viability and G. hirsutum and O. biennis attenuated the ROS generation (by 68.75% and 56.25% vs. H₂O₂ control). The results of this study suggest that the essential oils derived from medicinal plant seeds could be used as valuable back data as a natural essential oil material to prevent neurodegenerative diseases by protecting neuro-cells.

• Radiation Oncology Journal
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• 제11권2호
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• pp.397-401
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• 1993

자궁경부암 환자의 병변의 범위 및 임상 경과를 추정하는데 TA-4의 유용성을 검토하고자 충남대학교병원에서 자궁경부암으로 진단 받은 58명의 환자를 대상으로 혈청 TA-4치를 RIA 방법으로 측정하여 $\le2\;ng/ml$을 정상 범위로 정하여 다음과 같은 결과를 얻었다. 1) 치료전 평평상피암 환자의 TA-4의 양성율은 $60\%,$ 선암은 $40\%$였다. 2) 병기가 진행될수록TA-4의 양성율및 평균치가 높아서, 양성율 병기 I는 $40\%,$ II는 $72\%,$ III는 $63\%,$ IV는 $100\%$였으며 평균치는 병기 I에서 3.1, II는 6.6, III는 8, IV는17.7 ng/ml였다. 3) 방사선 치료후 혈청내 TA-4치는 감소하여 5000 cGy 조사후 치료전 양성을 보였던 환자의 $44\%$에서 TA-4치 가 정상으로 돌아왔다. 4) 원발성 자궁경부암 환자의 양성율은 $59\%$였으나 지속성 또는 재발성등 치료에 실패한 15명 환자의 양성율은 $80\%$였다. 이상으로 연속적 혈청내 TA-4치의 측정은 자궁경부암 환자의 방사선치료에 따른 임상 경과를 관찰하는데 도움이 될 것으로 사료된다.

• Journal of Chest Surgery
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• 제33권8호
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• pp.648-654
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• 2000

Background: Cardiopulmonary bypass during open heart surgery causes systemic inflammatory respose. IL-10 is an anti-inflammatory cytokine that inhibits inflammatory process and protects organ function by down regulation of pro-inflammatory cytokine release and maintenance of blood level balance with pro-inflammatory cytokines. Mateial and Method: Plasma IL-10 levels were measured and analyzed in 22 patients who underwent open heart surgery (11 cases of coronary artery bypass graft, 11 cases of valve replacement) under cardiopulmonary bypass since 1988 January to July at Department of Thoracic and Czardiovascular surgery, Yeungnam University Hospital. 1g of methylprednisolone was administrated to thirteen patients randomly. Blood samp.es were taken and collected at the time of induction of anesthesia, 10 min before cardiopulmonary bypass, 10 min after starting of CPB, 10 min aftr aortic cross clamping, 10 min after ACC release, and 10 min, 2 hours, and `5 hours after CPB respectively. The plasma levels of IL-10 were determined by enzyme-linked immunosorbent assays(ELISA). Wilcoxon-Raule Sum test was used for statistical analysis. Result: In all 22 patients, cardiopulmonary bypass time was used for statistical analysis. Result: In all 22 patients, cardiopulmonary bypass time was 171$\pm$41.4 min and aortic cross clamp time was 118$\pm$36.5 min. Peak IL-10 level was achieved at 10 min after ACC(361.0$\pm$52.81pg/ml) and was decreased sharply at 2 hours after CPB. Peak IL-10 level was correlated positively with aortic cross clamp time(p=0.011); however, it did not correlated with bypass time(p=0.181). In valve replacement group, mean IL-10 level at peak point was 567.89$\pm$107.69 pg/ml and was significantly higher than that of coronary artery bypass group(205.67$\pm$192.70 pg/ml)(p<0.001). ACC time in valve replacement group was significantly longer than that of coronary artery bypass group(p<0.01), however, bypass time was not(p=0.212). Thirteen patients with steroid pretreatment before starting of CPB showed relatively higher plasma IL-10 level than in control group, however, no statistical significance was noted(p=0.19). Conclusion: plasma level of IL-10 was increased in association with cardiopulmonary bypass and revealed peak at 10 min after ACC release. IL-10 level was correlated positively with ACC time. Therefore, systemic inflammatory respeonse in association with cardiopulmonary bypass could be decreased by reducing ACC time during cardiac surgery.