• Biomolecules & Therapeutics
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• 제31권6호
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• pp.629-639
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• 2023

Cardiovascular diseases (CVDs) are the most common cardiovascular system disorders. Cellular senescence is a key mechanism associated with dysfunction of aged vascular endothelium. Hyaluronic acid and proteoglycan link protein 1 (HAPLN1) has been known to non-covalently link hyaluronic acid (HA) and proteoglycans (PGs), and forms and stabilizes HAPLN1-containing aggregates as a major component of extracellular matrix. Our previous study showed that serum levels of HAPLN1 decrease with aging. Here, we found that the HAPLN1 gene expression was reduced in senescent human umbilical vein endothelial cells (HUVECs). Moreover, a recombinant human HAPLN1 (rhHAPLN1) decreased the activity of senescence-associated β-gal and inhibited the production of senescence-associated secretory phenotypes, including IL-1β, CCL2, and IL-6. rhHAPLN1 also downregulated IL-17A levels, which is known to play a key role in vascular endothelial senescence. In addition, rhHAPLN1 protected senescent HUVECs from oxidative stress by reducing cellular reactive oxygen species levels, thus promoting the function and survival of HUVECs and leading to cellular proliferation, migration, and angiogenesis. We also found that rhHAPLN1 not only increases the sirtuin 1 (SIRT1) levels, but also reduces the cellular senescence markers levels, such as p53, p21, and p16. Taken together, our data indicate that rhHAPLN1 delays or inhibits the endothelial senescence induced by various aging factors, such as replicative, IL-17A, and oxidative stress-induced senescence, thus suggesting that rhHAPLN1 may be a promising therapeutic for CVD and atherosclerosis.

• Applied Microscopy
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• 제38권1호
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• pp.21-28
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• 2008

식충식물의 잎은 결핍된 양분을 보충하기 위해 곤충을 유인하여 포획할 수 있는 포충엽으로 변형된다. 식충식물 표피조직에 발달하는 분비모는 특수한 모용으로 곤충포칙에 필요한 성분을 분비하여 먹이를 소화하고 흡수할 수 있도록 발달한다. 본 연구에서는 긴잎끈끈이주걱 (Drosera anglica Huds.) 엽신에 발달하는 여러 유형의 분비모 발달양상을 주사 및 투과전자현미경적인 방법으로 연구하였다. 포충엽 발달초기 엽원기 단계에서는 엽신이 접혀진 상태로 분화하여 상피조직은 노출되지 않으며, 하피조직에는 비분비모 및 모용원기들이 발달한다. 또한, 엽연이 접힌 상태로 어린 잎의 단계를 거치나 엽연에 발달하는 분비모는 기저부위만 노출되고 이들의 두정부위 및 다른 유형의 분비모들은 접힌 내부에 발달하여 보호된다. 병세포가 매우 길게 신장하는 엽연의 분비모에는 곤충 포획 시 가장 신속하게 식충의 기작을 수행하기 위한 세포수준에서의 구조분화도 수반된다. 엽연에는 긴 병세포를 지닌 약 $2.2{\sim}5.1\;mm$의 globose 분비모 (Type I)가 발달한 반면, 엽신의 내부로 갈수록 병세포는 점진적으로 짧아져 중앙 부위에서는 약 $200{\sim}300\;{\mu}m$의 짧은 병세포를 형성한다. 두정부위(head)는 2층의 분비세포, 중간의 장방형의 유세포층, 중앙의 가도관으로 구성된다. 엽연 분비모에 곤충이 접촉되면 이들 분비모는 매우 빠르게 움직이며 엽정 부위를 향축면으로 움직여 엽신이 곤충을 포위하게 하여 두정부위에서 분비된 점액성 물질로 곤충을 소화 흡수시키는 능동적인 식충의 기작을 수행한다. 또한, 엽신의 내부에는 분비물질을 분비하는 압핀형 두정부위와 짤은 병세포(ca. $15{\sim}30\;{\mu}m$)로 이루어진 분비모(Type II가 발달한다. 또 다른 유형은 병세포가 형성되지 않는 약 $25{\sim}40\;{\mu}m$의 6세포성분비모(Type III)로 상피 및 하피, 엽병, Type I 및 Type II 병세포 표면에 이르기까지 분포하며 위 두 유형의 분비모와 함께 활발한 분비활동을 수행한다. Type III 분비모의 경우, 분비물질은 정단의 두 세포에서만 방출되었다. Type I, II, III 두정부위 분비세포 세포벽에는 큐티클 층이 얇게 발달하며, 세포질 내에는 골지체, 소포체, 분비소낭 등의 막성계 세포내소기관 및 전자밀도가 높은 입자 등이 잘 발달하였다.

• Near Infrared Analysis
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• 제2권1호
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• pp.59-66
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• 2001

Nowadays, medical diagnostics is efficiently supported by clinical chemistry and near infrared spectroscopy is becoming a new dimension, which has shown high potential to provide valuable information for diagnosis. The investigation was carried out to study the influence of mammary gland inflammation, called mastitis, on cow´s milk spectra and milk composition measured by near infrared spectroscopy (NIRS). Milk somatic cell counts (SCC) in milk were used as a measure of mammary gland inflammation. Naturally occurred variations with milk composition within lactation and in the process of milking were included in the experimental design of this study. Time series of unhomogenized, raw milk spectral data were collected from 3 cow along morning and evening milking, for 5 consecutive months, within their second lactation. In the time of the trial, the investigated cows had periods with mammary gland inflammation. Transmittance spectra of 258 milk samples were obtained by NIRSystem 6500 spectrophotometer in 1100-2400 nm region. Calibration equations for the examined milk components were developed by PLS regression using 3 different sets of samples: samples with low somatic cell count (SCC), samples with high SCC and combined data set. The NIR calibration and prediction of individual cow´s milk fat, protein, and lactose were highly influenced by the presence of mil samples from animals with mammary gland inflammation in the data set. The best accuracy of prediction (i.e. the lower SEP and the higher correlation coefficient) for fat, protein and lactose was obtained for equations, developed when using only “healthy” samples, with low SCC. The standard error of prediction increased and correlation coefficient decreased significantly when equations for low SCC milk were used to predict examined components in “mastitis” samples with high SCC, and vice versa. Combined data set that included samples from healthy and mastitis animals could be used to build up regression models for screening. Further use of separate model for healthy samples improved milk composition measurement. Regression vectors for NIR mild protein measurement obtained for “healthy” and “mastitic” group were compared and revealed differences in 1390-1450 nm, 1500-1740 nm and 1900-2200 nm regions and thus illustrated post-secretory breakdown of milk proteins by hydrolytic enzymes that occurred with mastitis. For the first time it has been found that monitoring the spectral differences in water bands at 1440 nm and 1912 nm could provide valuable information for inflammation diagnosis.

• 생명과학회지
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• 제24권10호
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• pp.1070-1077
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• 2014

에스트로겐 수용체 알파에 높은 친화성을 갖고 있는 수용체 촉진제인 propyl pyrazole triol (PPT)을 성체 수컷 생쥐에 이틀 간격으로 8일, 16일, 24일 동안 피하주사방법으로 투여하였다. 현미경으로 부속 생식샘인 전립샘과 정낭, 그리고 간의 조직학적 변화를 관찰하였다. 대조군에 비해, PPT 투여군의 체중과 생식샘의 무게는 감소하였지만 간의 무게는 오히려 증가하였다. PPT 투여군에서 전립샘과 정낭의 내강 면적이 축소되었다. PPT 투여군 전립샘의 상피세포 높이는 대조군에 비해 증가하였다. 정낭의 경우, 상피세포의 핵 위쪽 세포질에 있는 분비 소포가 대조군에서는 나타났지만, PPT 군에서는 잘 관찰되지 않았다. 대조군에 비해 PPT 투여군 간의 굴모양혈관 직경이 투여횟수에 누적적으로 147.0%, 198.7%, 223.3% 의 비율로 증가되었다. 이런 결과는 PPT 투여로 인해 생식기관과 간의 조직 구조가 영향을 받으며, 그 조직학적 변화는 투여회수에 따라 더 심하게 나타났다. 결론적으로 이틀에 한번씩 투여한 PPT에 의해서 생식력 변화와 관련 있는 생식기관의 상피세포 높이 변화가 일어났고, 내강의 크기가 위축되었다. 간의 굴모양혈관 직경이 급격하게 확대되었으며, 이 결과는 간의 무게 증가로 이어졌다.

• Parasites, Hosts and Diseases
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• 제30권4호
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• pp.309-322
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• 1992

요꼬가와흡충(Metagonimus yokogawai)이 숙주의 장에 기생하여 성숙하는 동안 숙주에게 항체생성을 유도하는 물질이 충체 조직내 분포하는 부위를 규명하고, 특정 성숙기간 동안 숙주로부터 생성된 항체의 면역반응 정도를 확인하기 위하여 요꼬가와흡충 퍼낭유충을 실험고양이에 감염시키고, 성숙단계별 충체와 면역항체를 얻었다. 충체의 조직항원과 면역항체를 반응시키고 황금입자를 표지하여 황금입자가 보지된 조직항원은 전자현미경으로 관찰하였다. 표지된 환금입자의 밀도에 따라 조직성분의 항원성을 측정한 결과 감염군 면역항체를 반응시킨 경우 숙주 체내에서 4주와 8주 그리고 12주 동안 성장한 충체 조직항원은 표피충의 표피합포체에 다소 높은 밀도의 황금입자가 표지되어 항원성이 강한 것으로 관찰되었다. 16주와 20주간 성장한 충체의 조직항원은 표피층의 액포에 황금입자의 표지가 현저하게 증가하여 16주이상 성장한 충체 표피충의 액포를 구성하는 물질이 항원성이 강한 것으로 관찰되었다. 유연조직의 분비과립은 충체의 감염기간에 상관없이 일정하게 항원성이 나타났으나 맹관상피는 16주 이상 성장한 충체부터 항원성이 다소 미약해졌다. 숙주가 생성하는 요꼬가와흡충에 대한 항체는 감염 후 4주부터 12주사이에는 충체의 성장기간과 상관없이 일정 량으로 숙주의 혈청 내에 존재하는 것으로 생각되었으나 16주 이후에는 다소 감소하는 것으로 생각되었다.

• 대한수의학회지
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• 제43권4호
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• pp.541-549
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• 2003

The present study investigated the effects of aging on Leydig cells of Sprague Dawley rats. Rats of 3, 6, 12 and 18 months of age were used. Testes of rat were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in epon-araldite. Using $1{\mu}m$ sections stained with methylene blue, qualitative and quantitative morphological studies were performed. Testis incubations were used to determine luteinizing hormone (LH; 100 ng/ml) stimulated testosterone secretory capacity per testis in vitro. Testosterone levels in the incubation medium, and testosterone and luteinizing hormone levels in serum of these four groups of rats were determined by radioimmunoassay. Morphological studies revealed that Leydig cells were more abundant in the testis interstitium at 6, 12 and 18 months when compared with 3 months. The volumes of Leydig cells per testis was significantly higher, at 6, 12 and 18 months of age than those at 3 months. The number of Leydig cells per testis was doubled at 6, 12 and 18 months of age compared with 3 months. The average volume of a Leydig cell was not significantly different between 3 and 6 months of age, however, at 12 and 18 months a significantly lower value was observed. LH-stimulated testosterone production per testis in vitro was reduced by 45% at 6 months of age compared with 3 months; a further significant reduction was observed at 12 and 18 months. Serum testosterone and LH levels were not significantly different between 3 and 6 months of age but at 12 and 18 months a significantly lower value was observed in both groups for these hormones. These results showed that signs of aging are apparent in Leydig cells of Sprague Dawley rats at 12 months of age.

• 대한수의학회지
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• 제32권4호
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• pp.497-510
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• 1992

This study was attempted to comparative investigate the types and regional distribution of the endocrine cells in several vertebrates immunohistochemically using seven antisera. From carp pancreas could be observed 4 types which are insulin-, glucagon-, som- and BPP-immunoreactive cells. Insulin-immunoreactive cells were mainly distributed at the periphery and a few cells occupied the central region of the islets. Glucagon-immunoreactive cells were distributed at the periphery of the islets, and som - and BPP-immunoreactive cells were located at the central region. From frog pancreas could be observed 4 types which are insulin-, glucagon-, som- and BPP-immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the islets. Som-immunoreactive cells were distributed at the periphery of the islets, and glucagon- and BPP-immunoreactive cells were found as single cell or as small groups located between the pancreatic acini. From snake pancreas could be observed 3 types which are insulin-, glucagon- and som -immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the small islets, and they also were scattered at the periphery of the large islets. Glucagon-immunoreactive cells were distributed at the periphery of the islets, whereas som-immunoreactive cells were occupied the central region. From Ogolgae pancreas could be observed 4 types which are insulin-, glucagon-, som-and BPP-immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the small islets, but at the periphery of the large one. Glucagon- immunoreactive cells were distributed at the periphery of the small islets and in the large islets showed scattering entired. Som-immunoreactive cells were distributed at the periphery of the small islets and in the large islets were located at the central region. A small numbers of BPP-immunoreactive cells were located at the periphery of the small islets and the exocrine regions. From the pancreas of the Korean native goat could be observed 6 types which are insulin-, glucagon-, som-, BPP-, 5-HT- and porcine-CG-immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the islets. Som-immunoreactive cells were located at the periphery of the islets, but a tew were scattered at the central region of islets and in the epithelium of the secretory duct. Glucagon-, BPP-, 5-HT- and porcine CG-immunoreactive cells were distributed at the periphery of the islets. These findings indicated that the regional distribution patterns and cell types of pancreatic endocrine cells in vertebrates varies considerably among phylogenetically different vertebrates.

• Applied Microscopy
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• 제36권4호
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• pp.271-277
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• 2006

본 연구에서는 흰쥐 발바닥에 complete Freund's adjuvant(CFA)를 투여한 후 염증부위에서 관찰되는 비만세포의 분포양상 및 미세구조를 zinc selenium autometal-lography$(AMG^{ZnSe})$로 염색한 후 광학 및 전자현미경으로 관찰하였다. 광학현미경으로 관찰된 CFA투여에 의한 염증성 반응은 피하층에 국한되어 있었으며, zinc함유 비만세포는 주로 피하조직의 혈관 주변에 분포하고 있었다. AMG 염색에 의한 흑갈색을 띠는 과립이 세포질을 채우고 있었으며, 많은 비만세포의 경우 세포막이 파괴되어 과립이 주위조직으로 퍼져 관찰되었다. 전자현미경으로 관찰된 zinc함유 비만세포는 불규칙한 외형을 띠었으며, 표면에는 작은 돌기들이 관찰되었다. AMG염색에 따른 은입자(silver grains)가 세포질내 과립에 국한되었으며, 높은 전자 밀도의 동질성 기질로 구성되어 있었으며, 경계막으로 싸여 있었다. 세포질내 세포소기관 중에서 특히 골지 복합체가 잘 발달되어 있었으며, 형질내세망과 사립체는 비교적 적게 가지고 있었다. 이들 세포의 핵은 비교적 작고 둥글고, 세포의 중심부에 위치하였으며, 이질염색질로 이루어져 있었다.

• Applied Microscopy
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• 제22권2호
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• pp.1-14
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• 1992

This experiment was performed to study the morphological responses of the parafollicular cells of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The head and neck region of the rat, under sodium thiopental anesthesia, was exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80 cm, and the dose rate was 200 rads/min. The rate of experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Pieces of the tissue taken from the thyroid gland were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH 7.3), and in 1% osmium tetroxide (0.1M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. Two types of the parafollicular cells, according to their electron densities, were found, i. e., light cells and dark cells. 2. Three types of the parafollicular cells, according to their sizes of secretory granules were found, i.e., small granule cells ($85nm{\pm}20.1;64{\sim}102nm$), medium granule cells ($120nm{\pm}26.5;77{\sim}179nm$), and large granule cells ($165nm{\pm}25.7;128{\sim}189nm$). 3. The differential ultrastructural changes of the cells according to their cell types, i.e., dark and light cell, or small, medium and large granule cells, were hardly observed in the time and dose range covered by this study. 4. The morphological changes of the parafollicular cells were not pronounced after exposure to 3,000 rads of X-ray. 5. Swollen cisternae of the granular endoplasmic reticulum and partial cytolysis were observed after exposure to 6,000 rads of X-ray. 6. Above results suggest that the parafollicular cells showed the alterations of mitochondrial and granular endoplasmic reticular swelling, and partial cytolysis, but only in doses of 6,000 rads.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.54-54
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• 2003

Prostate gland contains neuroendocrine cells (PNECs) are playing important roles in physiological and pathophysiological processes of the prostate gland. Here, we investigated the role of purinoceptors in PNECs freshly isolated from rat ventral prostate (RPNECs) that show immunoreactivity to chromogranin A. Fura-2 ratiometry revealed that ATP evokes both fast Ca$\^$2+/ influx and store Ca$\^$2+/ release in RPNECs. A whole-cell patch clamp study demonstrated fast inactivating cationic current activated by ATP or by ${\alpha}$,${\beta}$-MeATP, which was blocked by ATP-TNP. The activation of P2X inward current was tightly associated with a sharp increase in [Ca$\^$2+/]$\sub$c/. The presence of P2X1/3 subtypes were proved by RT-PCR analysis. For the stored Ca$\^$2+/ release, ATP and UTP showed similar effects, suggesting the dominant role or P2Y2 subtypes, also confirmed by RT-PCR. Both P2X (${\alpha}$,${\beta}$-MeATP) and P2Y (UTP) stimulation induced changes in the cell morphology (initial shrinkage and blob formation on the surface) reversibly. Exocytotic membrane trafficking events were monitored with the membrane-bound fluorescent dye, FM1-43 using confocal microscopy. In spite of the similar Ca$\^$2+/ responses, UTP was far less effective in triggering exocytosis than ${\alpha}$,${\beta}$ -MeATP. Since serotonin is reportedly stored in the secretory granule of PNECs, we directly examined whether the aforementioned agonists elicit release of serotonin using carbon fiber electrode-amperometry. In accordance with the results of FM1 -43 experiments, ${\alpha}$,${\beta}$-MeATP efficiently evoke serotonin secretion while not with UTP. In summary, the P2X-mediated Ca$\^$2+/ influx plays crucial roles in the exocytosis of RPNECs. Although a global increase in [Ca$\^$2+]$\sub$c/ might be related with the morphological changes, a sharp rise of [Ca$\^$2+/]$\sub$c/ in the putative sub-plasmalemmal ‘microdomains’ might be a decisive factor for the exocytosis.