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SPECT Imaging of Dopamine Transporter with [I-123] IPT in Normal Controls and Parkinson's Patients (정상인과 파킨슨병 환자에서 [I-123]IPT SPECT를 이용한 도파민 재섭취부위의 영상화)

  • Sohn, Hyung Sun;Kim, Euy Neyng;Lee, Kyung Jin;Rha, Hyung Keun;Son, Byung Chul;Choi, Chang Rhack
    • Journal of Korean Neurosurgical Society
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    • v.30 no.3
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    • pp.342-348
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    • 2001
  • Objective : Dopamine transporter concentrations have been known to decrease in Parkinson's disease(PD). The aim of the present study was to evaluate the correlation between SPECT measurements of [I-123]N-(3-iodopropene-2-yl)-$2{\beta}$-carbomethoxy-$3{\beta}$-(4-chlorophenyl) tropane(IPT) as an imaging agent for measuring changes in transporter concentrations with PD. Patients and Methods : IPT labelled with $4.87{\pm}1.29mCi$($180.19{\pm}47.73MBq$) of [I-123] was intravenously injected into 23 patients(age : $58{\pm}12$) with PD and three normal controls(NC)(age : $37{\pm}7$) as bolus. Brain SPECT were then performed at 1 hour and 2 hours after injection on a double headed camera. The statistical parameters were the contrast ratio of left basal ganglia(BG) and right basal ganglia to occipital cortex(OCC) per milli curies of injected radiotracer at 1 hour and 2 hours. The correlations were evaluated between these parameters and Hoehn-Yahr classification of the patients. Results : The(BG - OCC)/OCC/mCi ratios at 1 hour and 2 hours for PD and NC were $0.14{\pm}0.07$ and $0.27{\pm}0.07$(1 hour) and $0.12{\pm}0.07$ and $0.34{\pm}0.04$(2 hour), respectively. The(BG - OCC)/OCC/mCi ratios of Parkinson's disease were decreased with higher grade of Hoehn-Yahr classification of the patients. The ratio between BG and OCC for PD were clearly separated from NC and may be useful outcome measures for clinical diagnosis. Conclusion : The findings suggest that IPT may be a very useful tracer for early diagnosis and treatment of PD and study of dopamine re-uptake site.

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Inhibitory Effects of the Seed Extract of Myristicae Semen on the Proliferation of Human Tumor Cell Lines (II) (육두구 추출물의 암세포증식 저해 효과 (제 2보))

  • Lee, Jung-Won;Choi, Yeon-Hee;Yoo, Mi-Young;Choi, Sang-Un;Hong, Kyung-Sik;Lee, Byung-Hoi;Yon, Gyu-Hwan;Kim, Young-Sup;Kim, Young-Kyoon;Ryu, Shi-Yong
    • Korean Journal of Pharmacognosy
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    • v.37 no.3
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    • pp.206-211
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    • 2006
  • The methanol extract from seed of Myristica fragrans (Myristicaceae) demonstrated a potent inhibition on the pro-liferation of cultured human tumor cells such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCT-15 (colon) in vitro. By the continuous effort to purify the active components responsible far the anti-proliferative effect on tumor cell lines, we have isolated eleven kinds of lignan components, i. e., safrole (1), machilin A (2), licarin B (3), macelignan (4), mere-dihydroguaiaretic acid (5), mγnstargenol A (6), methoxyeugenol (7), machilin F (8), licarin A (9), nectandrin B (10), and 2-(4-allyl-2,6-dimethoxyphenoxy)-1-(4-hydroxy-3-methoxyphenyl)propan-1-ol (11) together with a novel furan fatty acid, (E)-3-(3-methyl-5-pentylfuran-2-yl) acrylic acid (12) from seed extract of M. fragrans. Chemical structures of the isolated components (1-12) were established bγ the aid of NMR spectroscopic analyses, i. e., COSY HMQC and HMBC. Each of the Isolates demonstrated a potent inhibition on the proliferation of cultured human tumor cells such as A549 (non small cell lung), SK-OY-3 (ovary), SK-MEL-2 (melanoma) and HCT-15 (colon) in vitro.

Treatment with Rutin - A Therapeutic Strategy for Neutrophil-Mediated Inflammatory and Autoimmune Diseases - Anti-inflammatory Effects of Rutin on Neutrophils -

  • Nikfarjam, Bahareh Abd;Adineh, Mohtaram;Hajiali, Farid;Nassiri-Asl, Marjan
    • Journal of Pharmacopuncture
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    • v.20 no.1
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    • pp.52-56
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    • 2017
  • Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor $(TNF)-{\alpha}$ productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin ($25{\mu}M$) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the $TNF-{\alpha}$, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin ($1-100{\mu}M$), after which MTT was appended and incubated at $37^{\circ}C$ for 4 hour. Results: Rutin at concentrations up to $100{\mu}M$ did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and $TNF-{\alpha}$ productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and $TNF-{\alpha}$ productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/autoimmune diseases.

Overexpression of Rcan1-1L Inhibits Hypoxia-Induced Cell Apoptosis through Induction of Mitophagy

  • Sun, Lijun;Hao, Yuewen;An, Rui;Li, Haixun;Xi, Cong;Shen, Guohong
    • Molecules and Cells
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    • v.37 no.11
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    • pp.785-794
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    • 2014
  • Mitophagy, a cellular process that selectively targets dysfunctional mitochondria for degradation, is currently a hot topic in research into the pathogenesis and treatment of many human diseases. Considering that hypoxia causes mitochondrial dysfunction, which results in cell death, we speculated that selective activation of mitophagy might promote cell survival under hypoxic conditions. In the present study, we introduced the Regulator of calcineurin 1-1L (Rcan1-1L) to initiate the mitophagy pathway and aimed to evaluate the effect of Rcan1-1L-induced mitophagy on cell survival under hypoxic conditions. Recombinant adenovirus vectors carrying Rcan1-1L were transfected into human umbilical vein endothelial cells and human adult cardiac myocytes. Using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay and Trypan blue exclusion assay, Rcan1-1L overexpression was found to markedly reverse cell growth inhibition induced by hypoxia. Additionally, Rcan1-1L overexpression inhibited cell apoptosis under hypoxic conditions, as detected by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay. Meanwhile, the mitochondria-mediated cell apoptotic pathway was inhibited by Rcan1-1L. In contrast, knockdown of Rcan1-1L accelerated hypoxia-induced cell apoptosis. Moreover, Rcan1-1L overexpression significantly reduced mitochondrial mass, decreased depolarized mitochondria, and downregulated ATP and reactive oxygen species production. We further delineated that the loss of mitochondrial mass was due to the activation of mitophagy induced by Rcan1-1L. Rcan1-1L overexpression activated autophagy flux and promoted translocation of the specific mitophagy receptor Parkin into mitochondria from the cytosol, whereas inhibition of autophagy flux resulted in the accumulation of Parkin-loaded mitochondria. Finally, we demonstrated that mitochondrial 1permeability transition pore opening was significantly increased by Rcan1-1L overexpression, which suggested that Rcan1-1L might evoke mitophagy through regulating mitochondrial permeability transition pores. Taken together, we provide evidence that Rcan1-1L overexpression induces mitophagy, which in turn contributes to cell survival under hypoxic conditions, revealing for the first time that Rcan1-1L-induced mitophagy may be used for cardioprotection.

Whitening Activity of Sambucus Sieboldiana Var. Pendula (Nakai) Extract (말오줌나무 추출물의 미백활성 검증)

  • Yoo, Dan-Hee;Kim, Jin-Tae;Oh, Min-Jeong;Yeom, Hyeon-Ji;Lee, Jin-Young
    • Journal of Life Science
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    • v.29 no.3
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    • pp.279-286
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    • 2019
  • This study evaluated the anti-oxidant and whitening effects of a 70% ethanol extract of the Sambucus sieboldiana var. pendula (Nakai) (SS). At $1,000{\mu}g/ml$ concentration, the electron donating ability of this SS extract was found to be 86.21% and the ABTS+ radical scavenging ability was 97.9%. In terms of whitening activity, the tyrosinase inhibitory effect of the extract was 37%, also at $1,000{\mu}g/ml$ concentration. To explore the extractefftoxicity to B16F10 melanoma cells, a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide assay was performed. Results showed 90% or more cells remained viable at $100{\mu}g/ml$ concentration. A Western blot of the SS extract was used to measure microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2), and the tyrosinase protein expression inhibitory effect at 25, 50, $100{\mu}g/ml$ concentrations; ${\beta}-actin$ was used as a positive control. Consequently, the MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect were seen to decrease by 34.5%, 45.6%, 58.4%, and 79.6%, respectively, at $100{\mu}g/ml$ concentration. These were also then measured by reverse transcription-polymerase chain reaction at 25, 50, $100{\mu}g/ml$ concentrations with GAPDH as a positive control. As a result, the SS extract was seen to decrease MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect by 85.4%, 67.5%, 85.2%, 67.1%, respectively at the $100{\mu}g/ml$ concentration. We therefore confirmed the possibility of Sambucus sieboldiana var. pendula (Nakai) extract as a whitening material.

Anti-inflammatory effect potentials of ethanol extracts from fermentated Caryopteris incana by Lactobacillus plantarum on induced to LPS with Raw 264.7 cell (LPS로 유도된 Raw 264.7 cell에서 Lactobacillus plantarum 발효가 층꽃나무(Caryopteris incana) 에탄올 추출물의 염증반응에 미치는 영향)

  • Park, Mi-Jeong;Park, Hye-Jin;Lee, Eun-Ho;Jung, Hee-Young;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.61 no.2
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    • pp.141-150
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    • 2018
  • In this study, the inflammation of ethanol extracts from Caryopteris incana (CI) and fermented C. incana (FCI) on induced to lipopolysaccharide with Raw 264.7 cell was tested. The composition profile of L. plantarum was changed by fermentation, and confirmed by HPLC analysis. We performed the 3-[4,5-dimethylthiazol]-2-yl]-2,5-diphenyltetrazolium bromide assay to evaluate the toxicity of CI and FCI extracts. In cell viability, cell toxicity was not shown at 5, 10 and $15{\mu}g/mL$ of CI extracts and 10, 20, 30 and $40{\mu}g/mL$ of FCI extracts. The results of inducible nitric oxide synthase and cyclooxygenase-2 protein production were confirmed to be inhibitory in a concentration-dependent manner, respectively. Additionally, protein expression of nitric oxide and prostaglandin $E_2$ by CI and FCI extracts were also inhibited in a concentration-dependent manner. In the result of pro-inflammatory cytokine, $15{\mu}g/mL$ concentration of CI extracts was showed tumar necrosis factor $(TNF)-{\alpha}$ (57.3%), interleukin (IL)-6 (35.2%), and $IL-1{\beta}$ (48.0%), respectively. And $40{\mu}g/mL$ of FCI extracts was showed $TNF-{\alpha}$ (34.6%), IL-6 (32.1%), and $IL-1{\beta}$ (30.0%), respectively. These results suggest that FCI extracts showed better effect of anti-inflammatory than CI extracts. Therefore, it was found that both CI and FCI can be used as an excellent material for the development of new anti-inflammatory resource.

Dopamine Transporter Density of the Basal Ganglia in Children with Attention Deficit Hyperactivity Disorder Assessed with I-123 IPT PECT (I-123 IPT SPECT를 이용한 주의력결핍 과잉행동장애 아동에서의 methylphenidate 투여 전후의 기저 신경절 도파민 운반체 밀도 변화 측정)

  • Ryu, Won-Gee;Kim, Tae-Hoon;Ryu, Young-Hoon;Yun, Mi-Jin;Cheon, Keun-Ah;Chi, Dae-Yoon;Kim, Jong-Ho;Choi, Tae-Hyun;Lee, Jong-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.4
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    • pp.235-244
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    • 2003
  • Purpose: Attention deficit hyperactivity disorder (ADHD) has been known as psychiatric disorder in childhood associated with dopamine dysregulation. In present study, we investigated changes in dopamine transporter (DAT) density of the basal ganglias using I-123 N-(3-iodopropen-2-yl) -2-carbomethoxy-3beta-(4-chlorophenyl) tropane [I-123 IPT] SPECT in children with ADHD before and after methylphenidate treatment. Materials and Method: Nine drug-naive children with ADHD and seven normal children were included in the study. We peformed brain SPECT two hours after the intravenous administration of I-123 IPT and made both quantitative and qualitative analyses using the obtained SPECT data, which were reconstructed for the assessment of soecific/nonspecific DAT binding ratios in the basal ganglia. All children with ADHD reperformed [123I]IPT SPECT after treatment with methylphenidate(0.7mg/kg/d) during about 8 weeks. SPECT data reconstructed for the assessment of specific/nonspecific DAT binding ratio of the basal ganglia were compared between before and after treatment methylphenidate. We investigated correlation between the change of ADHD symptom severity assessed with ADHD rating scale-IV and specific/nonspecific DAT binding ratio of basal ganglia. Results: Children with ADHD had a significantly greater specificinonspecific DAT binding ratio of the basal ganglia comparing to normal children(Right : z = 2.057, p = 0.041 : Left : z : 2.096, p = 0.032). Under treatment with methylphenidate in all children with ADHD, specificinonspecific DAT binding ratio of both basal ganglia decreased significantly greater than before treatment with methylphenidate (Right : t = 3.239, p = 0.018 ; Left : t = 3.133, p = 0.020). However, no significant correlation between the change of ADHD symptom severity scores and specific/nonspecific DAT binding ratio of the basal ganglia were found. Conclusions: These findings support the complex dysregulation of the dopaminergic neurotransmitter system in children with ADHD.

Preventive Characteristics of Garlic Extracts Using in vitro Model System on Alzheimer's Disease (In vitro model system을 활용한 마늘 추출물의 치매예방 특성)

  • Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Jeong, Chang-Ho;Jeong, Hee-Rok;Shin, Jung-Hye;Kang, Min-Jung;Sung, Nak-Ju;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.4
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    • pp.45-55
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    • 2010
  • In this study, the acetylcholinesterase (AChE) inhibition and neuronal cell protective effects of water, 100% methanol and dichlromethane extracts from garlic were investigated. We found that dichloromethane extract of garlic resulted in a dose-dependent manner on AChE inhibition ($IC_{50}$: $36.1{\mu}g/mL$). In cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), cell viabilities of water, 100% methanol and dichlromethane extracts were lower (almost under 40%) than amyloid ${\beta}$ protein ($A{\beta}$)-induced neurotoxicity. Because $A{\beta}$ is also known to increase neuronal cell membrane breakdown, neuronal apoptosis was further confirmed by lactate dehydrogenase (LDH) and neutral red uptake (NRU) assay. Water extract presented relative protection against $A{\beta}$-induced membrane damage in LDH assay. However all garlic extracts showed significant problem with decrease of cell viability in NRU assay, especially at dichloromethan extract. To determine active compounds in column fractions (98:2 fraction) from dichloromethane extract which showed significant AChE inhibitory effect, we performed HPLC and LC-MS analysis. It was supposed that garlic may contain allyl methyl disulfide, diallyl monosulfide, and diallyl disulfide as active compounds.

The Effect of Woogakseungmatang Extract on NO Production in LPS- Stimulated RAW 264.7 Cells (우각승마탕이 LPS로 유도된 RAW 264.7 세포에서 NO 생산에 미치는 영향)

  • Jo, Na Young
    • Korean Journal of Acupuncture
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    • v.35 no.4
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    • pp.166-173
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    • 2018
  • Objectives : Woogakseungmatang is a prescription medication mainly used to treat facial paralysis in Korean medicine. The purpose of this study is to investigate the effects of Woogakseungmatang on anti-inflammation and anti-oxidation. Methods : Woogakseungmatang was extracted using hot water. Cytotoxicity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) method; nitric oxide(NO) production and Prostaglandin $E_2$ ($PGE_2$) production in RAW cells treated with Woogakseungmatang were investigated; and the cytokine changes associated with inflammation were examined. The antioxidant capacity of Woogakseungmatang was measured using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Results : RAW cells treated with Woogakseungmatang showed 90% cell viability at a $100-{\mu}g/ml$ concentration. NO production was decreased by 15% at a $100-{\mu}g/ml$ concentration. $PGE_2$ production was decreased by 18% at a $100-{\mu}g/ml$ concentration. Interleukin $1{\beta}$ ($IL-1{\beta}$), interleukin 6(IL-6), and tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) were significantly reduced at $100{\mu}g/ml$ compared with those in the control group. The DPPH free radical scavenging capability was more than 50% at $100{\mu}g/ml$. Conclusions : Woogakseungmatang showed only a slight anti - inflammatory effect at $100{\mu}g/ml$ and it was difficult to confirm the concentration-dependent anti-inflammatory effect. Therefore, this study means to confirm the potential anti-inflammatory effects of Woogakseungmatang. Based on this research, more systematic and diverse studies should be conducted.