• Title/Summary/Keyword: 5,7-DHT

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Luminescence Immunoassays and Their Applications for Dihydrotestosterone and Testosterone( I );Establishment of LIA. (Dihydrotestosterone과 Testosterone의 섬광면역 측정법과 응용( I );측정법의 정립)

  • Yoon, Yong-Dal;Kim, Sung-Rye
    • Clinical and Experimental Reproductive Medicine
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    • v.14 no.2
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    • pp.138-148
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    • 1987
  • 5${\alpha}$-dihydrotestosterone(DHT)과 testosterone(T)은 남성 생식기관의 주 생성 호르몬들로 그 구조가 매우 비슷하여 이들 각 개를 특이하게 측정(specific determination)하는 방법이 개발되지 않고 있다. 본 연구는 고속액체 크로마토그라피(HPLC)를 이용하여 이들을 분리한 후 섬광면역측 정법(Luminescence immunoassay, LIA)으로 정량하는 방법을 개발하여 이들의 응용 가능성을 검토하고져 하였다. DHT와 T의 retention time은 각각 10.3min, 17.6min이었다. DHT-LIA와 T-LIA에 서 다른 스테로이드들과의 교차반응도는 방사면역측정법(RIA)과 대동소이하였다. 정도관리(quality control) 시료의 intra-assay variation은 DHT-LIA가 8.7%, T-LIA가 6.0%의 변이계수를 나타내었고, inter-assay variation의 변이계수는 각각 12.0% 및 15.3%이었다. 실측치(y)와 기대치(x)간의 관계를 보면, DHT-LIA경우는 Y=0.94X+0.9(r=0.989), T-LIA는 Y=1.01X+0.06(r =0.988)로 두 측정치 사이에는 통계적으로 유의한 차이가 없었다. 위의-측정방법을 이용하여 DHT-enanthate와 T-enanthate 처리후 혈청내 DHT 및 T의 농도변화를 조사한 실험결과 LIA와 RIA의 값사이에 유의한 차이가 없었다. 위의 결과로 보아 본 실험에서 개발된 DHT와 T의 섬광면역측정법은 정립되었다고 사려된다.

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Concentrations of Bioavailable Testosterone and Dihydrotestosterone Determined by Luminescence Immunoassay in Serum (혈청내 섬광면역측정법에 의한 활성적 Testosterone과 Dihydrotestosterone의 농도)

  • Yoon, Yong-Dal;Lee, Chang-Joo;Chun, Eun-Hyun;Lee, Joon-Yeong
    • Clinical and Experimental Reproductive Medicine
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    • v.15 no.2
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    • pp.83-92
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    • 1988
  • 혈액에서 생물학적 활성을 나타내는 (bioavailable) steroid hormone은 주로 비결합형(free form)과 알부민 결합형(albumin-bound form)으로 구성된다. 특히 Testosterone (T)과 5 alpha-Dihydrotestosterone (DHT)의 활성적 분획이 전체의 T, DHT 양에 비해 생리적 현상과 보다 잘 일치하는 것으로 알려지고 있다. 본 연구는 섬광면역측정법(Luminescence immunoassay, LIA)으로 혈청내 활성적 T 및 DHT의 농도의 측정에 이용하고져 하였다. 항체는 T- 또는 DHT-3-CMO-BSA를 항원으로 토끼에 면역주사하여 얻었다. 추적자는 T-3-CMO, DHT-3-CMO에 aminobutylethylisoluminol(ABEI)를 부착시켜 사용하였다. 항체중 IgG분획을 Protein-A-Sepharose CL-4B로 분리한 후 Immunobead(Bio-Rad)에 부착시켜 Solid-phase LIA를 실시하였다. 본 연구에서 LIA는 정확도(accuracy), 정밀도(precision), 감도(sensitivity), 교차반응도(specificity)등을 조사하고, 기존의 방사면역측정법(RIA)과 비교하여 만족할만한 결과를 얻었다. 혈청내 T및 DHT의 활성적 분획의 농도를 측정한 결과는 다음과 같았다. T의 경우는 남성에서 T의 전체량의 33% 이상으로 $7.1{\pm}1.5nmol/l$, 여성에서는 26% 이상으로 $0.28{\pm}0.05nmol/l$이었다. DHT의 활성적 분획은 남성의 경우 $601.7{\pm}85.8pmol/l$, 여성의경우 $52.4{\pm}19.9\;pmol/l$이었다. 이상의 결과를 보아 본 연구에서 이용된 LIA는 혈청내 활성적 농도를 측정하기에 충분하다고 사료된다. 또한 이 방법을 이용하여 여성의 Androgenicity 및 남성 정소기능등의 제어방법에 응용될 수 있을 것으로 판단된다.

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Involvement of Serotonergic Mechanism in the Nucleus Tractus Solitarius for the Regulation of Blood Pressure and Heart Rate of Rats (흰쥐의 혈압 및 심박동수 조절에 대하여 Nucleus Tractus Solitarius 부위의 Serotonin성 기전의 역할)

  • Lee, Yong-Kyu;Hong, Ki-Whan;Yoon, Jae-Soon
    • The Korean Journal of Pharmacology
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    • v.25 no.1
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    • pp.1-11
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    • 1989
  • In this study, it was aimed to investigate the role of serotonergic neurotransmission in nucleus tractus solitarius (NTS) for the central regulation of blood pressure and heart rate and its involvement in baroreceptor reflex activation in rats. A microinjection of 5-hydroxytryptamine (5-HT) into the NTS produced decreases in blood pressure and heart rate. Maximal decreases were $34.4{\pm}1.6$ mmHg and $41.7{\pm}10.2$ beats per min by 300 pmol of 5-HT. Microinjections of ${\alpha}-methylnor-adrenaline$ $({\alpha}-MNE)$ and clonidine manifested similar decreases in blood pressure and heart rate. The hypotensive and bradycardial effects of 5-HT were blocked by previous applications of 5-HT antagonists, ritanserin, methysergide and ketanserin into the NTS, respectively. By pretreatment with reserpine and 6-hydroxydopamine (6-OHDA, i.c.v.), both hypotensive and bradycardial effects of 5-HT were significantly attenuated. Pretreatment with 5, 7-dihydroxytryptamine (5,7-DHT, i.c.v.) enhanced the hypotensive and bradycardial effects of 5-HT. Similarly, following pretreatment with 6-OHDA, the effects of clonidine were increased. Pretreatment either with 5,7-DHT or 6-OHDA significantly attenuated the sensitivity of baroreflex produced either by phenylephrine or by sodium nitroprusside. When either 5,7-DHT or 6-OHDA was injected into the NTS $(5,7-DHT;\;8{\mu}g\;6-OHDA;\;10{\mu}g)$, both of the baroreflex sensitivities were impaired. In the immunohistochemical study, the injection of 6-OHDA into the the NTS led to reduction of axon terminal varicosity, however, the injection did not reduce the numbers of catecholaminergic cell bodies. Likewise, when 5,7-DHT was injected into the NTS, the varicosity of serotonergic axon terminals was markedly reduced. Based on these results, it is suggested that (1) stimulation of serotonergic receptors in the NTS leads to decreases in blood pressure and heart rate as observed with the stimulation of catecholaminergic system, (2) both serotonergic and catecholaminergic receptors may be located postsynaptically, and (3) the serotonergic neurons as well as catecholaminergic neurons may have a close relevance for the activation of baroreflex.

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Microglial Reaction to the Cytotoxicity of 5,7-Dihydroxytryptamine in the Cat Retina (5,7-Dihydroxytryptamine의 세포독성에 의한 고양이 망막내 미세아교세포의 반응양상)

  • Joo, Woo-Hyun;Nam, Seong-Ahn;Jo, Seung-Mook;Cho, Hyon-Hoo;Shin, Min-Cheol;Won, Moo-Ho;Choi, Chang-Do
    • Applied Microscopy
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    • v.28 no.4
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    • pp.425-434
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    • 1998
  • This study was designed to investigate the microglial reactions to the neurodegenerative changes in the cat retina. All experiments were performed using adult cats of both sex, weighing $2,500g\sim3,500g$. 5,7-DHT $(100{\mu}g)$ dissolved in 0.1% ascorbic acid was injected into the vitreous body. All injections were performed in one-side eye; the other side served as the control, which was injected only with 0.1% ascorbic acid. Cats were sacrificed at 1, 3, 7, 14 and 21 days after intravitreal injection of 5,7-DHT For light microscopy, retinae were fixed with 4% paraformaldehyde and processed using NDPase histochemistry. Same retinae were fixed with 1% para(formaldehyde-2.5% glutaraldehyde and processed for electron microscopy. NDPase-positive microglial cells were mainly distributed in the inner plexiform layer of the retina, and characterized by a small somata with a few slender processes, which were also extended in the ganglion cell layer (GCL) and inner nuclear layer (INL). The intensity of the microglia stained for NDPase was abruptly increased at 7 day as compared with that of the control, and thereafter continuously sustained until 21 day, the last experimental group in this study. Under the electron microscopical observation, microglial cells in the control group exhibited elongate nucleus with perinuclear chromatin condensation, and the perikaryon was scanty. However, a few hypertrophic glial cells were frequently found at 3 days after the drug injection. By 7 day, most microglial cells directed toward the degenerated neurons in the GCL, and the number of microglial cells was slightly increased as compared with the former group. At the 14 day, most microglial cells wrapped the degenerated cells in the GCL, and a few cells showed phagocytotic features. By 21 day, most microglial cells were engaged in phagocytotic activity, and their cytoplasm was filled with the phagorytosed material. Based on the results, 5,7-DHT may act as a specific neurotoxin to the cat retina, and microglial reactions to the neuronal death are already induced in early experimental stage. These results indicate that the microglial cells in the cat retina show characteristic features as a protective effect of neural tissue.

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Determination of Optimal Dosage of Ethanol Extract of Houttuynia cordata Thunberg Against Benign Prostatic Hyperplasia (전립선 비대증에 대한 Houttuynia cordata Thunberg 에탄올 추출물의 최적용량 결정)

  • Lee, Jin Young;Kim, Hyung Hoi;Kang, Jae Seon
    • Journal of Life Science
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    • v.31 no.7
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    • pp.631-640
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    • 2021
  • Houttuynia cordata Thunberg has been studied for a variety of pharmacological actions in traditional oriental medicine. In this study, we investigated the effects of Houttuynia cordata ethanol extract (HCE) on benign prostatic hyperplasia (BPH) models induced by castration and testosterone propionate (TP) injection. Thirty rats were divided into six groups. One group was used as a normal control, and the other groups were castrated and had intraperitoneal injections of TP for 14 days to induce BPH. A positive control group was given daily doses of finasteride (5 mg/kg) to the BPH model. Rats administered HCE (0.5, 1 or 2 mg/kg) instead of finasteride were compared with controls as experimental groups. There was no statistical significance in terms of prostate weight based on 100 g of body weight. The concentrations of 5-α reductase and dehydroxytestosteronre (DHT) were determined via ELISA tests, and there was a significant decrease in all experimental groups. The 0.5 mg/kg HCE group had the lowest level of 5-α reductase, and the 2 mg/kg HCE group had the lowest level of DHT. In the histopathological observation of prostates, the control and the 2 mg/kg HCE groups had normal cell shapes and no swelling. However, in the negative control group and the 1 mg/kg HCE group, the cells were swollen, and the gap between the cells was narrowed. In particular, in the 0.5 mg/kg HCE group, some cells were bursting. Therefore, the administration of more than 2 mg/kg of HCE is suitable to protect against BPH.

Imbalance in the spinal serotonergic pathway induces aggravation of mechanical allodynia and microglial activation in carrageenan inflammation

  • Junxiu Jin;Dong Ho Kang;Jin Jeon;Hyung Gon Lee;Woong Mo Kim;Myung Ha Yoon;Jeong Il Choi
    • The Korean Journal of Pain
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    • v.36 no.1
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    • pp.51-59
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    • 2023
  • Background: This study investigated the effect of an excess and a deficit of spinal 5-hydroxytryptamine (5-HT) on the mechanical allodynia and neuroglia activation in a rodent pain model of carrageenan inflammation. Methods: Male Sprague-Dawley rats were implanted with an intrathecal (i.t.) catheter to administer the drug. To induce an excess or deficit of 5-HT in the spinal cord, animals were given either three i.t. 5-HT injections at 24-hour intervals or a single i.t. injection of 5,7-dihydroxytryptamine (5,7-DHT) before carrageenan inflammation. Mechanical allodynia was measured using the von Frey test for 0-4 hours (early phase) and 24-28 hours (late phase) after carrageenan injection. The changes in the activation of microglia and astrocyte were examined using immunofluorescence of the dorsal horn of the lumbar spinal cord. Results: Both an excess and a deficit of spinal 5-HT had no or a minimal effect on the intensity of mechanical allodynia during the early phase but prevented the attenuation of mechanical allodynia during the late phase, which was observed in animals not treated with i.t. 5-HT or 5,7-DHT. Animals with an excess or deficit of 5-HT showed stronger activation of microglia, but not astrocyte, during the early and late phases, than did normal animals. Conclusions: Imbalance in the descending 5-HT pathway in the spinal cord could aggravate the mechanical allodynia and enhance the activation of microglia, suggesting that the spinal 5-HT pathway plays an essential role in maintaining the nociceptive processing in balance between facilitation and inhibition in inflammatory pain caused by carrageenan inflammation.

Effect of Phellius linteus water extract on benign prostatic hyperplasia

  • Kim, Yu-Na;Kim, Min-Sun;Chun, Sung-Sik;Choi, Jeong-Hwa
    • Nutrition Research and Practice
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    • v.7 no.3
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    • pp.172-177
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    • 2013
  • Benign prostatic hyperplasia (BPH) is one of the most common diseases among elderly men. As the old-age population is increasing recently, it is to our interest to observe the growing BPH within them. In BPH, the dihydrotestosterone (DHT) acts as promotes prostate growth. It inhibits enzyme $5{\alpha}$-reductase that is involved in the conversion of testosterone to the DHT activity which reduces the excessive prostate growth. Through experiments, the effects of Phellius linteus water extract performed on the BPH rats were induced by testosterone treatments. For 12 weeks, Sprague-Dawley rats were treated with testosterone for the induction of BPH. Rats were divided into four experimental groups: the not treated group (N), the testosterone injection and D.W treatment group (TN), the testosterone injection and Phellinus linteus treatment group (TP) and testosterone injection and finasteride treatment group (TF). Prostate weight, volume and weight ratio in the TP group and the TF group were significantly lower than the TN group. Testosterone and DHT levels in the TN group were significantly higher than that of the N group. And the TP group was significantly decreased than that of the TN group. While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation; the TP and TF groups showed trophic symptoms and were lined by flattened epithelial cells, thus, the stromal proliferation is relatively low as compared to the TN group. These suggest that Phellinus linteus water extracts may be an useful remedy for treating the benign prostatic hyperplasia.

Evaluation of Antibacterial, Antioxidant Fractionalities and Hair Loss Prevention Effect of Platycodon grandiflorum (도라지 추출성분의 항균·항산화 및 탈모예방 효과)

  • Jung, Min-Hwa
    • The Korean Journal of Food And Nutrition
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    • v.32 no.5
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    • pp.553-559
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    • 2019
  • To investigate the effects of antioxidant activities and hair loss prevention of extracts from Platycodon grandiflorum, we've prepared chloroform (CF) and ethylacetate fractions (EA) extracted from P. grandiflorum. In the results of DPPH radical scavenging assay, the two fractions showed dose-dependent antioxidant activities. Furthermore, in the ABTS assay, the two fractions exhibited the inhibitory effect over 90% at 10, 50, 100, 200 mg/mL. To investigate the inflammation inhibitory effect, we used RAW264.7 cells, these extracts were inhibited inflammatory reaction by suppressing the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in dose-dependent manner. In the assay of HaCaT cells' proliferation, the 24 hr treatment of the extracts significantly accelerarted cell proliferation in the range of concentrations used. The two fractions inhibited the proliferation of Malssezia furfur, the bacteria induce the dandruff. Finally, the CF could significantly inhibit the DHT production at 1, 10, 50, $100{\mu}g/mL$, but EA showed the inhibitory effect at the concentration over $50{\mu}g/mL$. The overall results of this study suggest that the chloroform (CF) and ethylacetate fractions (EA) from P. grandiflorum could be a useful raw material for the hair loss prevention products.

The Chemical Composition of Persimmon (Diospyros kaki, Thumb) Leaf Tea (감 (Diospyros kaki, Thumb) 잎차의 화학 성분)

  • 성낙주;정선영;이수정;조종수;강신권
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.720-726
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    • 1995
  • Chemical components relevant to the characteristic taste of the Korean native persimmon(Diospyros kaki, Thumb) leaf tea were analyzed. Samples were processed by using three different methods ; SHT(steamed and then hot-air dried), DHT(dried in the shade, steamed and then hot-air dried) and RHT(roasted and then hot-air dried). The components analyzed were general compositions of dried perisimmon leaves and extracted solution. The composition of moisture, ash, crude lipid and total nitrogen did not show significant variation among different processing methods of the persimmonleaf tea. The contents of caffeine, tannin and vitamin C in persimmon leaf tea were in the range of $178.4~209.8{\mu}mol/g$, 29.1~38.5mg% and 325.3~2084.7mg%, respectively. The vitamin C content was significantly higher in the RHT than other treatments. The contents of caffeine, tannin and vitamin C in the tea extracted solution were in the range of $101.5~130.1{\mu}mol/g$, 15.4~25.9mg% and 111.0~1274.3mg%, respectively. The vitamine C in the tea solution was the highest in the RHT treatment and 61.1% of vitamin C in the leaf tea was extracted out in these processing methods. The major amino acids contained in the leaf tea were in decreasing order glutamic acid, aspartic acid, leucine and phenylalanine, these four amino acids consisting 38.9~39.8% of the total amino acid contained in the persimmon leaf tea. The major amino acids contained in the tea solution were glutamic acid, proline, histidine and arginine. Six kinds of 5'-nucleotides, CMP, AMP, UMP, IMP, GMP and hypoxanthine were detected and CMP was the most abundant component in fresh leaf, leaf tea and tea solution. The second highest 5'-nucleotides in both leaf tea and tea solutions were GMP, AMP and UMP in all processing method. The highest free sugar contained in the fresh leaf tea and tea solution was sucrose.

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