The Genome-Wide Expression Profile of <i>Nelumbinis semen</i> on Lipopolysaccharide-Stimulated BV-2 Microglial Cells

Sohn, Sung-Hwa,Chung, Hwan-Suck,Ko, Eunjung,Jeong, Hyuk-joon,Kim, Sung-Hoon,Jeong, Jin-Hyun,Kim, Yangseok,Shin, Minkyu,Hong, Moochang,Bae, Hyunsu Pharmaceutical Society of Japan 2009 Biological & pharmaceutical bulletin Vol.32 No.6

<P>This study was conducted to evaluate the protective mechanisms of <I>Nelumbinis semen</I> (NS) on lipopolysaccharide (LPS)-induced activation of BV-2 microglial cells. The anti-inflammatory effects of NS were determined by analyzing nitric oxide production and proinflammatory cytokines using enzyme-linked immunosorbent assay. The mechanism was evaluated in BV-2 cells with or without NS treated with LPS for various lengths of time using oligonucleotide microarray and real time reverse transcription-polymerase chain reaction. The oligonucleotide microarray analysis revealed that mitogen activated protein kinase (MAPK) signaling pathway-related genes such as <I>Fgfr3</I>, <I>Fgf12</I>, <I>Rasal2</I>, <I>Nfkb2</I>, <I>Map2k5</I>, <I>Mapk1</I>, <I>Map3k7</I>, and <I>NFatc2</I> were down-regulated in LPS activated BV-2 cells by pretreatment with NS. In addition, significant decreases in <I>Nos1ap</I> gene expression were observed with NS pretreatment. Cluster linked pathway analysis using the Kyoto Encyclopedia of Genes and Genomes database revealed that the effects of NS were closely associated with the regulation of mitochondria functions. These results suggested that NS can affect the MAPK signaling pathway and mitochondrial functions in BV-2 cells activated with LPS.</P>

Mild stressed rat을 이용한 數種韓藥의 抗憂鬱效果에 대한 실험적 연구

윤정환,신동원,배현수,신민규,홍무창 대한동의생리학회,대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.1

This study was performed to investigate the anti-depression effect of herbal extracts (Rehmanniae Radix Preparat, Corni Fructus, Lycii Fructus, Pinelliae Rhizoma, Nelumbinis Semen) in the chronic mild stressed rat. After adaptation period for 2 weeks, each herbal extract was administered to the rats divided 5 groups. For comparison, St. John's wart was administered to another group in the same environment. The chronic mild stress was induced to rats by overnight illumination for 2 days before the forced swimming test. Struggling time, first latency and rest duration were calculated during the forced swimming test to evaluate the anti-depressant effect. The results were as follows: 1. In the result of evaluation of the struggling time, there was no significance in Pinelliae Rhizoma (-1.1%), Comi Fructus (-3.9%), Rehmanniae Radix Preparat (15.2%), Lycii Fructus (4.9%), St. John's wart (25.2%). There was a significant increase only in Nelumbinis Semen (43.9%) (P-0.0037). 2. In the result of evaluation of the first latency time, there was no significance in Lycii Fructus (-21.37%), Rehmanniae Radix Preparat (38.4%), Pinelliae Rhizoma (65.5%), Corni Fructus (29.2%), St. John's wart (75.8%). There was a significant increase only in Nelumbinis Semen (90.2%) (P=0.0116). 3. In the result of evaluation of the first rest duration, there was no change in St. John's wart, and Decrease was shown in Pinelliae Rhizoma (62.4%), Corni Fructus (31.6%) Rehmanniae Radix Preparat (63.1%), Lycii Fructus (12.4%), Nelumbinis Semen (59.0%), but there was no significance in all. 4. Nelumbinis Semen is considered to have a superior anti-depression effect than the other herbal extracts, and to have a superior anti-depression effect than St. John's wart being used commonly.

Nelumbinis Semen Reverses a Decrease in $5-HT_{1A}$Receptor Binding Induced by Chronic Mild Stress, a Depression-like Symptom

Jang, Choon-Gon,Kang, Moon-Kyu,Cho, Jae-Han,Lee, Sun-Bok,Kim, Hyun-Taek,Park, Soon-Kwon,Lee, Jin-Woo,Park, Seong-Kyu,Hong, Moo-Chang,Shin, Min-Kyu,Shim, In-Sup,Bae , Hyun-Su The Pharmaceutical Society of Korea 2004 Archives of Pharmacal Research Vol.27 No.10

Depression is associated with a dysfunctional serotonin (5-hydroxytryptamine; 5-HT) system. More recently, several lines of evidence suggest that an important factor in the development of depression may be a deficit in the function and expression of $5-HT_{1A}$ receptors. The present study assessed if Nelumbinis Semen (N. s.) had an anti-depression effect through reversing a decrease in $5-HT_{1A}$receptor binding in rats with depression-like symptoms induced by chronic mild stress. Using a $5-HT_{1A}$ receptor binding assay, with a specific $5-HT_{1A}$receptor agonist, 8- OH-DPAT (8-hydroxy-2-(di-n-propylamino) tetralin), the mechanism of the anti-depression effect of N. s. on rats was investigated, and the effects compared with two well-known antidepressants, Hyperium Perforatum (St. Johns Wort) and fluoxetine (Prozac). Animals were divided into five groups: the normal (N) group without chronic mild stress (CMS), the control (C) group under CMS for 8 weeks, the Nelumbinis Semen (N. s.) treatment group under CMS for 8 weeks, the Hyperium Perforatum (H. p.) treatment group under CMS for 8 weeks and finally, the fluoxetine (F) treatment group under CMS for 8 weeks. Each treatment was administered to rats during the last 4 weeks of the 8-week CMS. A sucrose intake test was performed to test the anti-depression effect of N. s. The N. s. treatment significantly reversed the decreased sucrose intake under CMS (P<0.05 compared to control group under CMS). In the CA2 and CA3 regions of the hippocampus, both N. s. and H. p. reversed the CMS-induced decrease in $5-HT_{1A}$receptor binding. In the I to II regions of the frontal cortex, N. s. and H. p. also reversed the CMS-induced decrease in$5-HT_{1A}$receptor binding, and even showed a significant increase in $5-HT_{1A}$receptor binding compared to the F treatment group (N. s. vs. P, p<0.05, H. p. vs. P, p<0.05). However, in the hypothalamus, all treatments reversed the CMSinduced decrease in $5-HT_{1A}$receptor binding. This reversal effect of N. s. on the decrease in $5-HT_{1A}$receptor binding in the frontal cortex, hippocampus and hypothalamus of rat brains was very similar to that of H. p, but different from that of F. It is concluded that N. s. presents an anti-depression effect through enhancing $5-HT_{1A}$receptor binding.

Effects of Nelumbinis Semen on Contractile Dysfunction in Ischemic and Reperfused Rat Heart

Jong-Hoon Kim,강문규,Chongwoon Cho,정환석,Chang-Woon Kang,Shoukat Parvez,배현수 대한약학회 2006 Archives of Pharmacal Research Vol.29 No.9

Nelumbinis Semen (NS), or lotus seed, is one of the most well-known traditional herbal medicines and is frequently used to treat cardiovascular symptoms in Korea. The anti-ischemic effects of NS on ischemia-induced isolated rat heart were investigated through analyses of changes in blood pressure, aortic flow, coronary flow, and cardiac output. The subjects in this study were divided into two groups: a control, untreated ischemia-induced group, and an ischemia-induced group treated with NS. There were no significant differences in perfusion pressure, aortic flow, coronary flow and cardiac output between the groups before ischemia was induced. The supply of oxygen and buffer was stopped for ten minutes to induce ischemia in isolated rat hearts, and NS was administered during ischemia induction. NS treatment significantly prevented decreases in perfusion pressure, aortic flow, coronary flow and cardiac output under ischemic conditions (p<0.01). In addition, the mechanism of the anti-ischemic effects of NS was also examined through quantitation of intracellular calcium content in rat neonatal cardiomyocytes. NS significantly prevented intracellular calcium increases induced by isoproterenol (p<0.01). These results suggest that NS has distinct anti-ischemic effects through calcium antagonism.

Nelumbinis Semen Reverses a Decrease in 5-HT_(1A) Receptor Binding Induced by Chronic Mild Stress, a Depression-like Symptom

Jang, Choon-Gon,Kang, Moonkyu,Cho, Jae-Han,Lee, Sun-Bok,Kim, Hyuntaek,Park, Soonkwon,Lee, Jinwoo,Park, Seong-Kyu,Hong, Moochang,Shin, Min Kyu,Shim, In-Sup,Bae, Hyunsu WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2004 東西醫學硏究所 論文集 Vol.2004 No.-

Depression is associated with a dysfunctional serotonin (5-hydroxytryptamine; 5-HT) system. More recently, several lines of evidence suggest that an important factor in the development of depression may be a deficit in the function and expression of 5-HT_(1A) receptors. The present study assessed if Nelumbinis Semen (N. s.) had an anti-depression effect through reversing a decrease in 5-HT_(1A) receptor binding in rats with depression-like symptoms induced by chronic mild stress. Using a 5-HT_(1A) receptor binding assay, with a specific 5-HT_(1A) receptor agonist, 8-OH-DPAT (8-hydroxy-2-(di-n-propylamino) tetralin), the mechanism of the anti-depression effect of N. s. on rats was investigated, and the effects compared with two well-known antidepressants, Hyperium Perforatum (St. Johns Wort) and fluoxetine (Prozac). Animals were divided into live groups: the normal (N) group without chronic mild stress (CMS), the control (C) group under CMS for 8 weeks, the Nelumbinis Semen (N. s.) treatment group under CMS for 8 weeks, the Hyperium Perforatum (H. p.) treatment group under CMS for 8 weeks and finally, the fluoxetine (F) treatment group under CMS for 8 weeks. Each treatment was administered to rats during the last 4 weeks of the 8-week CMS. A sucrose intake test was performed to test the anti-depression effect of N. s. The N. s. treatment significantly reversed the decreased sucrose intake under CMS (P<0.05 compared to control group under CMS). In the CA2 and CA3 regions of the hippocampus, both N. s. and H. p. reversed the CMS-induced decrease in 5-HT_(1A) receptor binding. In the Ⅰ to II regions of the frontal cortex, N. s. and H. p. also reversed the CMS-induced decrease in 5-HT_(1A) receptor binding, and even showed a significant increase in 5-HT_(1A) receptor binding compared to the F treatment group (N. s. vs. P₁ p<0.05, H. p. vs. P₁ p<0.05). However, in the hypothalamus, all treatments reversed the CMS-induced decrease in 5-HT_(1A) receptor binding. This reversal effect of N. s. on the decrease in 5-HT_(1A) receptor binding in the frontal cortex, hippocampus and hypothalamus of rat brains was very similar to that of H. p, but different from that of F. It is concluded that N. s. presents an anti-depression effect through enhancing 5-HT_(1A) receptor binding.

Nelumbinis Semen Reverses a Decrease in 5-HT1A Receptor Binding Induced by Chronic Mild Stress, a Depression-like Symptom

장춘곤,강문규,Jae-Han Cho,Sun-Bok Lee,Hyuntaek Kim,박순권,Jinwoo Lee,박성규,Moochang Hong,Min Kyu Shin,심인섭,배현수 대한약학회 2004 Archives of Pharmacal Research Vol.27 No.10

Depression is associated with a dysfunctional serotonin (5-hydroxytryptamine; 5-HT) system. More recently, several lines of evidence suggest that an important factor in the development of depression may be a deficit in the function and expression of $5-HT_{1A}$ receptors. The present study assessed if Nelumbinis Semen (N. s.) had an anti-depression effect through reversing a decrease in $5-HT_{1A}$receptor binding in rats with depression-like symptoms induced by chronic mild stress. Using a $5-HT_{1A}$ receptor binding assay, with a specific $5-HT_{1A}$receptor agonist, 8- OH-DPAT (8-hydroxy-2-(di-n-propylamino) tetralin), the mechanism of the anti-depression effect of N. s. on rats was investigated, and the effects compared with two well-known antidepressants, Hyperium Perforatum (St. Johns Wort) and fluoxetine (Prozac). Animals were divided into five groups: the normal (N) group without chronic mild stress (CMS), the control (C) group under CMS for 8 weeks, the Nelumbinis Semen (N. s.) treatment group under CMS for 8 weeks, the Hyperium Perforatum (H. p.) treatment group under CMS for 8 weeks and finally, the fluoxetine (F) treatment group under CMS for 8 weeks. Each treatment was administered to rats during the last 4 weeks of the 8-week CMS. A sucrose intake test was performed to test the anti-depression effect of N. s. The N. s. treatment significantly reversed the decreased sucrose intake under CMS (P<0.05 compared to control group under CMS). In the CA2 and CA3 regions of the hippocampus, both N. s. and H. p. reversed the CMS-induced decrease in $5-HT_{1A}$receptor binding. In the I to II regions of the frontal cortex, N. s. and H. p. also reversed the CMS-induced decrease in$5-HT_{1A}$receptor binding, and even showed a significant increase in $5-HT_{1A}$receptor binding compared to the F treatment group (N. s. vs. P, p<0.05, H. p. vs. P, p<0.05). However, in the hypothalamus, all treatments reversed the CMSinduced decrease in $5-HT_{1A}$receptor binding. This reversal effect of N. s. on the decrease in $5-HT_{1A}$receptor binding in the frontal cortex, hippocampus and hypothalamus of rat brains was very similar to that of H. p, but different from that of F. It is concluded that N. s. presents an anti-depression effect through enhancing $5-HT_{1A}$receptor binding.

LPS로 자극한 BV-2 microglial cell에서 Microarray를 통한 蓮子肉의 유전자 발현 분석

김수오(Kim Soooh),임병묵(Lim Byungmook),배현수(Bae Hyunsu) 한국한의학연구원 2008 한국한의학연구원논문집 Vol.14 No.1

Nelumbinis Semen(NS) has been used in traditional medicine to treat diseases such as depression and diarrhea. In inflammatory responses, microglia produces molecules which are known to play roles in the central nervous system. And we previously studied NS inhibited nitric oxide synthase and secretion of tumor necrosis factor alpha. To explore the global gene expression profiles in BV-2 microglial cell line treated with NS, microarray analysis was performed. The cells were treated with LPS or NS plus LPS for 30min, 1h, 3h, and 6h, respectively. Of 45,101 known genes, with cutoff value of 3-fold change in the expression, 340, 644, 280 and 219 genes were upregulated and 503, 570, 694 and 484 were downregulated in NS treated cells at each time point. The results of the present study shows that treatment of NS reversed the LPS-induced upregulation of such genes as Ecoxsackievirus and adenovirus receptor(CAR), pellino 1, and S100P binding protein. It is thought that microarrays will play an ever-growing role in the advance of our understanding of the pharmacologic actions NS.

연자육의 페놀성 성분 및 Tyrosinase 저해 활성

정지연,모은진,황방연,이미경 한국생약학회 2015 생약학회지 Vol.46 No.1

In the course of screening tyrosinase inhibitory activity, EtOAc-soluble fraction of Nelumbinis Semen (Seeds of Nelumbo nucifera) showed significant inhibition. Further fractionation of the EtOAc-soluble fraction resulted in 12 compounds, which were identified as 4-(hydroxymethyl)phenol (1), tyrosol (2), 4-(hydroxymethyl)benzaldehyde (3), 4-hydroxybenzoic acid (4), 4-(2-methoxyvinyl)benzene-1,2-diol (5), 2,6-dihydroxybenzoic acid (6), (2R-trans)-2,3-dihydro-3,5,7,8-tetrahydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one (7), (+)-catechin (8), elephantorrhizol (9), (+)-dehydrovomifoliol (10), (-)-boscialin (11) and uridine (12). Compounds 5 and 7 were first reported from this plant. Among the isolated compounds, compound 7 showed strong inhibition on tyrosinase activity with mixed mechanism of competitive and noncompetitive inhibition.

연자육의 항우울 효과에 대한 뇌부위별 신경전달물질의 변화

김지현(Ji Hyun Kim),이규섭(Kyu-Sop Lee),정혜영(Hye-Young Joung  ),심현수(Hyun Soo Shim),배현수(Hyunsu Bae),김경수(Kyung Soo Kim),심인섭(Insop Shim) 대한스트레스학회 2008 스트레스硏究 Vol.16 No.4

선행 연구에서 연자육 추출물 중 강제수영 검사를 통해 항우울 효과를 검증하여 가장 뛰어난 결과를 보인 NS37 (n-BuOH 추출물의 추가 분획물, H2O:MeOH=3:7)의 항우울 효과 기전을 규명하고자, HPLC-ECD 시스템을 이용하여 뇌의 우울증 신경회로로 알려진 대뇌피질-시상하부-해마의 부위별 세로토닌과 도파민의 함량을 조사하고 이들 대사산물의 측정 및 대사속도를 검토하였다. 연구 결과 NS37-10 (10 mg/kg)군에서 대뇌피질, 시상하부, 해마에서 도파민의 유의한 증가 및 시상하부, 해마에서 도파민 교체율의 유의한 감소를 나타냄으로써 항우울 효과를 확인하였다. In the previous study, we examined that a variety of extracts of Nelumbinis semen (NS) have an antidepressant effect on the forced swimming test (FST) in the rat, which is widely used as a behavioral screening test for antidepressant activity of new compounds. Depression-like symptom in n-BuOH extract showed the strongest inhibitory effect on the immobility time and further fractionated n-BuOH with H2O and MeOH (3:7) fraction (NS37) was the most effective on the immobility time on FST. The purpose of the present study was to observe the neurochemical changes on the anti-depressant actions of unknown compounds in the NS37 extract by using HPLC coupled with electrochemical detection (ECD). The results showed that the NS37-10 (10mg/kg) has a significant increase of dopamine in cortex, hypothalamus, and hippocampus and a significant decrease of dopamine turnover in hypothalamus and hippocampus. These neurochemical actions of NS37 extract could be responsible for the beneficial effect on depression. (Korean J Str Res 2008;16:393∼400)

연자육의 6-하이드록시도파민으로 유도된 도파민 세포 독성에 대한 보호효과

김효근 ( Hyo Geun Kim ),오명숙 ( Myung Sook Oh ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.2

Objectives: This study was performed to evaluate the neuroprotective effect of water extracts from Nelumbinis semen (NSW) in dopaminergic cells. Methods: We performed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azinobis- 3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of NSW. We also evaluated the neuroprotective effects against 6-hydroxydopamine (6-OHDA)-induced toxicity using 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide assay (MTT) assay, trypan blue cytotoxicity assay, and nitric oxide assay in SH-SY5Y cells and tyrosine hydroxylase (TH) immunohistochemistry in primary rat dopaminergic neurons. Results: NSW showed IC50 values of 184.80 and 92.90 μg/mL in DPPH and in ABTS assays, respectively. NSW showed 1.05% of total polyphenol contents. NSW showed protective effect against 6-OHDA-induced neurotoxicity whereas no influence on cell viability at the concentration of 1~50 μg/mL. NSW reduced NO generation while 6-OHDA produced it. Moreover, it protected rat dopaminergic neurons against 6-OHDA at a dose of 1 μg/mL. Conclusions: These results indicated that NSW has neuroprotective effect against 6-OHDA-induced neurotoxicity through antioxidant activity in dopaminergic cell culture.