• 대한수의학회지
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• 제36권3호
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• pp.577-598
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Chronic test were conducted to determine those effects. The chronic test was conducted by dividing rats into 5 groups according to the type of drugs and dosages administrated as follows; the control(group A), and group B was given 25mg/kg caffeine orally once daily for 30 days, group C was given 50mg/kg caffeine orally once daily for 30 days, group D was given 25mg/kg caffeine and orally ferric chloride once daily for 30 days and group E was given 25mg/kg caffeine and 25mg/kg vitamin E once daily for 30 days. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group and malondiaidehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis) and fatty acid compositions in free fatty acids and phospholipids were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. Body weights of groups B, C, D and E were significantly decreased(p < 0.01) in comparison with that of the control in the chronic test. 2. The concentrations of serum glucose in groups B(124.5mg/dl), C(130.1mg/dl), D(122.1mg/dl), E(119.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(101.5mg/dl). But, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and A/G ratio in comparison to that of the control. 3. The concentrations of total cholesterol and HDL-cholesterol in serum of groups B(69.6, 53.4mg/dl), C(73.0, 56.3mg/dl), D(68.9, 51.1mg/dl) and E(68.2, 51.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(52.6, 38.8mg/dl). On the other hand, the concentrations of triglyceride in serum of groups B(45.0mg/dl), C(40.4mg/dl), D(33.8mg/dl) and E(47.2mg/dl) were significantly lower(p < 0.01) in comparison to that of the control(66.2mg/dl). There were no significant differences in the activities of ALT, AST and ALP in comparison to that of the control. 4. The concentrations of free fatty acid and phospholipid in serum of groups B(45.7, 154.4mg/dl), C(50.0, 167.2mg/dl), D(52.5, 148.4mg/dl) and E(41.1, 159.2mg/dl) were higher(p < 0.01) in comparison to that of the control(35.2, 125.3mg/dl). And the concentrations of the carbonyl group and malondialdehyde in serum of group D(1.82, 0.52nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(1.53nM/mg protein). 5. The concentrations of carbonyl group in total homogenate, mitochondrial and microsomal fraction of group D(1.45, 0.94, 1.67nM/mg protein) were significantly higher (p < 0.01) in comparison to the control(1.16, 0.66, 1.27nM/mg protein). And the concentrations of malondialdehyde in the total homogenate, mitochondrial and microsomal fraction of group D(6.70, 6.10, 1.36nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(5.17, 3.64, 0.68nM/mg protein). 6. As the analytical results of the fatty acid compositions of free fatty acid in serum, the proportions of stearic acid and arachidonic acid of groups B(16.52, 12.62%), C(17.52, 15.18%), D(19.73, 13.47%) and E(17.62, 13.28%) were significantly higher(p < 0.01) in comparison to the control(14.75, 7.88%), but the proportions of oleic acid and linoleic acid of groups B(12.97, 32.59%), C(10.88, 31.23%), D(12.37, 30.66%) and E(11.95, 32.41%) were significantly lower(p < 0.01) in comparison to the control(16.44, 35.12%). Otherwise, as the results of the fatty acid compositions of phospholipid in serum, the proportions of stearic acid and arachidonic acid of groups B(39.37, 16.39%), C(40.63, 17.83%), D(42.73, 15.39%) and E(39.16, 15.70%) were significantly higher(p < 0.01) in comparison to the control(37.74, 14.24%), but the proportions of oleic acid and linoleic acid of groups B(4.03, 14.38%), C(3.54, 12.38%), D(4.52, 11.68%) and E(4.29, 13.64%) were significantly lower(p < 0.01) in comparison to the control(5.53, 16.14%). 7. As the analytical results of the fatty acid compositions of free fatty acid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of oleic acid of groups B(7.8**, 8.73**, 6.88%) and C(6.89**, 7.75**, 6.58%) were lower(**:p < 0.01) in comparison to the control(8.67, 10.08, 7.81%), but the proportions of arachidonic acid of group C(22.62, 19.79, 23.71%) were significantly higher(p < 0.01) in comparison to the control(20.93, 18.47, 22.24%). And the proportions of palmitic acid of group D(25.95**, 26.16, 26.34**%) were significantly higher(**:p < 0.01) in comparison to the control(24.43, 25.42, 23.34%). In addition, the proportions of linoleic acid of group D(23.43, 25.02, 23.95%) were also significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). The proportions of stearic acid of group D(19.87, 19.76**%) in mitochondrial and microsomal fraction were lower(**:p < 0.01) in comparison to the control(21.01, 24.18%), and the proportions of stearic acid of group E(16.71*, 19.65**%) in mitochondrial and microsomal fraction were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(21.01, 24.18%), and the proportions of linoleic acid of group E(25.04, 29.20, 26.48%) in total homogenate, mitochondria and microsome were significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). 8. As the results of the fatty acid compositions of phospholipid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of palmitic acid of group D(17.58**, 18.78*, 18.23%**) were significantly higher(**:p < 0.01, *:p < 0.05) in comparison to the control(16.28, 17.22, 16.38%), and the proportions of stearic acid of group D(36.41, 37.23, 39.53%) were also significantly higher(p < 0.01) in comparison to the control(34.18, 34.16, 36.04%). But the proportions of oleic acid(3.41*, 3.11**, 3.12**%) and linoleic acid (18.03**, 15.79**, 14.74**%) of group D were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(oleic : 3.63, 3.72, 3.79%, linoleic : 20.03, 18.71, 18.48%). 9. In order to determine the oxidative damages to the protein in serum, mitochondrial and microsomal fraction of the rat liver, the patterns of the SDS-PAGE were identified, but the results of SDS-PAGE were not significantly different between the control and experimental groups.

• Nutrition Research and Practice
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• 제7권6호
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• pp.481-487
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• 2013

High-fat diet up-regulates either insulin resistance or triglycerides, which is assumed to be related to the expression of peroxisome proliferator-activated receptor (PPAR)-${\alpha}$ and PPAR-${\gamma}$. The beneficial effects of vitamin E on insulin resistance are well known; however, it is not clear if vitamin E with a high-fat diet alters the expression of PPAR-${\alpha}$ and PPAR-${\gamma}$. We investigated the effects of d-${\alpha}$-tocopherol supplementation on insulin sensitivity, blood lipid profiles, lipid peroxidation, and the expression of PPAR-${\alpha}$ and PPAR-${\gamma}$ in a high-fat (HF) diet-fed male C57BL/6J model of insulin resistance. The animals were given a regular diet (CON; 10% fat), a HF diet containing 45% fat, or a HF diet plus d-${\alpha}$-tocopherol (HF-E) for a period of 20 weeks. The results showed that the HF diet induced insulin resistance and altered the lipid profile, specifically the triglyceride (TG) and total cholesterol (TC) levels (P < 0.05). In this animal model, supplementation with d-${\alpha}$-tocopherol improved insulin resistance as well as the serum levels of TG and very-low-density lipoprotein-cholesterol (VLDL-C) (P < 0.05). Moreover, the treatment decreased the levels of malondialdehyde (MDA) in the serum and liver while increasing hepatic PPAR-${\alpha}$ expression and decreasing PPAR-${\gamma}$ expression. In conclusion, the oral administration of d-${\alpha}$-tocopherol with a high-fat diet had positive effects on insulin resistance, lipid profiles, and oxidative stress through the expression of PPAR-${\alpha}$ and PPAR-${\gamma}$ in a high-fat diet-fed male mice.

• Journal of Nutrition and Health
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• 제53권4호
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• pp.381-389
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• 2020

본 연구는 국내 비만과 고도비만 여성의 혈청 영양소 수준을 관찰하고자 시행하였다. 본 연구 결과를 통해 비만인과 고도비만인의 비만 예방 및 관리 지표로서의 혈청 영양소와의 관련성을 파악하고자 하였다. 본 연구의 대상자의 연령은 만18세 이상에서 59세 이하 성인 여성이며 서울 B 외과의원 비만클리닉에서 제공된 ID에 의하여 판별되는 2차 가공된 자료로 병원에서 환자의 동의를 받고 후향적으로 자료를 분석하여 총 44명을 대상자로 선정하였다. 본 연구는 BMI의 지표를 활용하여 BMI가 25.0-29.9 kg/㎡의 비만을 비만군, 30.0 kg/㎡ 이상의 고도 비만을 고도비만군으로 구분하여 비만군 23명, 고도비만군 21명을 분석하였다. 연구 대상자들의 신장과 체중, BMI, SMM, BFM, PFM을 체성분 분석을 통해 조사하였고, 혈중 생화학적 지표와 엽산, 비타민 B₁₂, 25(OH)D₃, 철, 페리틴 수준을 분석하였다. 비만군과 고도 비만군의 연령, 체중, BMI 및 체조성 중 체지방량, 체지방률은 비만군이 유의적으로 낮게 나타났으며 신장은 고도 비만군에서 유의적으로 낮게 나타났다. 생화학적 지표 검사결과 AST, ALT, 당화혈색소, 총 콜레스테롤, 중성지방은 대부분 정상 범위에 해당하였으며 두 군 사이의 유의적 차이가 나타나지 않았다. 비만과 관련된 혈청 영양소 수준을 조사한 결과 혈청 엽산, 비타민 B₁₂, 25(OH)D₃, 철, 페리틴의 수치는 대부분 정상 범위에 해당하며 두 군 간의 유의적인 차이는 없었다. 본 연구에서 비만과 고도비만 성인 여성의 혈청 영양소 수준은 유의적 차이가 없는 것으로 나타났다. 국내에서 비만 예방이나 치료를 위한 근거를 마련하기 위해서 비만과 영양소의 관련성에 대해 보다 심도 깊은 전향적 연구가 필요할 것으로 사료된다.

• Advances in Traditional Medicine
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• 제9권3호
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• pp.259-267
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• 2009

Curcumin (CMN) is known to have beneficial role in anorexia, coryza, cough, diabetic wounds, and hepatic disorders apart from its inherent antioxidant effects. Therefore, the present study was aimed to evaluate antioxidant effect of CMN in prevention of nephrolithiasis in rats-induced by ethylene glycol (EG) and Vitamin D3 (Vit. D3). Male Wistar rats (175 - 200 g) were randomized in groups like control, EG + Vit. D3 induced nephrolithiatiatic rats, CMN treated rats, CMN + EG + Vit. D3 treated rats, Vit. E + EG + Vit. D3 treated rats. Urine was collected weekly throughout the experimental protocol and estimated for calcium oxalate (CaO) count. After completion of experimental protocol serum was estimated for blood urea nitrogen and creatinine. Both the kidneys were excised and used to evaluate levels of biomarkers of oxidative stress and calcium oxalate crystal deposition by histopathological studies. Administration of EG and Vit. D3 to rats resulted in increased oxidative stress, hyperoxaluria and renal deposition of CaO crystals. Supplementation with CMN improves kidney function, reduces elevated oxidative stress, urinary oxalate level and renal deposition of CaO which shows its protective action in nephrolithiasis. The increased deposition of stone in the kidney and stone forming constituents of nephrolithiatic rats were effectively lowered by treatment of CMN.

• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.261-267
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• 2002

The aim of the present study was to investigate the interaction of $estradiol-17{\beta}-bovine$ serum albumin $(E_2-BSA)$ and calcitropic hormones, such as parathyroid hormone, calcitonin, and vitamin D, in regulation of $Ca^{2+}$ uptake in primary cultured renal proximal tubule cells. Statistically significant increase in $Ca^{2+}$ uptake was found from 2 hours after $(E_2-BSA)\;(10^{-9}\;M)$ treatment, while $estradiol-17{\beta}\;(10^{-9}\;M)$ did not affect. Treatment of the cells with $(E_2-BSA)\;(10^{-9}\;M)$ together with parathyroid hormone (PTH) $(10^{-8}\;M),$ vitamin D $(10^{-8}\;M),$ or calcitonin $(10^{-8}\;M)$ significantly stimulated $Ca^{2+}$ uptake by 32.50%, 29.30%, or 27.75%, respectively, compared with the control. However, calcitropic hormones did not exhibit any synergistic effect on the E2-BSA-induced stimulation. $E_2-BSA$ significantly increased cAMP generation and PKC activity. The stimulatory effect of cotreatment of $E_2-BSA$ and PTH or vitamin D was blocked by SQ22536 (an adenylate cyclase inhibitor) and staurosporine (a PKC inhibitor), but the effect of cotreatment of $E_2-BSA$ and calcitonin was not blocked. Furthermore, 8-Br-cAMP and TPA (an artificial PKC promoter) increased $Ca^{2+}$ uptake by 25.51% and 16.47%, respectively, compared with the control. In conclusion, $E_2-BSA$ combined with calcitropic hormones regulated $Ca^{2+}$ uptake partially via cAMP and PKC-dependent mechanisms in renal proximal tubule cells.

• Journal of Nutrition and Health
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• 제47권5호
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• pp.321-329
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• 2014

제5기국민건강영양조사 자료중 40세이상 남성을 대상으로 혈액 중 25(OH)D와 COPD의 상관관계를 조사하였다. 25(OH)D는 사분위로 군을 나누어 적용하였고, 2012년 개정된 진료지침에 따라 $FEV_1/FEV_6$ < 0.73인 경우를 COPD로 판정하였다. 다른 요인을 보정하지 않았을 경우에는 25(OH)D와 COPD 사이에 상관관계가 없었으나 흡연상태, 가계소득, 교육수준, 직업, BMI, 만연령및흡연지수를 보정하였을 때는 25(OH)D와 COPD사이에 유의한 상관관계를 보였다. 25(OH)D농도 17~21 ng/mL인 3/4분위군을 기준으로 1/4분위의 경우의 OR 1.643 (95% CI 1.161-2.236), 2/4분위는 OR 1.453 (95% CI 1.045-2.020)으로 유의하게 증가하였다. 결론적으로 40세 이상 남성에서 혈중 25(OH)D 농도는 COPD 유병가능성과 유의한 상관관계를 보였다. 즉, 3/4 분위군 [25(OH)D 17 ng/mL 이상 21 ng/mL 미만]을 기준으로 혈중농도가 낮을수록 COPD 유병가능성이 높았다.

• Tuberculosis and Respiratory Diseases
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• 제77권2호
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• pp.73-80
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• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

• 한국식품영양과학회지
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• 제13권1호
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• pp.27-32
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• 1984

최근(最近) 산업발달(産業發達)과 더불어 산업폐기물(産業廢棄物)중의 유해(有害) 중금속(重 金屬)에 의한 중독(中毒) 사례(事例)는 날로 증가(增加)하고 있는 실정(寶情) 이다. 그 중 금만성중독시(急慢性中毒時) 심각한 병적(病的) 증상(症狀)을 일으키는 Cd과 다른 영양소(營姜素)와의 상호작용(相互作用)을 규명(究明)코저 한 하나의 시도(試圖)로서 저자(善者)는 다량(多量) 무기영양소(無機營養素)중의 중요(中要) 성분(成分)인 Ca과 Ca의 흡수(吸收)에 중요(重要)한 영향(影響)을 미치는 vitamin D가 Cd 중독(中毒)에 미치는 영향(影響)을 조사(調査)하기 위하여 $CdCl_2$용액(溶液)을 음료수(Cd 로서 20 ppm)로 하고 Ca 및 Ca과 vitamin D를 첨가(添加)한 식이(食餌)를 Sprague-Dawley 계(系) 흰쥐(female)에게 80일간(日間) 급여(給與) 시켰던 바 다음과 같은 결과(結果)를 얻었다. l. 체중증가량(體重增加量)은 Cd 급여(給與)로 현저히 감소(減少)하였으나 Ca과 vitamin D 동시(同時) 급여(給與)로 제어(抑制)되었으며 각종(各種) 장기중량(臟器重量)에서 Cd급여(給與)로 인하여 신장(腎臟)과 비장(脾臟)의 중량(重量)은 유의(有意)하게 증가(壇加)하였으나 Ca 및 Ca과 vitamin D의 동시(同時) 급여(給與)로 제어(抑制)되었다. 2. 혈액(血液)중의 Hematocrit 치(値)는 Cd급여(給興)로 현저히 감소(減少)하였으나 Ca 및 Ca과 vitamin D의 동시급여(同時結與)로 제어(抑制)되었으며 cholesterol은 Cd급여(絡與)한 모든 군(群)에서 증가(增加)하였고, GOT, GPT, 종단백질(總蛋白質), albumin, globu1in 및 A/G ratio는 유의성(有意性)이 나타나지 않았다. 3. 간장(肝臟)중의 무기질(無機質) 함량(含量)은 Cd급여(給與)로 인하여 증가(增加) 하였던 Cd과 Mg이 Ca 및 Ca과 vitamin D의 동시(同時) 급여(給與)로 감소(減少)하였고 Cu는 전체적(金體的)으로 변화(變化)가 없었으며 Zn은 Cd 급여(給與)로 감소(減少)하였으나 Ca과 vitamin D의 급여(給與)에 의하여 증가(增加)하였고 Ca은 Ca과 viamin D의 급여(給輿)로 유의(有意)하게 증가(增加)하였다. 신장(腎臟)중의 무기질(無機質) 함량(含量)은 Cd급여(給輿)로 Cu, Mg은 감소(滅少)하였으나 Ca, Zn은 변화(變化)가 없었고 Ca 및 Ca과 Vitamin D의 급여(給與)로 Cd, CU, Zn은 증가(增加)하였다.

• 대한약침학회지
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• 제9권2호
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• pp.17-37
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• 2006

Objectives : To observe the changes in the serum proteins after intravenous injection of cultivated wild ginseng pharmacopuncture. Methods : Blood was collected before and after the administration of cultivated wild ginseng pharmacopuncture and only the serum was taken. Then differences in the spots on the scanned image after carrying out 2-Dimensional electrophoresis were located and conducted mass analysis and protein identification. Results : Following results were obtained from the comparative analysis of serum proteins before and after the administration of cultivated wild ginseng pharmacopuncture. 1. 28 spots were identified before and after the administration. 2. In confirming manifestation degree, spots with more than two-times increase were 204, 1302, 2205, 3105, 7104, 8006, spots with more than one-time increase were 1101, 1505, 2013, 2403, 3009, 3010, 4002, 4009, 6704, 8101, and spots with decrease were 205, 801, 803, 3205, 5202, 6105, 6106, 7103, 9001, 9003. 3. After conducting protein identification, proteins 205, 804, 1302, 4009, 6105, 6106 are unidentified yet, and 1l01 is unnamed protein. Protein 204 is identified as complement receptor CR2-C3d, 801 as YAPl protein, 803 as antitrypsin polymer, 1505 as PRO0684, 2013 and 3010 as proapolipoprotein, 2205 as USP48, 2403 as vitamin D binding protein, 3009 as complement component 4A preprotein, 3105 as immunoglobulin lambda chain, 3205 as transthyretin, 4002 as Ras-related protein Ral-A, 4204 as beta actin, 5202 and 7104 as apolipoprotein Ll, 6704 as alpha 2 macroglobulin precursor, 7103 as complement component 3 precursor, 8006 as testis-specific protein Y, 8101 as transferrin, 9001 as (Alpha-Oxy, Beta-(Cl12g)deoxy) T-State Human Hemoglobin, and 9003 as human hemoglobin. 4. Immune protein CR2-C3d(204), which acts against microbes and pathogenic organisms, was increased by more than two-times after the administration of pharmacopuncture. 5. Antitrypsin(803), which is secreted with inflammatory response in the lungs, was reduced after the administration of pharmacopuncture. 6. Proapolipoprotein(2013, 3010) and apolipoprotein(7104), key components of the HDL-cholesterol which plays an important role in preventing arteriosclerosis, were increased after the administration of pharmacopuncture. 7. Vitamin D binding protein(DBP, 2403), protecting the lung at the time of inflammatory response, was increased after the administration of pharmacopuncture. 8. Transthyretin(TTR, 3205), which is the main protein causing familial amyloid polyneuropathy(FAP), was decreased after the administration of pharmacopuncture. 9. Ras-related protein Ral-A(4002) that controls phospholipid metabolism, cytoskeletal formation, and membrane traffic, was increased after the administration of pharmacopuncture. 10. Testis-specific protein Y(8006), which takes part in determination of the gender, was increased by more than two-times after the administration of pharmacopuncture. 11. Transferrin(8101), which balances the iron level in the body, was increased after the administration of pharmacopuncture. Conclusion : Above results support the notion that intravenous injection of cultivated wild ginseng pharmacopuncture induce changes in serum proteins and this research can be a pioneer work in finding biomarkers.

• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.865-873
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• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.