• Clinical and Experimental Reproductive Medicine
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• 제29권2호
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• pp.117-127
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• 2002

Objective: This study was to investigate the generation of the functional neuron derived from human embryonic stem (hES, MB03) cells on in vitro neural cell differentiation system. Methods: For neural progenitor cell formation derived from hES cells, we produced embryoid bodies (EB: for 5 days, without mitogen) from hES cells and then neurospheres (for $7{\sim}10$ days, 20 ng/ml of bFGF added N2 medium) from EB. And then finally for the differentiation into mature neuron, neural progenitor cells were cultured in i) N2 medium only (without bFGF), ii) N2 supplemented with 20 ng/ml platelet derived growth factor-bb (PDGF-bb) or iii) N2 supplemented with 5 ng/ml brain derived neurotrophic factor (BDNF) for 2 weeks. Identification of neural cell differentiation was carried out by immunocytochemistry using $\beta_{III}$-tubulin (1:250), MAP-2 (1:100) and GFAP (1:500). Also, generation of functional neuron was identified using anti-glutamate (Sigma, 1:1000), anti-GABA (Sigma, 1:1000), anti-serotonin (Sigma, 1:1000) and anti-tyrosine hydroxylase (Sigma, 1:1000). Results: In vitro neural cell differentiation, neurotrophic factors (PDGF and BDNF) treated cell groups were high expressed MAP-2 and GFAP than non-treated cell group. The highest expression pattern of MAP-2 and $\beta_{III}$-tubulin was indicated in BDNF treated group. Also, in the presence of PDGF-bb or BDNF, most of the neural cells derived from hES cells were differentiated into glutamate and GABA neuron in vitro. Furthermore, we confirmed that there were a few serotonin and tyrosine hydroxylase positive neuron in the same culture environment. Conclusion: This results suggested that the generation of functional neuron derived from hES cells was increased by addition of neurotrophic factors such as PDGF-bb or BDNF in b-FGF induced neural cell differentiation system and especially glutamate and GABA neurons were mainly produced in the system.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.122-127
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• 2023

심혈관 질환(CVD)은 전 세계적으로 주요 사망 원인으로써 갈수록 증가하는 추세이며, 혈관 손상이 발생하였을 때, 혈전이 과도하게 형성되는 것이 그 원인인 중 하나이다. 근래에 혈소판 억제를 통한 항혈전 물질에 대한 관심이 커지고 있으며 천연 생물활성 화합물을 사용함으로써 부작용을 줄이려는 노력이 이루어지고 있다. Flavonoid 중 하나로 알려진 hydroxygenkwanin(HGK)은 팥꽃나무(Daphne genkwa)에서 정제되는 물질로서 항균, 항염증 및 항암 효과가 있다고 알려져 있으며, 혈전증을 예방하는 조직 인자의 억제제 역할을 한다고 보고되었지만 항혈소판 효과와 그 작용기전에 대해서는 거의 알려지지 않았다. 본 연구를 통해 HGK가 collagen 유도의 사람 혈소판 응집에 미치는지 확인하였고, 그 작용 기전을 확인하였다. HGK은 혈소판 신호 전달 과정에서 PI3K/AKT 및 MAPK의 인산화를 억제하였고, ATP 및 serotonin 등의 혈소판 내 과립 분비를 감소하였다. . 또한, HGK는 cPLA₂의 인산화를 억제하며 응집 촉진물질인 TXA₂ 생성을 강하게 저해하였다. 결과적으로 응집 유도 물질인 collagen가 유도한 혈소판 응집을 86.36 µM의 IC₅₀로 강하게 억제하였다. 그러므로, 본 연구를 통해 HGK가 혈관 손상을 통해 일어나는 사람 혈소판의 활성화 및 응집을 억제하는 항혈전 물질로 가치가 있음을 분명히 하였다.

• 대한한방내과학회지
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• 제18권1호
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• pp.69-85
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• 1997

This study was designed to elucidate the anti-inflammatory, cardiovascular, anti-thrombotic and analgesic effects of Shintongchugeotang. The anti-inflammatory effect was measured by the method of carragenin induced edema, protein leakage test using CMC-pouch, and the analgesic effect was measured by the acetic acid method and hot plate method, and the effect of Shintongchugeotang on the cardiovascular system was observed by the change of flow rate of Ringer solution in the vascular system in the ear of rabbit, and the contraction and dilatation of rat tail artery. Death rate, platelet aggregation, plasma coagulation activity was observed for the measurement of the anti-coagurative effect of Shintongchugeotang. The result was as follows : 1. After the administration of Shintongchugeotang extract, Carragenin induced edema and CMC-pouch protein leakage were significantly decreased. 2. The slight analgesic effect of Shintongchugeotang extract was confirmed by the observation of writhing syndrome, paw licking time, and escape time. 3. The drug increased the auricular blood flow in rabbit. 4. The drug relaxed the artery contraction by pretreated norepinephrine in rat. 5. The drug inhibited the death rate of mouse which was led to thromboembolism by serotonin and collagen. 6. The drug inhibited the platelet aggregation in rat. 7. The drug prolonged the prothrombin time and activated partial thromboplastin time on the test of plasma coagulation factor activity in rat, but was not valuable.

• Journal of Ginseng Research
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• 제47권5호
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• pp.638-644
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• 2023

Background: The anti-platelet activity of the saponin fraction of Korean Red Ginseng has been widely studied. The saponin fraction consists of the panaxadiol fraction (PDF) and panaxatriol fraction (PTF); however, their anti-platelet activity is yet to be compared. Our study aimed to investigate the potency of anti-platelet activity of PDF and PTF and to elucidate how well they retain their anti-platelet activity via different administration routes. Methods: For ex vivo studies, Sprague-Dawley rats were orally administered 250 mg/kg PDF and PTF for 7 consecutive days before blood collection via cardiac puncture. Platelet aggregation was conducted after isolation of the washed platelets. For in vitro studies, washed platelets were obtained from Sprague-Dawley rats. Collagen and adenosine diphosphate (ADP) were used to induce platelet aggregation. Collagen was used as an agonist for assaying adenosine triphosphate release, thromboxane B2, serotonin, cyclic adenosine monophosphate, and cyclic guanosine monophosphate (cGMP) release. Results: When treated ex vivo, PDF not only inhibited ADP and collagen-induced platelet aggregation, but also upregulated cGMP levels and reduced platelet adhesion to fibronectin. Furthermore, it also inhibited Akt phosphorylation induced by collagen treatment. Panaxadiol fraction did not exert any antiplatelet activity in vitro, whereas PTF exhibited potent anti-platelet activity, inhibiting ADP, collagen, and thrombin-induced platelet aggregation, but significantly elevated levels of cGMP. Conclusion: Our study showed that in vitro and ex vivo PDF and PTF treatments exhibited different potency levels, indicating possible metabolic conversions of ginsenosides, which altered the content of ginsenosides capable of preventing platelet aggregation.

• The Korean Journal of Pain
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• 제37권1호
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• pp.26-33
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• 2024

Background: Sitagliptin is an antidiabetic drug that inhibits dipeptidyl peptidase-4 enzyme. This study aimed to investigate the antinociceptive and anti-inflammatory effects of sitagliptin in formalin and carrageenan tests and determine the possible mechanism(s) of its antinociceptive activity. Methods: Male Swiss mice (25-30 g) and male Wistar rats (180-220 g) were used for formalin and carrageenan tests, respectively. In the formalin test, paw licking time and in the carrageenan test, paw thickness were considered as indexes of pain behavior and inflammation respectively. Three doses of sitagliptin (2.5, 5, and 10 mg/kg) were used in these tests. Also, several antagonists and enzyme inhibitors were used to evaluate the role of adrenergic, serotonergic, dopaminergic, and opioid receptors as well as the NO/cGMP/K_ATP pathway in the antinociceptive effect of sitagliptin (5 mg/kg). Results: Sitagliptin showed significant antinociceptive and anti-inflammatory effects in the formalin and carrageenan tests respectively. In the carrageenan test, all three doses of sitagliptin significantly (P < 0.001) reduced paw thickness. Pretreatment with yohimbine, prazosin, propranolol, naloxone, and cyproheptadine could not reverse the antinociceptive effect of sitagliptin (5 mg/Kg), which indicates that adrenergic, opioid, and serotonin receptors (5HT₂) are not involved in the antinociceptive effects. L-NAME, methylene blue, glibenclamide, ondansetron, and sulpiride were able to reverse this effect. Conclusions: NO/cGMP/K_ATP, 5HT₃ and D₂ pathways play an important role in the antinociceptive effect of sitagliptin. Additionally significant anti-inflammatory effects observed in the carrageenan test might contribute in reduction of pain response in the second phase of the formalin test.

• 대한한방소아과학회지
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• 제12권1호
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• pp.231-256
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• 1998

Experimental studies were done to research the clinical effects of Chihyosan and Chihyosangamibang on the Anti-allergic effect and pulmonary function of $O_3$ intoxicated Rats. Anti-allergic effect experiment consisted of vascular permeability responses to intradermal histamine and serotonin, 48hrs homologous passive cutaneous anaphylaxis provoked by the IgE-like antibody against egg white albumin, and delayed type hypersensitivity responses to Picryl Chloride and SRBC. Pulmonary function of $O_3$ intoxicated Rats experiment consisted of lung TBA value, water Contents of the lung, oxygen consumption time, and arterial blood $pCO_2,\;pO_2,\;HCO_3^-$, pH level. The results obtained as follows; 1. In the effects of Chihyosan and Chihyosangamibang on vascular permeability responses to intradermal histamine, both of chihyosan and Chihyosangamibang group revealed significant effect. 2. In the effects of Chihyosan and Chihyosangamibang on vascular permeability responses to intradermal serotonin, both of chihyosan and Chihyosangamibang group revealed significant effect. 3. In the 48hrs homologous passive cutaneous anaphylaxis provoked by the IgE-like antibody against egg white albumin, Chihyosan groups revealed significant effect, but Chihyosangamibang groups revealed none significant effect. 4. In the delayed type hypersensitivity responses to Picryl Chloride, Chihyosan and Chihyosangamibang groups revealed none significant effect. 5. In the delayed type hypersensitivity responses to. SRBC, Chihyosan revealed none significant effect, but Chihyosankamibang revealed significant effect. 6. Both of Chihyosan and Chihyosangamibang groups revealed significant effect on decrease of the lung TBA value of lung. 7. Both of Chihyosan and Chihyosangamibang groups revealed significant effect on decrease of the water contents of right and left lung. 8. Both of Chihyosan and Chihyosangamibang groups revealed significant effect on decrease of oxygen consumption time. 9. In the decrease effect of arterial blood $pCO_2$ level, both of Chihyosan and Chihyosangamibang groups revealed none significant effect. 10. In the increase effect of arterial blood $pO_2$ level, both of Chihyosan and Chihyosangamibang groups revealed none significant effect. 1. In the decrease effect of arterial blood $HCO_3^-$ level, both of Chihyosan and Chihyosangamibang groups revealed significant effect. 12. In the increase of arterial blood pH level, Chihyosangamibang groups revealed none significant effect, but Chihyosan groups revealed significant effect. According to above stated results, both of Chihyosan and Chihyosangamibang are very usefully for treatment of cough, asthma, chronic obstructive pulmonary diseases and allergic pulmonary diseases.

• 대한한방소아과학회지
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• 제8권1호
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• pp.27-37
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• 1994

This Experimental study was done to investigate the effect of the Taklisodok-um on the Anti-allergic response. The results were obtained as follows: 1. On vascular permeability response to the intradermal injected Histamine, the Taklisodok-um treated group revealed more significant decrease than control group. 2. On vascular permeability response to the intradermal injected Serotonin, the Taklisodok-um treated group revealed more significant decrease than control group. 3. In the 48hrs homologous passive cutaneous anaphylaxis provoked by the IgE-like antibody against white egg albumin, there was no significant difference between the Taklisodok-um treated group and control group. 4. The Taklisodok-um treated group revealed more significant inhibitory effect than control group in the delayed type hypersensitivity response to Picryl chloride. 5. The Taklisodok-um treated group revealed more significant inhibitory effect than control group in the delayed type hypersensitivity response to SRBC. According to the above-stated results, it is considered that the Taklisodok-um could be applied widely to the type I and IV allergic diseases.

• 대한한방소아과학회지
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• 제8권1호
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• pp.13-26
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• 1994

Experimental studies were done to research the clinical effects of Sipyungtang and Sipyungtang-gamibang (sipyungtang with Anemarrhenae Rhizoma added) on the Anti-allergic effect. The results obtained as follows; 1. In the effects of Sipyungtang and Sipyungtang-gamibang on vascular permeability responses to intradermal Histamine, Sipyungtang group revealed none significant effect, but Sipyungtang-gamibang group revealed significant effect. 2. In the effects of sipyungtang and Sipyungtang-gamibang on vascular permeability responses to intradermal serotonin, Sipyungtang group revealed none significant effect, but Sipyungtang-gamibang group revealed significant effect. 3. In the 48hrs homologous passive cutaneous anaphylaxis provoked by the IgE-like antibody against egg white albumin, Sipyungtang and Sipyungtang-gamibang groups revealed none significant effect. 4. In the delayed type hypersensitivity responses to Picryl Chloride, Sipyungtang and Sipyungtang-gamibang groups revealed none significant effect. 5. In the delayed type hypersensitivity responses to SRBC, Sipyung-tang and Sipyungtang-gamibang groups revealed significant effect.

• 대한한방소아과학회지
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• 제6권1호
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• pp.1-14
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• 1992

Experimental studies were done to research the clinical effects of Juguieum and Juguieum-gamibang(Juguieum with Lonicerae Flos and Forsythiae Fructus added) on the Antiallergic effect. The results obtained as follows; 1. In the effects of Jugieum and Jugieum-gamibang on vascular permeability responses to intradermal histamine, Juguieum group revealed none significant effect, but Jugieum-gamibang group revealed significant effect. 2. In the effects of Juguieum and Juguieum-gamibang on vascular permeability responses to intradermal serotonin, Juguieum and Juguieum-gamibang groups revealed significant effect. 3. In the 48hrs homologous passive cutaneous amaphylaxis provoked by the IgE-like antibody against egg albumin, Juguieum and Juguieum-gamibang groups revealed significant effect. 4. In the delayed type hypersensitivity responses to Picryl Chloride, Juguieum and Juguieum-gamibang groups revealed significant effect. 5. In the delayed type hypersensitivity responses to SRBC, Juguieum group revealed significant effect, but Jugieum-gamibang group revealed none significant effect. According to above-stated results, Juguieum and Juguieum-gamibang are concluded to be effective as anti-allergic regimen and recommended to be used for the treatment of type I and type IV allergic disease.

• 한국동물위생학회지
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• 제46권2호
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• pp.107-113
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• 2023

Tenebrio molitor, rich in protein, is used as an alternative protein source. Many studies have evaluated Tenebrio molitor larvae (TML) for anti-obesity effects, fatty liver relief, antithrombosis, and antioxidant effects. In this study, we prepared an experimental diet by adding Tenebrio molitor larvae oil (TMLO) to feed and administered it to adult male ICR mice for six weeks. The study assessed the weight gain of mice, prothrombin time (PT), activated partial thromboplastin time (aPTT), blood coagulation time, and the activities of coagulation factor VII and coagulation factor XII. The weight gain of mice was suppressed in the groups fed with TMLO, indicating a potential anti-obesity effect. The blood coagulation time was delayed in the TMLO-fed groups, as evidenced by amplified PT and aPTT values. Furthermore, the activities of coagulation factor VII and coagulation factor XII were diminished in the TMLO-treated groups, indicating a potential thrombosis relieving effect. The concentrations of thromboxane B2 and serotonin declined in the TMLO-fed groups, signifying potential blood circulation effects. Finally, blood triglyceride and LDL-cholesterol concentrations were reduced in the TMLO-fed groups. In summary, TMLO supplementation may have an inhibitory effect on liver damage and blood clot formation caused by obesity.