• 한국식품과학회지
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• 제37권4호
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• pp.624-630
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• 2005

본 연구는 천연물에서의 생리활성물질 동정의 일환으로 Ficus Carica Linnoeus(FCL)로부터 60kDa의 FCL glycoprotein(무화과 당단백질)을 추출한 후, 무화과 당단백질의 첨가에 따른 과산화 지질 라디칼 억제능력 및 생질의 혈장 콜레스테롤 수준과 간 해독효소활성의 개선효과를 평가하였다. In vitro에서 리놀렌산 자동산화반응에 기초한 과산화 지질 라디칼 억제능력을 실험한 결과, 무화과 당단백질을 섭취시킨 농도가 증가함에 따라 과산화 지질 라디칼 억제율은 증가하였다. 한편, 생쥐에게 14일 동안 무화과 당단백질을 50 및 100mg/kg 농도로 섭취시킨 그룹과 무화과 당단백질을 섭취시킨 후 Triton WR-1339를 투여한 생쥐 그룹에서 혈액 및 간조직을 채취하여 혈장 콜레스테롤의 수준변화 및 해독효소의 활성을 측정한 결과, 100mg/kg 농도로 무화과 단백질을 섭취시킨 그룹에서 TC와 LDL-콜레스테롤의 수준은 유의적 감소효과가 나타났다(p<0.05). 또한 Triton WR-1339에 의해 고지혈증이 유발된 생쥐그룹에서도 TC와 LDL-콜레스테롤 수준이 유의적 억제능력을 보였는데, 특히 100mg/kg 농도에서 그 개선 효과는 더욱 분명하였다(p<0.01). 간의 해독효소 중 항산화 기능을 하는 SOD, CAT 그리고 GPx의 활성은 모두 증가되었는데, 특히 GPx는 100mg/kg의 농도에서 유의성을 보이며 증가하였다(p<0.01). 따라서 이러한 결과를 종합하면, 무화과 당단백질이 해득효소의 활성을 증가시킴으로써 체내의 ROS의 수준을 감소시키고, 이러한 항산화 효과가 혈중 콜레스테롤의 수준을 감소시키는데 영향을 미친 것으로 사료된다.

• 한국식품영양과학회지
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• 제31권3호
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• pp.527-533
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• 2002

전통차 제조용 식물 63종을 대상으로 macrophage lysosomal enzyme activity를 검색한 결과, 홍화 냉수 추출물에서 높은 macrophage lysosomal enzyme 활성 (219%)을 발견하여 냉수 추출물 CT-0 획분에 대하여 metahnol 환류, ethanol 침전, 투석, 동결건조를 실시하여 macrophage lysosomal enzyme 활성이 더욱 증가된(227%) 고분자 획분 CT-1을 얻었다. 이 획분의 macrophage활성화 성분의 본체를 파악하기 위하여 pronate처리에 의한 단백질 분해와 periodate를 이용한 당 부위의 선택적 실활 후 활성을 검토한 결과, pronase를 처리한 CT-1에서는 약 8% 정도의 활성 증가를 보인반면 periodate 산화물에서는 활성이 약 8% 정도 감소되는 것으로 보아 홍화로부터 macrophage 활성을 나타내는 냉수 추출물의 활성 본체는 다당임을 알 수 있었다. 조다당 활성획분에 대하여 anion exchange column chromatography를 실시하여 9개의 획분(CT-1-I~CT-1-VIII)을 얻었으며 수율과 활성이 가장 높은 CT-1-IIa 획분을 Sepharose CL-6B 및 Sephacrl S-200의 gel permeation chromatography를 수행하여 주요 활성 다당인 CT-1-IIa-2-1을 최종적으로 정제하였다. HPLC상에서 순수한 단일 peak로 확인된 CT-1-IIa-2-1은 분자량이 68 kDa정도의 다당인 것으로 나타났고 macrophage의 lysosomal enzyme 활성은 대조군을 100%로 비교했을 때 243%를 나타내었다. 또한 구성당의 조성은 xylose(27.4%), arabinose(16.1%), mannose(15.9%), glucose(14.5%)의 순이었다. 본 연구에서 CT-1-IIa-2-1은 macrophage의 면역활성을 증가시키는 물질임이 확인되었으나 mouse를 대상으로 급성독성 검사를 실시한 결과 LD$_{50}$값이 397mg/kg으로 일정 농도 이상의 고농도에서는 독성을 나타냈다. 따라서 본 시료에 대하여 아만성.만성 독성과 유전 및 면역 독성과 같은 구체적인 독성 검사를 실시하여 안전농도를 산출한다면, 면역증강물질로의 개발이 가능할 것으로 사료된다.다.

• Applied Microscopy
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• 제23권2호
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• pp.53-77
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• 1993

In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

• Clinical and Experimental Reproductive Medicine
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• 제32권3호
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• pp.207-216
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• 2005

Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries. Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in ${\alpha}$-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control. Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro. Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.

• 대한본초학회지
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• 제29권2호
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• pp.7-13
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• 2014

Objectives : Chronic mild stress (CMS) model is currently recognized as a better animal model of depression. The purpose of this study was to investigate the antidepressant-like effects of the Nelumbo nucifera leave extract using CMS model. Methods : The antidepressant-like effects of Nelumbo nucifera leaves extract was determined by using animal models of depression. Male ICR mice were divided into four groups: saline-treated normal, without CMS; saline-treated stress control; CMS+ Imipramine(20mg/kg); CMS+Nelumbo nucifera leaves extract(200mg/kg). All mice except the normal group exposed an unpredicted sequence of chronic mild stressors for 5 weeks. The behavior of mice were detected by sucrose preference test, forced swim test and tail suspension test. Then concentration of corticosterone in serum was detected by enzyme immunoassay. Results : Nelumbo nucifera leaves extract administration by daily gavage from the 3rd week exhibited an antidepressant-like effect on CMS-induced depression in mice. Nelumbo nucifera leaves extract administration at dose of 200 mg/kg significantly increased the sucrose consumption, and decreased the immobility durations in forced swim test and tail suspension test. Furthermore the corticosterone level decreased than control group. Conclusions : Chronic mild stress can affect mouse behavior and corticosterone level and cause depression. The present experiments not only further confirm the antidepressant-like effects of Nelumbo nucifera leaves extract in the sucrose preference test, forced swimming test and tail suspension test, but also the improving effects of Nelumbo nucifera leaves extract on the depression-like symptoms in the CMS model. Nelumbo nucifera leaves extract has the antagonism on CMS and produce antidepressive effects.

• 대한본초학회지
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• 제28권3호
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• pp.17-24
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• 2013

Objectives : This study was to evaluate the single dose toxicity of Persicae Semen (PS) in ICR mice. Methods : Aqueous extracts of PS (Yield = 18.60%) were administered as an oral dose of 2,000, 1,000 and 500 mg/kg (body weight) according to the recommendation of Korea Food and Drug Administration (KFDA) guidelines (2009-116, 2009). Animals were monitored for the mortality and changes in body weight, clinical signs and gross observation during 14 days after dosing, upon necropsy; organ weight and histopathology of 12 principle organs were examined. Results : Amygdalin contents in PS aqueous extracts were detected as $32.50{\pm}5.96{\mu}g/ml$. We could not find any PS extracts treatment related mortalities, clinical signs, changes on the body and organ weights, gross and histopathological observations up to 2,000 mg/kg in both female and male mice, except for transient and slight loss of locomotion detected in female and male mice treated with 2,000 mg/kg. In addition, pharmacological immunomodulatory effects related findings were also demonstrated in 2,000mg/kg treated female and male mice as hypertrophy and hyperplasia of lymphoid cells in the submandibular lymph nodes. Conclusions : Based on the results of this experiment, the approximate lethal dose (ALD) of PS extracts after single oral treatment in female and male mice were considered above 2,000 mg/kg, respectively. It should be carefully used in clinics because the possibilities of respiratory or neurological disorders were observed when administered over 2,000 mg/kg of PS extract related to amygdalin.

• 대한본초학회지
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• 제33권6호
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• pp.35-42
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• 2018

Objectives : The aim of this study was to investigate whether the extract of Coptidis Rhizoma inhibits inflammation in chronic cold stress (CCS)-exposed mice or not. Methods : Coptidis Rhizoma extract (CRE) was made by reflux with distilled water. Male ICR mice (7 weeks old) were divided randomly into 5 groups: (1) control, (2) CCS, (3) CCS+CRE 100 mg/kg, (4) CCS+CRE 300 mg/kg, (5) CCS+CRE 1,000 mg/kg groups. Mice were orally administered once a day for 14 days starting from 1 day before CCS. Group (2)-(5) were exposed to CCS conditions that maintained at $4^{\circ}C$ for 2 h once a day for 14 days. The levels of serum cortisol and hypothalamic prostaglandin E1 (PGE1) and PGE2 were measured by enzyme-linked immunosorbent assay kit. The expression levels of several pro-inflammatory factors like heat shock protein 70 (HSP70), c-fos, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured by western blot analysis in mouse hypothalamus. Results : Oral administration of CRE 1,000 mg/kg significantly suppressed the increase of serum cortisol levels in mice exposed to CCS. CCS-exposed mice had significantly increased the expression of HSP70, c-fos, and NF-kB in hypothalamus, while CRE treatment significantly attenuated the elevation of these pro-inflammatory factors. The ratio of PGE2/PGE1 was also higher in CCS-exposed mice than control group. CRE treatment significantly reduced the increase of PGE2/PGE1 ratio induced by CCS. Conclusion : These findings suggest that Coptidis Rhizoma may work as a potential agent to modulate inflammatory responses under the condition of cold adaptation formed by CCS.

• 한국환경보건학회지
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• 제49권1호
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• pp.22-29
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• 2023

Background: The existing research results on the combined toxicity of these pollutants using mammals, such as rodents, are insufficient, especially in relation to changes in the immune system. Objectives: This study aims at evaluating the cellular immune response to PE-MPs solely or when combined with Pb, which possess excellent adsorption capacity with PE-MPs and is commonly co-exposed in our daily lives. Methods: The study investigated the cellular immune function of 9-week ICR mice with 28 days exposure to PE-MPs (2 mg/mouse/day) and Pb (0.1 mM in distilled water) individually and in combination. PE-MPs were administered via gastric intubation while the lead intake was conducted via the oral drinking water route. Cellular immunity was evaluated by analyzing the production for T_H1 cytokines namely, TNF-α and IFN-𝛾 and T_H2 cytokines, IL-4 and IL-6 in culture supernatants from polyclonally activated splenic mononuclear cells ex vivo. Results: Both the PE-MPs only and the PE-MPs+Pb exposure group revealed an increased T_H1 response with elevated TNF-α and IFN-𝛾 levels and downregulated T_H2 response with low IL-4, and IL-6 production levels compared to the control group. Furthermore, an increased IFN-𝛾/IL-4 ratio was found in the PE-MPs only and PE-MPs+Pb exposure groups, which indicated the skewedness to T_H1 response. Meanwhile, reduced blood hemoglobin levels and increased levels of IL-4, the dominant T_H2 cytokine in the Pb-only exposure group, were observed. Conclusions: Our current findings on the predominance of T_H1 immune response in the PE-MPs and PE-MPs+Pb groups suggest that PE-MPs could be responsible for the predominant induction of the cellular immune changes. This finding could be used as an important landmark in research related to T_H1 predominance, such as autoimmune diseases. It suggests that additional research on immune modulation using longer exposure durations or the same exposure route is required to elucidate stronger findings.

• Parasites, Hosts and Diseases
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• 제62권1호
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• pp.42-52
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• 2024

Antimalarial drugs are an urgently need and crucial tool in the campaign against malaria, which can threaten public health. In this study, we examined the cytotoxicity of the 9 antimalarial compounds chemically synthesized using SKM13-2HCl. Except for SKM13-2HCl, the 5 newly synthesized compounds had a 50% cytotoxic concentration (CC₅₀) >100 μM, indicating that they would be less cytotoxic than SKM13-2HCl. Among the 5 compounds, only SAM13-2HCl outperformed SKM13-2HCl for antimalarial activity, showing a 3- and 1.3-fold greater selective index (SI) (CC₅₀/IC₅₀) than SKM13-2HCl in vitro against both chloroquine-sensitive (3D7) and chloroquine -resistant (K1) Plasmodium falciparum strains, respectively. Thus, the presence of morpholine amide may help to effectively suppress human-infectious P. falciparum parasites. However, the antimalarial activity of SAM13-2HCl was inferior to that of the SKM13-2HCl template compound in the P. berghei NK65-infected mouse model, possibly because SAM13-2HCl had a lower polarity and less efficient pharmacokinetics than SKM13-2HCl. SAM13-2HCl was more toxic in the rodent model. Consequently, SAM13-2HCl containing morpholine was selected from screening a combination of pharmacologically significant structures as being the most effective in vitro against human-infectious P. falciparum but was less efficient in vivo in a P. berghei-infected animal model when compared with SKM13-2HCl. Therefore, SAM13-2HCl containing morpholine could be considered a promising compound to treat chloroquine-resistant P. falciparum infections, although further optimization is crucial to maintain antimalarial activity while reducing toxicity in animals.

• 한국미생물·생명공학회지
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• 제44권3호
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• pp.236-245
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• 2016

밀배아유가 LPS로 자극된 대식세포에서 염증과 관련된 인자들에 미치는 영향을 살펴보기 위해, NO 및 전염증성 cytokine의 분비량과 iNOS 및 COX-2의 발현량, NF-κB p65와 MAPKs 활성화를 확인하였다. 또한 ICR 마우스를 이용하여 croton oil로 유도된 귀부종을 통해 귀두께 변화 및 귀조직을 관찰하였다. 그 결과, 밀배아유는 LPS로 인해 증가된 NO 및 전염증성 cytokine의 분비량을 감소시켰으며, 특히, IL-6와 TNF-α는 100 μg/ml 농도에서 90% 이상의 분비 억제효과를 나타내었다. LPS에 의해 증가된 iNOS와 COX-2의 발현은 100 μg/ml 농도로 밀배아유 처리시 발현 억제가 가장 크게 나타났으며, NF-κB p65의 발현도 밀배아유 농도가 증가함에 따라 발현 억제효과를 나타내었으며, 100 μg/ml 농도로 처리 시 억제 효과가 가장 컸다. 밀배아유는 MAPKs의 인산화를 억제시켰으며, 특히, ERK와 JNK의 인산화 억제가 농도 의존적으로 크게 효과를 나타내었다. 또한, 밀배아유는 croton oil로 유도된 귀부종에서 귀두께를 감소시켰으며, 경피와 진피의 두께 및 진피에 침윤된 비만세포의 수도 감소시킴을 확인하였다. 이상으로, 밀배아유는 NF-κB 및 MAPKs의 신호 경로 조절을 통한 염증성 매개 인자들의 발현 감소로 항염증 효과를 나타냄을 확인하여, 천연 항염증 소재로서 활용도가 가능함을 시사한다.