• Journal of Ginseng Research
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• 제36권1호
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• pp.78-85
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• 2012

Endothelial progenitor cells (EPC) are a population of cells that circulate in the blood stream. They play a role in angiogenesis and, therefore, can be prognostic markers of vascular repair. Ginsenoside $Rg_3$ prevents endothelial cell apoptosis through the inhibition of the mitochondrial caspase pathway. It also affects estrogen activity, which reduces EPC senescence. Sun ginseng (SG), which is heat-processed ginseng, has a high content of ginsenosides. The purpose of this study was to investigate the protective effects of SG on senescence-associated apoptosis in EPCs. In order to isolate EPCs, mononuclear cells of human blood buffy coats were cultured and characterized by their uptake of acetylated low-density lipoprotein (acLDL) and their binding of Ulex europaeus agglutinin I (ulex-lectin). Flow cytometry with annexin-V staining was performed in order to assess early and late apoptosis. Senescence was determined by ${\beta}$-galactosidase (${\beta}$-gal) staining. Staining with 4'-6-Diamidino-2-phenylindole verified that most adherent cells (93${\pm}$2.7%) were acLDL-positive and ulex-lectin-positive. The percentage of ${\beta}$-gal-positive EPCs was decreased from 93.8${\pm}$2.0% to 62.5${\pm}$3.6% by SG treatment. A fluorescence-activated cell sorter (FACS) analysis showed that 4.9% of EPCs were late apoptotic in controls. Sun ginseng decreased the apoptotic cell population by 39% in the late stage of apoptosis from control baseline levels. In conclusion, these results show antisenescent and antiapoptotic effects of SG in human-derived EPCs, indicating that SG can enhance EPC-mediated repair mechanisms.

• 사상체질의학회지
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• 제12권2호
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• pp.171-183
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• 2000

1. Back ground and purpose An experimental study has done to examine the effect of defense and cure gastric mucasal damage of Mokdan-pijihwang-tang. 2. Methods Mice had intragastric injected with MJ extract before indomethacin treatment which induces homorrhage infarct and erosion artificially. Degree of lipid peroxidation, general morphology, change of mucous cell, the distribution of PNA, ICAM and distribution of apoptotic cell were objected. (Abbreviation) MJ : Mokdanpijihwang-tang, PNA : Peanut Agglutinin, ICAM : Intercellular Adhesion Molecule 3 Results 1) The degree of lipid peroxidation in INDO-group had increased conspicuously than control group. But the degree of lipid peroxidation in MJ-group had decreased than INDO-group and these decline had probability. 2) After indomethacin treatment, hemorrhage infarct and erosion had increased in stomach body. But in MJ-group, the configuration is normal, except the group intragastric injected with MJ extract at hour 24 before indomethacin treatment. 3) Surface mucous cell and neck mucous had disappeared in INDO-group. But in MJ-group tormal distribution had shown like control group except the group intragastric injected with MJ extract at hour 24 before indomethacin treatment. 4) PNA positive reaction had not shown in INDO-group. But medium PNA positive reaction had shown In Mj-group. 5) ICAM positive reacted cell had shown in INDO-group. The decrease of ICAM positive cell were shown than INDO-group. 6) A number of apoptotic cell was distributed in hemorrhagic erosion. A few number of apoptotic cell was distributed in MJ-group except some surface mucous. 4. Conclusion These results suggest that MJ has an effect on cure of gastric mucosal damage.

• 한국동물학회지
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• 제39권1호
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• pp.26-35
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• 1996

쥐의 외측 망상핵으로부터 소뇌충부 제6엽 내의 각 소엽으로 신경 경로를 WGA-HRP를 이용한 역행수송법을 써서 조사하였다. 표지된 신경세포는 양측의 외측 망상핵에 모두 존재하였으나, 동축의 경우에 편중되어 있었다. 동측 또는 대측의 외측 망상핵의 큰 세포구획(magnocellular division)에서 국소순적으로 배열이 관찰되었는데, a소엽에서 c소엽으로의 투사가 동측의 큰 세포구획에서 등쪽에서 배쪽으로 분포양상을 보였으며, 대측의 큰세포구획에서는 다소 머리측에서 꼬리측 절편으로의 분포 양상을 보였다. 그 외 동측 또는 대측의 작은세포구획(parvocellular division) 및 삼차밑구획(subtrigeminal division)에서의 표지된 신경세포의 수는 극히 적었다. 한편 외측 망상핵으로부터 소뇌층부의 제6엽 a소엽/b소엽으로서의 투사에 관한 컴퓨터를 이용한 삼차원 재구성은 각 경우에 있어서 상당량의 투사의 중첩이 존재함을 보여주고 있다.

• 한국동물학회지
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• 제38권4호
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• pp.514-522
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• 1995

생쥐 정자의 수정능력 획득과정 및 첨체반응을 전후로 정자표면과 albumin의 상호작용을 조사하였다. 부정소 정자를 체외에서 배양하는 과정에서 정자표면에서 이탈된 단백질의 분석과 함께 FITC-bovine serum albumin으로 정자를 형광염색하여 정자표면에 대한 albumin의 결합양상의 변화를 조사하였다. 90분간 정자를 배양한 수 정자를 제거한 후 농축한 배양액내에 정자 또는 부정소액에서 기원한 여러종의 단백질가 함께 albumin이 다량으로 발견되었다 정자의 체외배양 과정에서 일어나는 albumin의 이탈은 배양액내의 $Ca^2$+과 무관하게 일어났다. BSA-FITC는 정소내 정자의 두부표면에 미약하게 결합한 반면 미부부정소 정자의 첨체표면에는 다량 결합하였다. $Ca^2$+-ionophore인 A23187으로 첨체반응을 유발한 정자의 두부 표면에서는 후첨체부위만이 강하게 염색되었다. 이러한 결과는 정소 및 웅성 생식수관니에서 정자표면에 부착된 albumin이 자성 생식수관을 거치는 동안 이탈됨을 시사하며 이러한 현상은 정자의 수정능력획득과 밀접한 관련이 있는 것으로 사료된다.

• 사상체질의학회지
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• 제16권2호
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• pp.84-98
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• 2004

1. Objectives Yangkyuksanhwa-tang is used mush in pruritus and dermatopathy of Soyangin. It is suggested this prescription is effective on atopy dermatitis. 2. Methods For observation of Yangkyuksanhwa-tang effected to atopic dermatitis, extract of Yangkyuksanhwa-tang has been dispensed to the stratum corneum of epithelium in dermatome of murine after making damage to its defense mechanism against fat and causing atopic dermatitis artificially. After that, the change in outer dermatome and minute mechanism of epidermis, the change of eosinophil, the change in distribution of soybean agglutinin, the change in distribution of fat and ceramide in stratum corneum, the change in inflammation in dermatome, the change of cell accrementition and apoptosis, and the effect on anaphylaxis and Staphylococcus aureus was observed. 3. Results After administration of Yangkyuksanhwa-tang, severe skin damage such as eczema and psoriasis, that was observed in the case of atopy dermatitis, was decreased and the increase of eosinophil in serum was suppressed. Lipid lamella was recovered, so epidermal demage was relieved. The distribution of HSP70 in the outer skin was decreased. Yangkyuksanhwa-tang suppressed activation of $NF-_{\kappa}B$ p50, induced CD11/18b not to be generated, and suppressed inflammatory response of skin. Anaphylaxis and groth of Staphylococcus aureus was suppressed. 4. Conclusions Yangkyuksanhwa-tang decreased skin damage of atopy dermatitis. It has antibiosis about Staphylococcus aureus, it can be medicinal substances on atopy dermatitis. In addition, it is possible that it can be medicinal substances on regional skin allergy.

• Biomolecules & Therapeutics
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• 제2권4호
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• pp.376-382
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• 1994

Repeated injections of low-doses of mercuric chloride in rats or mice induce polyclonal activation which includes the induction of anti-glomerular basement membrane (GBM) antibodies and circulating immune complex and it results in nephritis. Because this disease is autoimmune mediated disease resulted from immune dysfunction, immunomodulators are used to control the symptoms or to cure the disease. Irpex lacteus Fr. is a kind of new medicinal fungus. The polysaccharide fraction extracted from submerged fermentation of Irpex lacteus Fr. decreased the serum agglutinin, serolysin and IgM plaque forming cells in normal mice. The hitherto obtained clinical results suggested that it significantly improved the oligourea, edema, and hypertension in patients who have nephritis. To elucidate the action-mechanisms of Irpex lacteus Fr., we established the experimental model of HgCl$_2$induced polyclonal activation by intraperitoneal administrations of HgCl$_2$to mice. To assess the immunomodulating effect of Irpex lacteus fraction, we Investigated its effects on the mitogen induced proliferation and IgM PFC counts of splenic lymphocytes in mice during the treatment of HgCl$_2$. The Irpex lacteus polysaccharide reduced the abnormally increased mitogen induced Iymphocyte proliferation and IgM PFCs to almost normal levels. And the Irpex lacteus polysaccharides prevented the increasement of serum immunoglobulin level induced by HgCl$_2$. These data suggested that the Irpex lacteus polysaccharides might have the immunomodulating activity to prevent and /or improve the HgCl$_2$ induced autoimmune disease.

• 약학회지
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• 제41권4호
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• pp.421-432
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• 1997

A new lectin was partially purified from starfish,Asterina pectinifera by means of physiological saline extraction, salt fractionation, ion exchange chromatography and hy droxyapatite chromatography, and it was named APL. The biochemical properties of the APL were characterized. In addition, its effects on lymphocyte mitogenicity and cancer cell agglutinability were tested. The APL agglutinated nonspecifically human erythrocytes and rabbit blood cells. Agglutinability was decreased to 30% of control activity below pH 5 and above pH 9 and was relatively unstable at increasing temperatures above 60$^{\circ}C$. The activity was reduced by addition of two kinds of metal ions, $Ba^{2+},\;Mn^{2+}$ and chelating agent, EDTA. APL was proved to be glycoproteins containing 9% sugars. For carbohydrate specificity, it was found that the activity of APL was inhibited by D(+)-glucosamine, D(+)-galactosamine, stachyose, N-acetyl-galactosamine and methyl-${\alpha}$-D-galactopyranoside among 35 sugars tested. In amino acid composition, the contents of acidic amino acids such as aspartic acid and glutamic acid were relatively high. This result suggest that the isoelectric point would be in a lower range. APL was found that it promotes the division of human lymphocytes. APL was proved to be a potent agglutinin for cancer cells such as HeLa, L929 and L1210 cells. Significant changes on the HeLa cell surfaces affected by APL were observed under the electron microscope.

• 한국동물생명공학회지
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• 제37권2호
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• pp.130-135
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• 2022

Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO₂, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN₂ vapor. After thawing at 37℃ for 25 s, Isolate^® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.

• 한국임상수의학회지
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• 제30권6호
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• pp.442-448
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• 2013

개 정액 동결을 위한 glucose가 첨가된 glycerol-free TRIS 희석액을 개발하기 위해 glycerol-free TRIS내 알맞은 glucose의 양과 0.3 M glucose가 첨가된 glycerol-free TRIS내 정자의 적정 노출시간을 조사하였다. 여섯 마리의 수캐의 사출액을 0.04 M glucose가 첨가된 glycerol-free TRIS내에서 $4^{\circ}C$까지 100분 동안 냉각한 후 서로 다른 glucose농도 (0 M, 0,04 M, 0.1 M, 0.2 M, 0.3 M)의 glycerol-free TRIS에서 30분 동안 냉각하여 동결하였다. $37^{\circ}C$에서 25 초 동안 융해한 후 정자의 막 고유성과 첨단체 고유성을 검사하였다. 부가적으로 0.3 M glucose가 첨가된 glycerol-free TRIS내 정자의 적정 노출시간에 따른 정자의 동결 후 운동성, 생존성, DNA 고유성을 확인하였다. 막 고유성과 첨단체 고유성은 각각 6-carboxyfluoresceindiacetate(6-CFDA)/propidium iodide(PI) fluorescent staining와 Pisum sativum agglutinin conjugated-fluorescein isothiocyanate 방법에 의해 검사하였다. DNA 고유성은 terminal deoxynucleotidyl transferase dUTP nick end labeling로 염색하여 flow cytometry로 검사하였다. 0.2 M 또는 0.3 M glucose가 첨가된 glycerol-free TRIS에서 동결된 정자가 낮은 농도의 glucose가 첨가된 희석액에서 동결된 정자보다 막 고유성이 높게 나타났으며(p<0.05), 첨단체 고유성은 0.3 M 군에서 높게 나타났다(p<0.05). 운동성은 50 분 군에서 높게 나타났으나(p<0.05), DNA fragmentation index는 노출시간에 따라 차이가 없었다. 본 연구 결과 개정자가 0.3 M glucose가 첨가된 glycerol-free TRIS에서 $4^{\circ}C$, 50 분간 냉각 후 동결과 융해 후 더 높은 생존성을 나타냈다.

• 한국임상수의학회지
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• 제24권4호
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• pp.486-492
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• 2007

개 정액의 정장은 정자를 동결 보존하기전 정액에서 제거되어진다. 그러나 근래 개정장이 정자를 냉각하고 동결하는데 유효한 효과가 있는 것으로 보고되고 있다. 그러므로 본 연구에서는 정자를 냉각과 동결하기 전 정장을 정자 희석액에 첨가하여 정장이 정자의 생존성에 미치는 영향을 알아보고자 하였다. 건강한 4마리의 수캐로부터 정액을 채취 혼합한 후 300g에서 10분간 원심분리하여 얻은 상층액을 다시 원심분리하고 상층액을 회수하여 정장으로 사용하였다. 정자의 희석액으로는 Egg yolk-Tris(EYT)를 사용하였다. 다음과 같이 두 가지 실험을 실시하였다. 실험 1: 정장이 0, 20, 40, 80, 100%비율로 포함된 EYT배지에 정자를 희석하여 두 시간 동안 $4^{\circ}C$나 $25^{\circ}C$에서 보관하였다. 정자수는 $2{\times}10^8/ml$로 조정하였다. 실험 2: 전체적으로 9개 실험군으로 구분하여 4개 실험군은 정장이 20, 40, 80, 100%포함된 EYT배지에 각각 정자를 희석하여 $4^{\circ}C$까지 냉각한 후 EYT+1.2 M glycerol배지로 2차 희석하여 동결하였다. 나머지 4개의 실험군은 EYT배지에 정자를 희석하여 $4^{\circ}C$까지 냉각한 후 각각 정장이 20, 40, 80, 100%포함된 EYT+1.2 M glycerol배지로 2차 희석하여 동결하였다. 대조군으로서 정장을 포함하지 않은 EYT배지에 정자를 희석하여 $4^{\circ}C$까지 냉각한 후 EYT+1.2 M glycerol배지로 2차 희석하여 동결하였다. glycerol의 최종농도는 0.6M, 정장의 최종농도는 10, 20, 40, 50%이었으며, 정자수는 $1{\times}10^8/ml$로 조정하였다. 정자를 straw에 충진하고 분당 $25^{\circ}C$의 냉각율과 액체질소를 이용하여 동결하였다. 융해는 $38^{\circ}C$에서 1분간 실시하였다. 정자 운동성, 정자 형질막 보존성(생존력), 정자 첨단체의 보존성을 검사하여 정자의 생존성을 검증하였다. 정자의 운동성은 400배율에서 현미경으로 관찰하였으며 생존력과 첨단체 보존성은 각각 이중형광염색과 Pisum sativum agglutinin을 이용하여 검사하였다. 실험 1의 결과는 보존온도와는 관련없이 희석액에 정장을 첨가하는 것은 정장이 포함되지 않은 희석액과 비교하여 정자의 운동성을 향상시키지 못했다. 실험 2 결과, 동결 전 냉각상태에서 정장을 포함한 EYT+0.6 M glycerol배지에 희석되었던 정자가 정장을 포함하지 않은 EYT+0.6 M glycerol배지보다 높은 진행운동성을 보였다(P<0.05). 그러나, 동결 융해 후 정장을 포함한 EYT+0.6 M glycerol배지에 희석되었던 정자의 생존력은 정장을 포함하지 않은 EYT+0.6 M glycerol배지보다 낮거나 비슷하게 나타났다. 본 실험의 결과를 요약하면 비록 정장이 동결보호제가 포함되지 않은 EYT에 혼합되었을 경우는 냉각되어진 정자의 생존성을 향상시키지는 못했으나 동결보호제가 포함된 EYT에 정장을 혼합한 경우 정장은 정자의 냉각 후 정자의 운동성을 향상시켰다.