• Journal of Plant Biology
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• 제36권3호
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• pp.233-239
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• 1993

Effect of Ca2+ on the conversion of 1-aminocycloprophane-1-carboxylic acid (ACC) to ethylene was studied with 2.5 day-old mung bean hypocotyl segments. The conversion of ACC in these tissues was inhibited by plasmolysis and sulfosuccinimidyl (hydroxyphenyl) propionate (sulfo-SHPP). The ACC induced ethylene production in HC (high calcium)-tissue grown on the Ca2+ added medium was greater than that in N (normal)-tissue. HC-tissue had a lower inhibition rate of ACC conversion by EGTA and Ca2+ -channel blockers than N-tissue. The rates of the ACC conversion by both kinds of tissues were stimulated by the Ca2+ ionophore A23187. From these results, we suggests a mechanism for the stimulative effect of Ca2+ on the conversion of ACC to ethylene as follows; in some tissues where ACC conversion is linked with plasma membrane, Ca2+ may be transported from apoplast through Ca2+ -channel into the cytoplasm ad stimulate ACC-oxidase activity.

• Archives of Pharmacal Research
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• 제17권6호
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• pp.405-410
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• 1994

The present study was undetertaken to demonstrate whether or not angiotensin II activates a phopholipase D in rabbit kidney proximal tubule cells. By measuring the formation of [$^3H$] phosphatidic acid and [$^3H$]diacylglycerol. This result suggests that some phosphatidic acid seems to be formed directly from phosphatidylcholin by the action of phopholipase D, not from the action of diacylglycerol kinase on the diacylglycerol. In addition the other mechanisms by which phospholipase D is activated was examined. We have found that phospholipase D was activited by extracellular calium ion. It has also been shown that angiotensin II may activate phosphoilpase D through protein kinase C-independent pathway.

• The Korean Journal of Physiology and Pharmacology
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• 제20권2호
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• pp.213-220
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• 2016

Mast cells are primary mediators of allergic inflammation. Beta-1,3-glucan (BG) protects against infection and shock by activating immune cells. Activation of the BG receptor induces an increase in intracellular $Ca^{2+}$, which may induce exocytosis. However, little is known about the precise mechanisms underlying BG activation of immune cells and the possible role of mitochondria in this process. The present study examined whether BG induced mast cell degranulation, and evaluated the role of calcium transients during mast cell activation. Our investigation focused on the role of the mitochondrial calcium uniporter (MCU) in BG-induced degranulation. Black mouse (C57) bone marrow-derived mast cells were stimulated with $0.5{\mu}g/ml$ BG, $100{\mu}g/ml$ peptidoglycan (PGN), or $10{\mu}M$ A23187 (calcium ionophore), and dynamic changes in cytosolic and mitochondrial calcium and membrane potential were monitored. BG-induced mast cell degranulation occurred in a time-dependent manner, and was significantly reduced under calcium-free conditions. Ruthenium red, a mitochondrial $Ca^{2+}$ uniporter blocker, significantly reduced mast cell degranulation induced by BG, PGN, and A23187. These results suggest that the mitochondrial $Ca^{2+}$ uniporter has an important regulatory role in BG-induced mast cell degranulation.

• Journal of Ginseng Research
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• 제22권2호
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• pp.133-139
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• 1998

We have studied an activation mechanism of $pp60^{c-src}$ protein tyroslne kinase (PTK) by ginsenoside-$Rg_1$ (G-$Rg_1$ ) in NIH(pMcsrc/foc)B c-src overexpressor cells. It was previously reported that G--$Rg_1$ stimulated the activation of c-src kinase at 20 pM with a 18 hr-incubation, increasing the activity by 2-4-fold over that of untreated control, and this effect was blocked by treatments of in- hibitors of either protein synthesis (cycloheximide) or RNA synthesis (actinomycin D) (Hong, H.Y. et at. Arch. Pharm. Res. 16, 114 (1993)). However, an amount of c-src protein itself in wild-type cells was not changed by G-$Rg_1$. When the cells mutated at one or two tyrosine residue(s) (Y416/527) that are important sites to regulate the kinase activity were treated with G-$Rg_1$, increases both in the activity of c-src kinase and in the expression of the protein were not observed. In addition, removal of extracellular calcium ion by EGTA or inhibition of PKC by H-7 canceled the G-$Rg_1$-induced activation of the kinase. Although the activation was little affected by G-$Rg_1$ with a calcium ionophore A23187, it was synergistically stimulated by treatment of G-Rgl and PMA, a PKC activator. Taken together, these results suggest that the activation of c-src kinase by G-$Rg_1$ is caused by an increase in the specific activity of the kinase, but not in amount of it, and is involved with both collular calcium ion and PKC. Further the increase in the specific activity of c-src kinase may result from altered phosphorylation at tyro-416 and -527.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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• pp.251-260
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• 1998

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol $(E_2)$ were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with $10{\mu}l$ of 10% DMSO was induced spontaneously in $10.1{\pm}9.8%$, and acrosomal reaction with calcium ionophore A 23187 was induced in $27.4{\pm}18.1%$, and the ARIC value was $17.4{\pm}16.2%$. There were no significant correlation between the ARIC value and the fertilization rate ($r^2$=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos ($r^2$=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the micro assisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.

• 대한약리학회지
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• 제29권2호
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• pp.165-174
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• 1993

칼슘이온은 신경섬유 성장의 중요한 조절인자이나 그 정확한 작용기전은 불명확하다. 세포골격 단백은 in vivo 및 in vitro에서 칼슘의존성 단백분해효소(칼파인)에 의해 신속히 분해되므로, 칼슘이온에 의한 신경섬유의 퇴행에 있어서 칼파인의 관련성을 추구하기위하여, 배양된 해마신경세포에서 칼슘이온 ionophore인 A23187에 의한 신경섬유의 성장억제가 칼파인의 억제제인 EST 및 MDL 28170에 의해 차단되는지를 조사하였다. A23187은 100nM의 농도에서 축삭에는 영향이 없이 수상돌기의 퇴행을 유발하였으나, 300 nM의 농도에서는 축삭의 성장을 억제하였다. EST(5 혹은 20 uM) 및 MDL 28170(20 uM)은 100 nM A23187의 수상돌기에 대한 작용과 300 nM A23187의 축삭에 대한 작용을 효과적으로 차단하였다. EST는 A23187에의한 세포내 칼슘이온의 증가를 차단하지 못하였다. 이상의 결과는 해마추상체세포에서 칼슘에 의한 신경섬유의 퇴행이 칼파인에 의해 매개됨을 시사한다.

• 한국가축번식학회지
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• 제27권1호
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• pp.25-34
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• 2003

인체 트롬보포이에틴(hTPO)은 megakaryopoiesis 과정에 주요한 역할을 하는 사이토카인이다. 따라서 이러한 트롬보포이에틴을 유선조직에서 직접적으로 발현시키기 위하여 소 베타 카제인 프로모터, 인체 트롬보포이에틴 cDNA 및 네오유전자로 구성된 발현벡터를 구축하였다. 소 귀조직 세포로부터 유도된 섬유아세포에 lipoffctamine을 이용하여 발현벡터(pBT-L n대)의 삽입을 유도하였다. G4l8 저항성을 지닌 세포의 콜로니 형성을 유도하기 위하여 2주 이상 배양을 실시하였다. 형질전환 콜로니는 PCR에 의해 동정하였으며, 이들 콜로니를 핵치환 전까지 계속적으로 증식을 유도하였다. 형질전환 세포에 의해 재구성된 난자는 전기적인 융합과 calcium ionophore와 6-DMAP를 이용한 활성화를 실시하였으며, 체외에서 7일간 배양을 실시하였다. 총 35개의 콜로니를 PCR에 의해 분석한 결과, 이 중 29(82.9%)개가 형질전환된 콜로니였다. 형질전환된 세포로 재구성된 난자의 난할율 및 배반포로의 발달율은 65.1%와 23.8%로 나타났다. 형질전환된 세포로 재구성된 난자로부터 발달한 29개의 배반포 중 27개가 형질전환으로 확인되었다. 따라서 이러한 결과들은 형질전환 소 수정란을 형질전환된 세포를 이용한 체세포 복제 기법을 통해 효과적으로 생산할 수 있다는 것을 제시하고있다.

• Clinical and Experimental Reproductive Medicine
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• 제21권1호
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• pp.1-11
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• 1994

The occurrence and time course of capacitation, acrosomal loss, and hyperactivated motility require quantitative definition in order to characterize fertile human sperm. Recently the method has been developed to estimate the quality of spermatozoa by using kinematic parameters such as curvilinear velocity(VCL), average path velocity(VAP), linearity(LIN), straightness(STR), amplitude of lateral head displacement(ALH), and beat cross frequence(BCF) from Computer Assisted Sperm Analysis (CASA). In this study, using the Hamilton Thorn motility analyzer HTM 2030(Hamilton Thorn Research, Beverly, MA), we attempted to identify the spermatozoa with hyperactivated motility (HA) objectively and to monitor hyperactivation of human spermatozoa during incubation in capacitating media and after treatment of calcium ionophore as compared with acrosome status. And we examined whether HA are related to the result of SPA. Semen samples obtained from 16 healthy men were prepared by swim up technique and preincubated in a capacitating media(modified BWW medium) for 5 hours and treated with calcium ionophore solution. The acrosome reaction was detected with PSA-FITC labelling of the acrosome and in vitro sperm ferilizing capacity was assessed by the zona free hamster ovum penetration assay (SPA). The incidence of hyperactivated sperm was 2.6% in fresh semen, 14.3% of the swim up population, 13.7% after 5h of incubation. Significant increase of percentage of hyperactivated sperm was observed after the incubation (p<0.05) but after treatment, no significant changes of percentage of hyperactivated sperm(l1.8%) in contrast to significant rise in the percentage of acrosome reacted cells. Correlation analysis failed to show any significant relationship between the percentage of sperm with HA and SPA score. In conclusion, although no direct correlations were found between the results of SPA and HA, hyperactivation of sperm is associated with capacitation and monitoring hyperactivated sperm will be expected as a method of evaluating the functional quality of sperm such as SPA.

• 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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• 제48권6호
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• pp.590-596
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• 2016

본 연구에서는 우리나라에서 가장 많이 재배되고 있는 캠벨어얼리 포도 품종에서 포도가지를 대상으로 산화방지, 항염증 및 항가려움증 효과를 검증하고 활성물질을 나타내는 지표물질을 추적 조사하였다. 그 결과 포도가지 추출물의 총 폴리페놀과 총 플라보노이드 함량은 각각 $201.42{\pm}4.16$과 $11.85{\pm}0.44mg\;GAE/g$으로 조사되었다. 또한 캠벨어얼리 포도가지 추출물의 ABTS와 DPPH 라디칼 소거 활성은 각각 $45.60{\pm}0.09$ ($IC_{50}$)과 $299.13{\pm}0.22$ ($IC_{50}$)으로 나타나 산화방지 활성이 우수하였다. 게다가 캠벨어얼리 포도가지 추출물은 지방질다당류로 활성화된 RAW 264.7 세포에서 전염증성 매개물인 산화질소와 프로스타글란딘 $E_2$를 iNOS와 COX-2 분자 발현 억제를 통하여 억제하였고, 전염증성 사이토카인인 인터류킨-1베타와 인터류킨-6를 농도 의존적으로 억제하는 효능이 있었다. 더욱이 phorbol 12-myristate 13-acetate (PMA)와 calcium ionophore A23187로 활성화된 인간 유래 비만세포인 HMC-1 세포에서 종양괴사인자-알파와 인터류킨-6를 농도 의존적으로 억제하는 효능이 있었다. 마지막으로 Compound 48/80으로 유도되는 마우스 가려움증을 캠벨어얼리 포도가지 추출물은 효과적으로 억제하였다. 이러한 캠벨어얼리 추출물에서 활성을 나타내는 물질을 추적한 결과 레스베라트롤의 함량이 높게 검출되었다. 그러므로 본 연구의 결과를 종합해 볼 때 캠벨어얼리 포도가지 추출물은 아토피 질환에서 나타나는 염증과 가려움증을 효과적으로 제어할 수 있는 효과적인 소재임을 제시하였다.