• The Korean Journal of Physiology
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• 제23권1호
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• pp.13-21
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• 1989

The present study was designed to investigate i) the action of various nucleotides on membrane permeability of rat red blood cell and hepatocyte for $Na^{+}$ and $Rb^{+}$ ii) the characteristics of purinoceptors on these cell membranes. Blood from Sprague-Dawley rats was obtained by carotid arterial cannulation. Red blood cells were then washed 3 times with saline at $4{\circ}C$. Hepatic parenchymal cells were isolated from rat livers by using a modification of the Berry and Friend (1969) method. For the $Na^{+}$ influx studies, isolated RBC and hepatocyte were incubated in incubation medium containing $^{22}Na^{+}0.2\;{\mu}Ci/ml$ at $37^{\circ}C$. After various time intervals samples were removed from the incubation flask and washed out 3 times with ice-cold washing solutions. Cells were destroyed by adding Triton X-100 and TCA solution. After centrifugation, the supernatants were assayed for $^{22}Na^{+}$ by gamma counter. $^{86}Rb^{+}$ was used to simulate $K^{+}$ in these $K^{+}efflux$ studies. Isolated hepatocytes were incubated for 60 min in the loading solution containing $^{86}Rb^{+}\;10\;{\mu}Ci/ml$ at $37^{\circ}C$. After loading, the cells washed out 3 times by centrifugation with washing solution. The cells were incubated in buffer solution at $37^{\circ}C$. At intervals thereafter, samples were removed and centrifuged. The supernatants were analyzed for $^{86}Rb^{+}$ by liquid scintillation counter. The main results of the experiments were: 1) ATP and ATPP increased in both $^{22}Na^{+}$ influx and $^{86}Rb^{+}$ efflux in the red blood cell. Although ADP showed a tendency to increase in RBC membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$, the changes were not significantly different from the control. 2) The Significant changes in $^{22}Na^{+}$ and $^{86}Rb^{+}$ flux by ATP were also demonstrated in hepatocyte. ATPP and ADP showed a tendency to increase in hepatocyte membrane permeability for both ions. 3) Other nucleoside triphosphates-ITP, GTP and CTP-did not change in membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$ in RBC and hepatocyte. In conclusion, not only ATP but also ATPP activate purinoceptors and change in membrane permeability for $Na^{+}$ and $K^{+}$. In order to activate purinoceptors on the cell membrane, the nucleotides have to possess intact adenine moiety and three phosphates or more in its molecule.

• 생약학회지
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• 제27권2호
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• pp.111-116
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• 1996

The components were isolated from the root of Astragalus membranaceus and their structures were characterized as 3,4-methylenedioxypyrrolealdehyde, $7-O-{\beta}-D-glucopyranosyl$ 7, 3'-dihydroxy-4'-methoxy isoflavone and a naphthalene derivative on the basis of spectral and physical methods. $7-O-{\beta}-D-glucopyranosyl$ 7, 3'-dihydroxy-4'-methoxy isoflavone and the naphthalene compound showed protective effect on $CC_{l4}-induced$ cytotoxicity in primary cultured rat hepatocytes.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
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• pp.58-64
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• 1990

The effects of ginseng saponins, G-Rbl and G-Rc on the rat liver LDH A-gene transcnptional activity was investigated during pro-replicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl of G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 ma/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5·hours after partial hepatectomy. Dose dependent effect of G-Rbl and G-Rc (1-25 mg/100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal in- creases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc However, when the administration doses of G-Kbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rb 1 and G·Rc have stimulatory effect at the lower concentration (1 mg/100g B.W) and inhibitory effect at the higher concentration (20 moi loos 5.W) on the LDH A-gene transcription during regeneration of rat liver, Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA synthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G·Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 ug and 100 ug/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen enhancer activity for the hepatocyte proliferation during rat liver regeneration period. Keywords Inductive effects of ginsenosides, G-Rb, -Rc, and -Re, rat LDH A-gene transcription, the sin thetic rate of proteins and DNA in regeneration rat liver.

• Journal of Ginseng Research
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• 제14권2호
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• pp.200-206
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• 1990

The effects of ginseng saponins, SRbl and G-Rc on the rat liver LDH A-gene transcriptional activity was investigated during prereplicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl or 'G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 mg/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5-hours after partial hepatectomy Dose dependent elect of G-Rbl and G-Rc (1-25 mg/ 100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal increases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc. However, when the administration doses of G-Rbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver. In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rbl and G-Rc have stimulatory effect at the lower concentration (1 mg/ 100g B.W) and inhibitory effect at the higher concentration (20 mg/ 100g B.W) on the LDH A-gene transcription during regeneration of rat liver. Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA sinthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G-Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 us and 100 $\mu\textrm{g}$/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen ehincer activity for the hepatocyte proliferation during rat liver regeneration period.

• 한국수산과학회지
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• 제29권2호
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• pp.246-255
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• 1996

연속식 3단계 막반응기로부터 각 단계별로 분리한 가자미피 젤라틴 가수분해물의 항산화성을 측정한 결과, 2단계 가수분해물의 항산화력이 가장 뛰어날 뿐만 아니라, 천연항산화제인 $\alpha-tocopherol$보다 $10\%$ 정도의 높은 항산화력을 나타낸 반면, 1단계 및 3단계 가수분해물은 $\alpha-tocopherol$보다 오히려 $10\~15\%$ 정도 낮은 항산화력을 보였다. 가수분해물의 첨가한 농도에 따른 항산화성은 유지중량에 대해 $1.0\%(w/w)$로 첨가한 농도에서 최대 항산화력을 나타내었다. 한편, 천연항산화제인 $\alpha-tocopherol$과 합성항산화제인 BHT와의 상승효과를 검토한 결과, 각 단계별 가수분해물은 $\alpha-tocopherol$과 우수한 상승효과를 관찰할 수 있었으며, 그 중 2단계 가수분해물의 상승효과가 가장 강하였다. 또한 항산화성이 가장 뛰어난 2단계 가수분해물로부터 gel column, ion exchange column 및 ODS column을 사용하여 항산화력이 특히 우수한 부분만을 분리한 단일 펩티드를 간세포에 첨가하여 세포생존율에 미치는 효과를 관찰한 결과, TBHP의 독성에 대해 펩티드 첨가구가 무첨가구에 비해 세포생존을 연장시켰으며, lipid peroxidation측정에서도 세포의 산화를 억제함으로써 세포의 생존율을 높였다.

• Applied Biological Chemistry
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• 제40권6호
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• pp.577-582
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• 1997

감귤류에 포함되어있는 성분중, 생리활성물질로 거론되어지고 있는 citrus flavonoid 즉, hesperidin, hesperetin, naringin 및 naringenin 4종류를 가지고 간장triglyceride합성의 율속효소로 알려져 있는 phosphatidate phosphohydrolase 활성에 대하여 in vitro계에 미치는 영향과 이들 flavonoid가 HepG2 세포의 생존율 및 증식에 미치는 영향에 대해서도 함께 검토하였다. 그 결과 본 실험에 사용된 모든 flavonoid 는 $10{-4}M$ 농도까지는 세포생존율 및 증식이 최고에 도달하였으나, $10{-3}M$ 농도에서 aglycone flavonoid는 현저한 저해효과를 나타냈다. naringin 및 hesperidin의 glycoside flavonoid는 첨가한 모든 농도에서 PAP활성을 촉진시키는 경향을 나타내었다. 그러나, aglycone flavonoid인 hesperetin은 PAP활성을 농도 의존적으로 저해하였으며, naringenin은 $10{-2}M$ 농도에서 강한 저해효과를 나타내었다. 이러한 결과로 미루어 볼 때 aglycone flavonoid가 생체내에서 세포 생존율, 증식 및 지질대사에 강한 영향을 미칠 것으로 생각되어진다.

• Toxicological Research
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• 제25권1호
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• pp.29-33
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• 2009

Phellinus gilvus (PG) is a widely used mushroom for health promotion. We studied the hepatoprotective effect of the polysaccharide aqueous extract of PG (PGP) against carbon tetrachloride ($CCl_4$)-induced liver injury in rats. Sprague Dawley rats were divided into 5 groups: Normal control, $CCl_4$ control, PGP 50, 100, and 200 mg/kg + $CCl_4$. The levels of serum biochemical parameters, liver lipid peroxide and antioxidant enzymes, and histological appearances were evaluated. The $CCl_4$-induced increments of alanine aminotransferase, asparate aminotransferase, and alkaline phosphatase levels in serum were significantly decreased by PGP-pretreatments. The PGP dose-dependently decreased hepatic malondialdehyde formations in $CCl_4$-treatment groups. Hepatic antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) were elevated by PGP in $CCl_4$-treatment groups. Histopathological evaluation of liver showed that the loss of hepatocytes, fatty changes, swelling and extensive necrosis of hepatocytes in centrilobular regions of the $CCl_4$-treated rats were ameliorated by PGP pretreatment. The PGP has hepatoprotective and antioxidative effects in $CCl_4$-induced liver injury of rat.

• 한국식품영양과학회지
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• 제40권1호
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• pp.78-83
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• 2011

최근 국내의 건강기능성 추출물을 이용한 차 음료 시장이 확대되고 있고, 국화, 산수유 및 오미자는 이들의 높은 생리 활성과 기호도로 인하여 그 활용도가 더욱 높을 것으로 기대된다. 현재까지 국화, 산수유 및 오미자의 개별적인 생리활성 연구는 활발히 이루어지고 있는 반면, 이들을 혼합하여 제조한 차류의 효능에 대한 연구는 아직 보고되지 않고 있는 실정이다. 따라서 본 연구에서는 $CCl_4$ 처리에 의해 손상된 간세포에 대한 국화차 조성물의 간 기능 보호효과와 인체백혈구에서 $H_2O_2$로 유도된 산화적 DNA 손상에 대한 국화차 조성물의 항유전독성 효과를 검증하고자 하였다. $CCl_4$로 유도된 간세포 독성에 대해 국화차 조성물 I형, II형은 유의적인 간세포 보호효과를 나타내었으며, 국화차 조성물 III형은 $CCl_4$처리군보다 약간 높은 세포생존율을 보였으나 유의적이지는 않았다. $CCl_4$를 처리에 의해 증가한 간세포 배지 내 AST 및 ALT 활성은 국화차 조성물에 의해 배지 내 효소활성을 감소시킴으로 간세포 보호효과가 있는 것으로 사료되며, 그 효과는 AST 효소활성에 더 큰 영향을 미치는 것으로 나타났다. 특히 국화차 조성물 I형의 0.01 mg/mL 농도에서 가장 높은 AST 및 ALT 유리 억제능을 나타내었다. 이 결과로 국화추출물의 $CCl_4$ 유도에 의해 AST>ALT 순으로 유리가 증가되는 것으로 여겨진다. 국화차 조성물 I형의 $1\sim50{\mu}g$/mL, 국화차 조성물 II의 $50{\mu}g$/mL 및 국화차 조성물 III 의 $1\sim50{\mu}g$/mL 농도 범위에서 산화적 스트레스로 유도된 인체 백혈구 DNA 손상에 대한 보호효과를 나타내었다. 이러한 결과를 통하여 국화차 조성물의 간세포 손상보호효과와 산화적 스트레스에 대한 인체 백혈구 DNA 손상 보호능을 검증할 수 있었다.

• 한국약용작물학회지
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• 제8권2호
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• pp.157-164
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• 2000

활성산소에 기인하는 다양한 성인병을 예방 치료할 수 있는 안전하고 우수한 항산화 물질을 천연물로 부터 개발하기 연구의 일환으로 오배자의 주성분에 대한 항산화효과 및 간보호효과를 연구하여 다음과 같은 결과를 얻었다. 1. Ferric-Thiocyanate 법에 의하여 과산화반응 억제에 따른 항산화 활성을 측정한 결과 모두 tocopherol보다 강한 과산화반응 억제 활성을 보였으며 gallic acid methyl ester, protocatechuic acid와 syringic acid는 tocopherol 보다는 강한 활성을 보였으며 BHA와는 유사 또는 그 이상의 활성을 보였다. 특히 1, 2, 3, 4, $6-penta-O-galloyl-{\beta}-D- glucose$는 tocopherol과 BHA보다 매우 강한 과산 화반응억제 활성을 보였다. 2. TBA 법에 의한 과산화지질의 생성의 지표인 MDA 억제효과에 의한 항산화효과를 측정한 결과 gallic acid 는 $1.6{\times}10^{-4}$의 농도에서 BHA 보다 높은 억제율을, protocatechuic acid는 $1.6{\times}10^{-6}$의 농도에서 BHA 보다 높은 억제율을 보였다. gallic acid methyl ester와 1, 2, 3, 4, $6-penta-O-galloyl-{\beta}-D-glucose$는 농도가 증가함에 따라 우수한 MDA의 억제활성을 나타내었다. 3. Glycyrrhizin을 비교군으로 하여 간 보호 실험을 한 결과 1, 2, 3, 4, $6-penta-O-galloyl-{\beta}-D-glucose$의 0.5mg/ml첨가군에서는 GOT의 강한 감소 효과를 보였으며 GPT 실험에서는 각 시료 0.5mg/ml 첨가군에서 감소하는 경향을 나타내었다.

• Archives of Pharmacal Research
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• 제18권4호
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• pp.256-261
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• 1995

The elevation of intracellular $Ca^{2+}$ in various tissue through oxidative stress induced by menadione has been well documented. Increase of $Ca^{2+}$ level inplatelets results in aggreaction of patelets. To test the hypothesis that menadione-induced $Ca^{2+}$ elevations can play a role in platelet aggregation, we have studied the effect of menadione on aggragation of platelets isolated from female rats. Treatment with menadione to platelet rich plasma (PRP), which proved to be 60% as determined by aggregometry. however, exposure of PRP to menadione leads to a loss of cell viability, as measured by lactae dehydrogenase (LDH) leakage, suggesting that menadione might induce cell lysis rather than aggregation of platelets. Turbidty changes induced by menadione were unaffected by addition ofl dicoumarol, which is a quinone reducellular factions of patelets. These data, which indicate an absence of the QR detoxifying pathway, suggest that platelets may be more susceptible to menadione-induced cytotoxicity than certain other cell, as hepatocytes.