• 한국응용곤충학회지
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• 제55권4호
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• pp.413-419
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• 2016

Chlorantraniliprole은 곤충 근육의 $Ca^{2+}$ 농도를 조절하는 Ryanodine 수용기(RyR)에 작용 하는 diamide계통의 작물보호제이다. 최근에 보고된 chlorantraniliprole 저항성 배추좀나방 계통은 RyR에 돌연변이를 가지고 있다. 본 연구에서는 초파리를 모델 곤충으로 저농도와 고농도의 chlorantraniliprole로 도태된 두 종류의 저항성 계통을 확보하였다. 두 종류의 저항성 계통은 접촉독성과 섭식독성 평가법을 활용하여 저항성 지수를 산출하였다. 접촉 독성 평가에서 두 종류의 저항성 계통은 대조군과 비교하여 95% 신뢰구간에서 저항성 발달에 차이가 없었지만, 섭식 독성 평가의 경우에서는 고농도 저항성 계통과 저농도 저항성 계통에서 대조군 대비 각각 2.1배와 8.1배의 통계적으로 유의한 저항성 증가가 나타났다. 작용점 유전자인 RyR 발현량 비교 결과, 두 종류의 저항성 계통에서 RyR의 발현량이 유의하게 감소하였고, 주요 약제 관련 효소인 Acetylcholinesterase와 Glutathione-S-transferase 활성은 조직 특이적으로 증가하는 것을 확인하였다. 이러한 결과들은 초파리에서 chlorantraniliprole에 대한 섭식독성 저항성의 발달에는 주요 해독 관련 효소의 과활성도 관여할 것 임을 보여주고 있다.

• The Korean Journal of Physiology
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• 제27권2호
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• pp.175-183
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• 1993

There are many report suggesting that influx and intracellular calcium concentration $([Ca^{2+}]_i)$ are related to cell signalling in various cells. However, it has not been reported that calcium channel activation is affected by the substances involved in signal transduction pathways in the mouse eggs. In this study, the effects of isoprenaline (ISP) and cyclic AMP on calcium influx through calcium channels were investigated to show their relationship with the signal transduction process in unfertilized mouse eggs. Using whole cell voltage clamp techniques, calcium currents, elicited by the depolarizing pulses of 300 ms duration (from -50 mV to 50 mV in 10 mV increments) from a holding potential of -80 mV, were recorded. The current-voltage (I-V) relation of calcium currents was shown to be bell-shaped; the current began to activate at -50 mV and reached its maximum $(-1.33{\pm}0.16\;nA:\;mean{\pm}S.E.,\;n=7)$ at -10 mV, then decayed at around 50 mV. Calcium currents were fully activated within $7\;ms{\sim}20\;ms$ and completely inactivated 200 ms after onset of the step pulse. ISP within the concentration ranges of $10^{-8}\;M{\sim}10^{-4}\;M$ dose-dependently increased the amplitude calcium current. The permeable cyclic AMP analogue,8-bromocyclic AMP, also increased its maximal amplitude by 46ft at $10^{-5}\;M$, while protein kinase inhibitor (PKI), which is known to inhibit 0.02 phosphorylating units of cyclic AMP-dependent protein kinase (PKA) per microgram decreased calcium currents. Currents recorded in the presence of PKI were resistant to increase by the application of $10^{-5}\;M$. Also, PKI inhibited the calcium current increase elicited by ISP treatment. These results suggest that $\beta-adrenergic$ regulation of the calcium channel is mediated by the cAMP-dependent protein kinase. This signal transduction pathway might play a role in regulating $[Ca^{2+}]_i$, level due to the increase of calcium influx in mouse eggs.

• The Korean Journal of Physiology and Pharmacology
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• 제1권6호
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• pp.825-830
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• 1997

Leukotrienes(LTs) are hewn to act as a mediator provoking tissue response in inflammation. This finding implicates that LTs also play important roles in the pathogenesis of H, pylori-induced gastritis and gastric ulceration. Rebamipide is being currently used as a therapeutics for gastritis and peptic ulcer, but their mechanisms of action have not been known clearly yet. One possibility is that their therapeutic effects are ascribed to interfering with the H. pylori-induced release of LTs from neutrophils and gastric mucosal cells. In the present study, this possibility was tested using $LTB_4$ as the test material in human neutrophils and Kato III cells(gastric adenoma cells as a substitute for gastric mucosal cells). The release of $LTB_4$ from both neutrophils and Kato III cells was time and H. pylori-dose dependent. The maximum release of $LTB_4$ was induced by neutrophils and Kato III cells when these cells incubated with H. pylori $(4.8{\times}10^8\;cells/ml$ for 30min. But in the presence of rebamipide the release of $LTB_4$ from these cells was suppressed in dose dependent manners. The release was completely suppressed at 1.0 mM of rebamipide in neutrophils and 2.0 mM of this drug in Kato III cells, respectively. We also obtained the results that the release of $LTB_4$ was induced by A23187$(Ca^{2+}\;ionophore)$ and the A23187-induced release was also inhibited by rebamipide. It seems that the machanism of action of rebamipide is through its interaction with the level of intracellular $Ca^{2+}$. In view of the roles of $LTB_4$ in inflammatory reaction and the roles of H. pylori in gastritis and peptic ulcer, the effects of this drug observed in this study may contribute to their therapeutic action in these gastric disorders.

• The Korean Journal of Pain
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• 제29권4호
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• pp.229-238
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• 2016

Background: The goal of this in vitro study was to investigate the effect of lipid emulsion on vasodilation caused by toxic doses of bupivacaine and mepivacaine during contraction induced by a protein kinase C (PKC) activator, phorbol 12,13-dibutyrate (PDBu), in an isolated endothelium-denuded rat aorta. Methods: The effects of lipid emulsion on the dose-response curves induced by bupivacaine or mepivacaine in an isolated aorta precontracted with PDBu were assessed. In addition, the effects of bupivacaine on the increased intracellular calcium concentration ($[Ca^{2+}]_i$) and contraction induced by PDBu were investigated using fura-2 loaded aortic strips. Further, the effects of bupivacaine, the PKC inhibitor GF109203X and lipid emulsion, alone or in combination, on PDBu-induced PKC and phosphorylation-dependent inhibitory protein of myosin phosphatase (CPI-17) phosphorylation in rat aortic vascular smooth muscle cells (VSMCs) was examined by western blotting. Results: Lipid emulsion attenuated the vasodilation induced by bupivacaine, whereas it had no effect on that induced by mepivacaine. Lipid emulsion had no effect on PDBu-induced contraction. The magnitude of bupivacaine-induced vasodilation was higher than that of the bupivacaine-induced decrease in $[Ca^{2+}]_i$. PDBu promoted PKC and CPI-17 phosphorylation in aortic VSMCs. Bupivacaine and GF109203X attenuated PDBu-induced PKC and CPI-17 phosphorylation, whereas lipid emulsion attenuated bupivacaine-mediated inhibition of PDBu-induced PKC and CPI-17 phosphorylation. Conclusions: These results suggest that lipid emulsion attenuates the vasodilation induced by a toxic dose of bupivacaine via inhibition of bupivacaine-induced PKC and CPI-17 dephosphorylation. This lipid emulsion-mediated inhibition of vasodilation may be partly associated with the lipid solubility of local anesthetics.

• 대한한방종양학회지
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• 제4권1호
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• pp.177-197
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• 1998

This experimental study was carried out to evaluate the effect of Yipahnsan on angiogenic inhibition mechanism. This study investigates the effects of Yipahnsan on angiogenic inhibition mechanism evaluate cell adhesive inhibition effect, DNA fragmantaion analysis, nuclear condensation assay, FACScan analysis, angiogenic lumen formation assay, immunocytochemistry analysis, RT-PCR for mRNA expression, western blot analysis, confocal analysis for $Ca^{2+}influx$. The results were summarized as follows : 1. The cell adhesive inhibition ability was strongly increased from $5{\mu}g/ml$ on ECV304 cell line and ECVPAR cell line. 2. YY water extract caused $G_0/G_1$ arrest peak to existed on the ECV304 cell line. 3. YY water extract caused inhibition of proliferation and inducement of apoptosis on the collagen coated plate in ECV304 cell line. 4. YY water extract inhibited the lumen formation on the matrigel coated plate in ECV304 cell line. 5. YY water extract inhibited the expressions of LFA-1 and ELAM-1 on ECV304 cell line and ECVPAR cell line. 6. YY water extract inhibited the expressions of MMP-9 and uPA on ECV304 cell line and ECVPAR cell line. 7. YY water extract inhibited the expression of integrin ${\alpha}_v{\beta}_3$ on ECV304 cell line and ECVPAR cell line. 8. YY water extract decreased the change of $Ca^{2+}$ in intracellular on ECV304 cell line and ECVPAR cell line. According to the results, Yipahnsan showed to be a key antagonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA were blocked under the angiogenesis model. Thus, we suggested that Yipahnsan blocks angiogenesis by inducing apoptosis in ECV304 and ECVPAR cell lines, and another oriental herbal medicine that treats qi-stagnation and blood-stasis type also has angiogenic inhibition effects.

• 대한한방종양학회지
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• 제4권1호
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• pp.17-36
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• 1998

This experimental study was carried out to evaluate the effects of Hwallakhyoreungdan on angiogenic inhibition mechanism. In order to investigate the effects of Hwallakhyoreungdan on angiogenic inhibition mechanism, MTT assay, cell adhesive inhibition effect, DNA fragmantaion analysis, Nuclear condensation assay, FACScan analysis, Angiogenic lumen formation assay, Immunocytochemistry analysis, RT-PCR for mRNA expression, Western blot analysis and Confocal analysis for $Ca^{2+}$ change were performed. The results were summarized as follows: 1. The cell adhesive inhibition ability was strong from $5{\mu}g/ml$. 2. The $G_0/G_1$ arrest peak was existed on ECV304 cell-line. 3. The cells on Collagen plate were inhibition of proliferation and inducement of apoptosis by HR water extract. 4. Angiogenic lumen formation was inhibited by HR water extract. 5. LFA-1 and ELAM-1's expression were inhibited by HR water extract. They are commenly participation on inflammation and tumor regeneration. 6. The expression of MMP-9 and uPA were inhibited by HR water extract. 7. The expression of integrin ${\alpha}_v{\beta}_3$ was inhibited by HR water extract. 8. The expression of intracellular molecule were successively inhibited by HR water extract therefore the proliferation of ECV304 cell line was stopped and apoptosis was induced. 9. The change of $Ca^{2+}$ was decreased by HR water extract it cause confusion of signal transduction pathway therefore it was take part in apoptosis. According to the results, Hwallakhyoreungdan showed to be a key antaonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA was blocked under the angiogenesis model. Thus, we suggests that Hwallakhyoreungdan blocks angiogenesis by inducing apoptosis of ECV304 and ECVPAR cell lines and another oriental herbal medicine that treats blood-stasis type also has angiogenic inhibition effects.

• The Korean Journal of Pain
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• 제27권3호
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• pp.229-238
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• 2014

Background: A toxic dose of bupivacaine produces vasodilation in isolated aortas. The goal of this in vitro study was to investigate the cellular mechanism associated with bupivacaine-induced vasodilation in isolated endothelium-denuded rat aortas precontracted with phenylephrine. Methods: Isolated endothelium-denuded rat aortas were suspended for isometric tension recordings. The effects of nifedipine, verapamil, iberiotoxin, 4-aminopyridine, barium chloride, and glibenclamide on bupivacaine concentration-response curves were assessed in endothelium-denuded aortas precontracted with phenylephrine. The effect of phenylephrine and KCl used for precontraction on bupivacaine-induced concentration-response curves was assessed. The effects of verapamil on phenylephrine concentration-response curves were assessed. The effects of bupivacaine on the intracellular calcium concentration ($[Ca^{2+}]_i$) and tension in aortas precontracted with phenylephrine were measured simultaneously with the acetoxymethyl ester of a fura-2-loaded aortic strip. Results: Pretreatment with potassium channel inhibitors had no effect on bupivacaine-induced relaxation in the endothelium-denuded aortas precontracted with phenylephrine, whereas verapamil or nifedipine attenuated bupivacaine-induced relaxation. The magnitude of the bupivacaine-induced relaxation was enhanced in the 100mM KCl-induced precontracted aortas compared with the phenylephrine-induced precontracted aortas. Verapamil attenuated the phenylephrine-induced contraction. The magnitude of the bupivacaine-induced relaxation was higher than that of the bupivacaine-induced $[Ca^{2+}]_i$ decrease in the aortas precontracted with phenylephrine. Conclusions: Taken together, these results suggest that toxic-dose bupivacaine-induced vasodilation appears to be mediated by decreased calcium sensitization in endothelium-denuded aortas precontracted with phenylephrine. In addition, potassium channel inhibitors had no effect on bupivacaine-induced relaxation. Toxic-dose bupivacaine-induced vasodilation may be partially associated with the inhibitory effect of voltage-operated calcium channels.

• 대한화장품학회지
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• 제44권2호
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• pp.191-200
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• 2018

본 연구에서는 제1형 프로콜라겐 분비량 측정을 통해 피부 주름개선 효과가 있는 천연물을 검색하던 중, 연꽃잎추출물(Nelumbo nucifera leaves extract, NLE)이 가장 우수한 효능을 나타냄을 확인하였다. 고압액체크로마토그래피로 NLE를 분석한 결과 isoquercitrin이 함유되어 있었으며, isoquercitrin도 normal human dermal fibroblast (NHDF)에서 제1형 프로콜라겐의 분비를 촉진한다는 것을 확인할 수 있었다. 또한 NLE와 isoquercitrin은 자유 라디컬 소거 활성을 가지고 있었으며, 특히 isoquercitrin은 인간 피부유래 세포주인 HaCaT 및 NHDF에서 활성산소종의 생성을 억제하고 총항산화능을 증가시켰다. 최종적으로 본 연구팀은 isoquercitrin의 함유량을 높이기 위해 정제한 NLE를 1.7% 함유한 크림(isoquercitrin 51 ppm 함유)을 제조하여 눈가의 주름개선 효과를 평가하였다. 30 ~ 65세의 한국인 여성 22명을 대상으로 얼굴의 한쪽 면엔 NLE 함유 크림을, 그리고 다른 한쪽 면엔 대조제품을 매일 2회씩, 8주간 연속 사용하도록 하였다. 그 결과 NLE 함유크림을 사용하였을 때 피부자극이 발견되지 않았으며, 대조제품과 비교하여 통계적 유의성 있게 눈가의 주름을 감소시키는 것을 확인할 수 있었다. 본 연구결과는 NLE 및 isoquercitrin이 우수한 콜라겐 생성 촉진 및 항산화 효능을 가지며, isoquercitrin 함유 NLE는 안전한 천연 주름개선 화장품 소재로 적용 가능하다는 것을 시사한다.

• 한국식품과학회지
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• 제32권1호
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• pp.218-224
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• 2000

분자량이 상이한 세 종류의 키토산의 항균활성을 E. coli O157 : H7, S. aureus 및 C. albicans 균주를 이용하여 측정, 분석하였다. E. coli O157 : H7와 S. aureus에 대해서는 분자량 10,000인 키토산이 가장 강한 항균활성을 보였으며 C. albicans에 대해서는 6량체의 키토산 올리고당이 가장 강한 활성을 나타내었다. 키토산 첨가농도는 E. coil O157 : H7와 S. aureus의 경우 0.1 mg/mL의 농도에서, C. albicans의 경우는 chitohexaose 1 mg/mL의 농도에서 항균활성이 가장 높았다. 모든 키토산 처리구에서 미생물 사멸 속도는 키토산 처리 1시간 이내에서 가장 높게 나타났으며 그 이후로는 점차 낮은 속도를 보였다. 사멸되었거나 파손된 미생물 세포로부터 유래되는 단백질, 핵산물질 및 $Ca^{2+}$ 량은 키토산 처리 시간 이내에 가장 많았으며 ${\beta}-galactosidase$ 활성도 같은 시간대에서 가장 빠른 속도로 중대되는 것으로 나타났다. S. aureus에 대한 세포막 손상 정도를 측정하여 본 결과 전체 미생물균체중 약 10%에 상당하는 균수가 막손상을 가져 왔다. 따라서, 키토산은 고유의 양이온성 성질을 이용하여 미생물의 세포벽과 세포막에 결합하여 그 결과로 세포 내 물질의 세포 외로의 유출내지는 세포막 대사의 저해 등의 효과를 나타냄으로써 항균활성을 나타내는 것으로 추측되었다.

• Journal of Ginseng Research
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• 제44권3호
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• pp.490-495
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• 2020

Background: Ginsenoside Rk1, a saponin component isolated from heat-processed Panax ginseng Meyer, has been implicated in the regulation of antitumor and anti-inflammatory activities. Although our previous studies have demonstrated that ginsenoside Rg3 significantly attenuated the activation of NMDA receptors (NMDARs) in hippocampal neurons, the effects of ginsenosides Rg5 and Rk1, which are derived from heat-mediated dehydration of ginsenoside Rg3, on neuronal NMDARs have not yet been elucidated. Methods: We examined the regulation of NMDARs by ginsenosides Rg5 and Rk1 in cultured rat hippocampal neurons using fura-2-based calcium imaging and whole-cell patch-clamp recordings. Results: The results from our investigation showed that ginsenosides Rg3 and Rg5 inhibited NMDARs with similar potencies. However, ginsenoside Rk1 inhibited NMDARs most effectively among the five compounds (Rg3, Rg5, Rk1, Rg5/Rk1 mixture, and protopanaxadiol) tested in cultured hippocampal neurons. Its inhibition is independent of the NMDA- and glycine-binding sites, and its action seems to involve in an interaction with the polyamine-binding site of the NMDAR channel complex. Conclusion: Taken together, our results suggest that ginsenoside Rk1 might be a novel component contributable to the development of ginseng-based therapeutic treatments for neurodegenerative diseases.