• The Korean Journal of Physiology and Pharmacology
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• 제20권2호
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• pp.185-192
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• 2016

Ampicillin, a ${\beta}$-lactam antibiotic, dose-dependently protects neurons against ischemic brain injury. The present study was performed to investigate the neuroprotective mechanism of ampicillin in a mouse model of transient global forebrain ischemia. Male C57BL/6 mice were anesthetized with halothane and subjected to bilateral common carotid artery occlusion for 40 min. Before transient forebrain ischemia, ampicillin (200 mg/kg, intraperitoneally [i.p.]) or penicillin G (6,000 U/kg or 20,000 U/kg, i.p.) was administered daily for 5 days. The pretreatment with ampicillin but not with penicillin G significantly attenuated neuronal damage in the hippocampal CA1 subfield. Mechanistically, the increased activity of matrix metalloproteinases (MMPs) following forebrain ischemia was also attenuated by ampicillin treatment. In addition, the ampicillin treatment reversed increased immunoreactivities to glial fibrillary acidic protein and isolectin B4, markers of astrocytes and microglia, respectively. Furthermore, the ampicillin treatment significantly increased the level of glutamate transporter-1, and dihydrokainic acid (DHK, 10 mg/kg, i.p.), an inhibitor of glutamate transporter-1 (GLT-1), reversed the neuroprotective effect of ampicillin. Taken together, these data indicate that ampicillin provides neuroprotection against ischemia-reperfusion brain injury, possibly through inducing the GLT-1 protein and inhibiting the activity of MMP in the mouse hippocampus.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.41-49
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• 1996

본 연구는 체외수정에 의해 생산된 생쥐 배반포기배를 vitrification 방법으로 동결보존하였을때 높은 생존율을 얻기 위한 적정조건을 검토하고자 실시하였다. 배반포기배를 생산하기 위하여, B6CBA F1 (C57BL/6, (표현불가)${\times}CBA/N$, (표현불가)) 계통의 생쥐 미수정란에 $1{\times}10^6$ spermatozoa/ml 농도의 정자로서 수정을 유도하였으며, 이후 $37^{\circ}C$, 5% $CO_2$배양기내에서 96시간동안 체외배양하였다. 배양 4일째의 배반포기배는 발달상태에 따라 early, middle 그리고 hatching blastocysts로 구분하였다. 본 실험에 사용된 동결보존액은 30% Ficoll과 0.5mol의 sucrose가 첨가된 mDPBS 용액에 40%의 ethylene glycol를 첨가한 EFS 40 (Zhu et al., 1993) 이었고, 수정란은 $25^{\circ}C$의 상온에서 먼저 20% ethylene glycol에 노출된 후 EFS 40 용액으로 옮겨 액체질소에 침지하는 2단계 동결법에 의해 동결보존되었으며, 급속융해하여 다음과 같은 결과를 얻었다. 1. 체외수정율과 배양 4일째 배반포기까지의 배발달율은 각각 89.4%와 86.1%였다. 2. 20% ethylene glycol에서 5분간 평형된 후 EFS 40 용액에 냉동보존된 후 융해된 난자의 생존율은 20% ethylene glycol에 0, 1, 3분간 평형된 난자의 생존율에 비해 유의하게 높았다. 3. 배반포기배를 20% ethylene glycol에서 5분간, EFS 40 용액에 1분간 차례로 노출한 다음 체와배양하였던 바, 배양 24시간째 생존율은 $82.9%{\sim}88.4%$ 였다. 본 연구 결과, 체외수정, 배양된 생쥐 배반포기배는 20% ethylene glycol과 EFS40에 대한 노출만으로는 난자의 생존성에 나쁜 영향을 미치지 않는 것으로 미루어 보아 배반포기배의 초자화 동결이 가능함을 시사하였다. 따라서 동결 융해 후 높은 생존율은 상온에서 난자를 2단계 즉, 20% ethylene glycol에 5분간 평형시킨 후 EFS 40 용액에 노출하여 1분내에 LN2에 직접 침지하는 간편한 동결방법으로 얻을 수 있었다.

• 동의생리병리학회지
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• 제22권2호
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• pp.350-356
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• 2008

Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Cheonghyul-san has been known to be effective for the antidiabetic, antihyperlipidemic and antiobesitic prescription, and composed of four crude herbs. In male ob/ob mouse with severe obesity, hyperinsulinemia, hypergiycemia, hyperlipidemia, the acting mechanisms of Cheonghyul-san were examined. Mice were grouped and treated for 5 weeks as follows. Both the lean (C57/BL6J black mice) and diabetic (ob/ob mice) control groups received standard chow. The experimental groups were fed with a diet of chow supplemented with 7.5, 15 and 30 mg Cheonghyul-san per 1 kg of body weight for 14 days. The effects of Cheonghyul-san extract on the ob/ob mice were observed by measuring the serum levels of glucose, insulin, lipid components, and the kidney levels of reactive oxygen species (ROS), MDA+HAE, GSH and also the enzyme activities involved in polyol pathway. Cheonghyul-san lowered the levels of serum glucose and insulin in a dose dependent manner. Total cholesterol, triglyceride and free fatty acid levels were decreased, while the HDL-cholesterol level was increased, in Cheonghyul-san treated groups. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the ob/ob mice, whereas those were inhibited in the Cheonghyul-san-administered groups. Cheonghyul-san inhibited the generation of ROS in the kidney. Finally, MDA+HAE level was increased and the GSH level was decreased in the ob/ob mice, whereas those were improved in the Cheonghyul-san-administered groups. The results suggested that Cheonghyul-san exerted the antidiabetic and antihyperlipidemic activities by regulating the activities of polyol pathway enzymes, scavenging ROS, regulating the MDA+HAE and GSH levels in the ob/ob mice.

• 대한화장품학회지
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• 제36권4호
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• pp.271-280
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• 2010

자외선(UV)은 인체 피부 손상의 주된 요인이고 UV에 대한 피부의 초기 반응들은 홍반과 부종을 포함한다. 화장품에 사용할 효과적인 자외선 방어제를 발굴하기 위하여 세포수준의 효능평가법으로 다양한 식물 추출물들을 검색하였다. 시험된 총 38종의 식물 추출물들 중 제주조릿대(Sasa quelpaertensis), 접시꽃(Althaea rosea), 관중(Dryopteris crassirhizoma)에서 유래한 3종의 식물 추출물이 배양된 인체 표피 멜라닌세포에서 멜라닌 생성뿐만 아니라 UVB의 세포독성을 경감시켰다. 이 식물 추출물들의 항염증 작용은 동물 실험에서도 평가하였다. 대조 크림 혹은 식물 추출물을 1 % 함유한 시험 크림을 C57BL/6 쥐의 귀에 혹은 SKH?1 무모쥐의 등에 UVB 조사 전과 후에 도포하였다. UVB 노출에 의한 영향으로서 부종 생성은 귀의 두께 변화를, 홍반 생성은 등 피부의 붉기의 변화를 측정하여 추정하였다. 3종류의 시험 크림 모두 두 실험 모두에서 항염증효과를 보였다. 제주조릿대, 접시꽃, 또는 관중 추출물을 함유한 크림은 자외선 조사 4일째 부종 반응을 각각 53.8 %, 56.4 %, 31.1 % 경감시켰다. 또한 이 크림들은 자외선 조사 2일째 홍반 생성을 각각 45.7 %, 34.1 %, 20.5 % 억제시켰다. 본 연구는 화장품에 함유된 특정 식물 추출물들이 자외선 과다노출로 야기되는 피부 염증을 완화시켜 줄 수 있음을 시사하였다.

• 생명과학회지
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• 제8권3호
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• pp.257-262
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• 1998

발암물질인 benzo(a)pyrene(B(a)P)이 암세포에 대한 세포성 방어능을 나타내는 자연살해(natural killer(NK))세포 활성에 미치는 영향을 관찰하기 위하여 사료를 C57BL/6마우스에 노출시킨 후 NK세포활성을 MTT시험법을 이용하여 측정하였다. 정상 및 시료노출군 마우스의 NK세포활성은 표적세포인 Yac-1세포에 대한 사멸세포백분율(percent dead cell)로서 측정하였다. 정상마우스의 비장세포에 B(a)P를 노출시켰을 때 노출군들은 시료농도에 따른 활성도에 큰 변화는 없었으니 낮은 농도($2.5^{\mu}g/ml$)의 노출에서도 정상에 비하여 현격히 낮은 활성치를 보였다. 정상마우스에 시료를 투여한 후에 분리한 비장세포의 NK세포활성치는 E/T ratio 200/1에서, 1회 투여보다는 2-3회 투여군에서 유의한 감소를 나타냄으로서 반복노출의 효과를 보였다. 본 시험을 통하여 B(a)P는 마우스 면역계에 미치는 효과중 암형성에 지대한 영향을 주는 NK세포활성을 크게 저하시킴을 알 수 있었다.

• 대한한의학방제학회지
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• 제19권1호
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• pp.219-231
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• 2011

Objectives : This study was designed to determine the effects of the GGEx18 ethyl acetate fraction(EF) on body weight gain, feeding efficiency ratio, and obesity-related factors in plasma as well as histology of liver and adipose tissues using high fat diet-fed male C57BL/6N obese mice. Methods : 8 weeks old, high fat diet-fed obese male mice were divided into 5 groups: C57BL/6N normal, control, EF(1), EF(2) and EF(3). After mice were treated with EF for 9 weeks, we measured body weight gain, food intake, feeding efficiency ratio, fat weight, plasma leptin and lipid levels. We also analysed histology of liver and adipose tissues on high fat diet-fed male C57BL/6N obese mice. Results : Compared with control, EF-treated mice had significantly lower body weight gain and feeding efficiency ratio. Consistent with the effects on body weight gain, EF significantly decreased the adipose tissue weight compared with control. Consistent with the effects on feeding efficiency ratio, EF significantly decreased plasma leptin concentrations compared with control. EF reduced the size of adipocytes as well as hepatic lipid accumulation compared with control. EF seems to be safe since not only the plasma levels of ALT and AST are within the normal range, but also EF did not show any toxic effects on organs. EF(3) was most effective among EF(1), EF(2), and EF(3) at doses of 25, 50, and 100 mg/kg, respectively. Conclusions : These results demonstrate that EF effectively reduces body weight gain, feeding efficiency ratio in high fat diet-fed obese mice, leading to the modulation of obesity. In addition, EF decreases the size of adipocytes and improves plasma lipids and controls hepatic lipid accumulation, suggesting that EF may act as a therapeutic agent for obesity.

• Journal of Acupuncture Research
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• 제27권5호
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• pp.59-68
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• 2010

목적 : 최근 한의학에서 널리 사용되며, 신경계 질환에도 응용되고 있는 봉약침의 농도의존적 효과를 알아보기 위하여, 대표적인 신경 퇴행성 질환인 파킨슨병의 동물모델을 통해 세포보호효과와 세포사멸 및 신경염증 기전을 관찰하였다. 방법 : C57BL/6 mice에 신경독소인 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine(MPTP)를 4번 복강내 주입하여 중뇌의 흑질 도파민 신경세포를 파괴하여 Parkinson 질병동물 모델을 만든 후, 2개의 군에는 마지막 MPTP 투여 2시간 후에 1차, 그 후로 48시간이 지날 때마다 양측 신수에 각각 0.06mg/kg 농도와 0.6mg/kg 농도의 봉약침을 시행하여 총 4회 시술한 후, 도파민 세포를 측정하는 TH 면역조직 화학법을 통해 세포의 보존 정도를 관찰하고, 세포사멸과 관련된 양상을 확인하기 위하여 Caspase 3, 신경염증과 관련된 양상을 확인하기 위하여 iNOS의 발현여부를 면역 조직화학법을 이용하여 관찰하였다. 결과 : 관찰결과 MPTP 투여 후 MPTP 투여군의 흑질의 도파민 세포 수는 감소하였으나 0.6mg/kg 봉약침을 투여한 경우에는 유의성 있게 세포 수가 유지되었다. Caspase-3와 iNOS 발현억제 실험에서 0.6mg/kg 봉약침군은 MPTP 투여군과 0.06mg/kg의 봉약침군과 비교하여 Caspase-3, iNOS 발현을 유의하게 억제하였다. 결론 : 봉약침은 MPTP 투여로 인한 신경세포 손상에 대하여 농도에 따라 세포사멸 기전과 신경염증 기전을 억제함으로 신경세포를 보호하는 것으로 추정되며, 추후 적절한 경혈점 및 최적의 봉약침 농도를 찾는데 지속적인 연구가 필요할 것이다.

• 약학회지
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• 제35권3호
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.292-297
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• 1994

This study was conducted to investigate antigenic potential of DA-3030, a recombinant human granulocyte-colony stimulating factor, in guinea pigs and mice. In the active systemic anaphylaxis test, the guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone did not show any anaphylactic reaction. In the homologous passive cutaneous anaphylaxis reaction, anti-DA-3030 antibody was not detected in guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone. On the other hand, the guinea pigs sensitized with 12.5 $\mu\textrm{g}$/heed of DA-3030 incorporated in Freund's complete adjuvant(FCA) or 1 mg/head of ovalbumin incorporated in FCA showed anaphylactic reaction. Anti-DA-3030 antibody was also detected in those guinea pigs. In immunodiffusion test using the sera sensitized with DA-3030 incorporated in FCA, precipitating antibodies were detected only in the sera sensitized with DA-3030 or DA-3030 incorporated in FCA showed. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone, none of the sera showed positive reaction. But sera of the animals immunized with 12.5 $\mu\textrm{g}$/head of DA-3030 incorporated in aluminum hydroxide gel(Alum) or 5 $\mu\textrm{g}$/head of ovalbumin incorporated in alum showed positive PCA reaction. DA-3030 did not cause anaphylactic shock or passive cutaneous anaphylaxis in guinea pigs and mice when given alone although DA-3030 incorporated in FCA or Alum induced anaphylactic shock and passive cutaneous anaphylaxis. From these results, it may be concluded the DA-3030 does not induce systemic allergic reaction when administered alone in its clinical use.

• Molecules and Cells
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• 제25권3호
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• pp.358-367
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• 2008

The embryonic germ cell (EGCs) of mice is a kind of pluripotent stem cell that can be generated from pre- and post-migratory primordial germ cells (PGCs). Most previous studies on DNA methylation of EGCs were restricted to 12.5 days post coitum (dpc). This study was designed to establish and characterize murine EGC lines from migrated PGCs as late as 13.5 dpc and to estimate the degrees of methylation of their imprinted genes as well as of the non-imprinted locus, Oct4, using an accurate and quantitative method of measurement. We established five independent EGC lines from post migratory PGCs of 11.5-13.5 dpc from C57BL/6 ${\times}$ DBA/2 F1 hybrid mouse fetuses. All the EGCs exhibited the typical features of pluripotent cells including hypomethylation of the Oct4 regulatory region. We examined the methylation status of three imprinted genes; Igf2, Igf2r and H19 in the five EGC lines using bisulfite genomic sequencing analysis. Igf2r was almost unmethylated in all the EGC lines irrespective of the their sex and stage of isolation; Igf2 and H19 were more methylated than Igf2r, especially in male EGCs. Moreover, EGCs derived at 13.5 dpc exhibited higher levels of DNA methylation than those from earlier stages. These results suggest that in vitro derived EGCs acquire different epigenotypes from their parental in vivo migratory PGCs, and that sex-specific de novo methylation occurs in the Igf2 and H19 genes of EGCs.