멜라토닌이 랫트에서 심박수에 미치는 영향

심소연,신세린,김진상,Shim, So-yeon,Shin, Se-rin,Kim, Jin-shang 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.4

Evidence from the last 10 years have been suggested that melatonin mainly produce a depressant effect on the cardiac system, but we found an activating effect of melatonin on heart rate in this research. To determine the hypothesis that melatonin has dual effects on physiological behaviour of cardiac system, we investigated the effects of melatonin on heart rate in isolated rat atria and anesthetized rats. Regardless of concentration, melatonin produced bradycardia in the 84 cases of 148 experiments (57 %) and tachycardia in the 64 cases of 148 experiments (43 %). And in atrium, melatonin produced a decrease automaticity in 52 cases of 86 experiments (60 %) and increase automaticity in 40 % (34/86 cases). Also, these effects are not significnat relationship with concetration of melatonin. The melatonin-induced bradycardia in vivo was inhibited by pretreatment of atropine or bilateral cervical vagotomy. Also, in isolated atrium the melatonin-induced decrease in automaticity was inhibited by pretreatment of atropine. These melatonin-induced responses were potenitated by pretreatment of propranolol. The melatonin-induced tachycardia in vivo was inhibited by pretreatment of propranolol, nifedipine or bilateral cervical vagotomy, but not by pretreatment of atropine. The melatonin-induced incease in automaticity in isolated atrium was converted to decrease in automaticity by pretreatment of propranolol. In addition, the change in heart rate caused by adrenoceptor agonists was inhibited by pretreatment of melatonin. These results indicate that melatonin-induced bradycardia may be related to a muscarinic receptor activation and melatonin-induced tachycardia may be related to a $\beta$-adrenoceptor stimulation.

Suppression of Osteoclastogenesis by Melatonin: A Melatonin Receptor-Independent Action

Kim, Hyung Joon,Kim, Ha Jin,Bae, Moon-Kyoung,Kim, Yong-Deok MDPI 2017 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.18 No.6

<P>In vertebrates, melatonin is primarily secreted from the pineal gland but it affects various biological processes including the sleep-wake cycle, vasomotor control, immune system and bone homeostasis. Melatonin has been known to promote osteoblast differentiation and bone maturation, but a direct role of melatonin on osteoclast differentiation is still elusive. The present study investigated the effect of melatonin on the differentiation of macrophages to osteoclasts. The presence of melatonin significantly reduced receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis and the siRNA-mediated knockdown of the melatonin receptor failed to overcome the anti-osteoclastogenic effect of melatonin. Although melatonin treatment did not affect the phosphorylation of extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK), it markedly inhibited the activation of NF-κB and subsequent induction of nuclear factor of activated T cell cytoplasmic 1(NFATc1). Thus, our results suggest that melatonin could suppress osteoclast differentiation through downregulation of NF-κB pathway with concomitant decrease in the NFATc1 transcription factor induction. Furthermore, melatonin seems to have an anti-osteoclastogenic effect independent of plasma membrane melatonin receptors. In addition to previously reported properties of melatonin, our study proposes another aspect of melatonin and bone homeostasis.</P>

Antioxidant and Anti-Apoptotic Effect of Melatonin on the Vestibular Hair Cells of Rat Utricles

김정범,정재윤,안진철,이정구,황희준 대한이비인후과학회 2009 Clinical and Experimental Otorhinolaryngology Vol.2 No.1

Objectives. Aminoglycosides are commonly used antibiotic agents, and they are known to generate free oxygen radicals within the inner ear and to cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Melatonin, a pineal secretory product, has the properties of being a powerful direct and indirect antioxidant. The aim of the present study was to prove the antioxidant effect of melatonin against gentamicin-induced ototoxicty. Methods. The utricular maculae of Sprague-Dawley rats were prepared from postnatal day 2-4, and these maculae were were divided into 6 groups as follows: 1) control, 2) melatonin only, 3) gentamicin only, and 4), 5), and 6) gentamicin plus melatonin (10, 50, and 100 μM, respectively). To count the number of hair cells, 5 utricles from each group were stained with phalloidin-FITC on the 1st, 4th, and 7th days after drug administration. Reactive oxygen species (ROS) was assessed by using the fluorescent probe hydrofluorescent diacetate acetyl ester. The caspase-3 activity was also examined with using the fluorescent caspase-3 substrate and performing Western blotting. Results. The result of this study showed that gentamicin induced the loss of utricular hair cells, and this loss of hair cells was significantly attenuated by co-administration of melatonin. Melatonin reduced ROS production and caspase-3 activation in the gentamicin treated utricular hair cells. Conclusion. Our findings conclusively reveal that melatonin has protective effects against gentamicin-induced hair cell loss in the utricles of rat by inhibiting both ROS production and caspase-3 activity. Objectives. Aminoglycosides are commonly used antibiotic agents, and they are known to generate free oxygen radicals within the inner ear and to cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Melatonin, a pineal secretory product, has the properties of being a powerful direct and indirect antioxidant. The aim of the present study was to prove the antioxidant effect of melatonin against gentamicin-induced ototoxicty. Methods. The utricular maculae of Sprague-Dawley rats were prepared from postnatal day 2-4, and these maculae were were divided into 6 groups as follows: 1) control, 2) melatonin only, 3) gentamicin only, and 4), 5), and 6) gentamicin plus melatonin (10, 50, and 100 μM, respectively). To count the number of hair cells, 5 utricles from each group were stained with phalloidin-FITC on the 1st, 4th, and 7th days after drug administration. Reactive oxygen species (ROS) was assessed by using the fluorescent probe hydrofluorescent diacetate acetyl ester. The caspase-3 activity was also examined with using the fluorescent caspase-3 substrate and performing Western blotting. Results. The result of this study showed that gentamicin induced the loss of utricular hair cells, and this loss of hair cells was significantly attenuated by co-administration of melatonin. Melatonin reduced ROS production and caspase-3 activation in the gentamicin treated utricular hair cells. Conclusion. Our findings conclusively reveal that melatonin has protective effects against gentamicin-induced hair cell loss in the utricles of rat by inhibiting both ROS production and caspase-3 activity.

신생쥐에서 유발시킨 저산소 허혈 뇌손상에 대한 melatonin과 저체온치료 병용요법의 효과

박재현,김천수,이상락,이성룡 대한신생아학회 2014 Neonatal medicine Vol.21 No.2

Purpose: Melatonin is a naturally occurring hormone produced by the pineal gland. Melatonin has many pharmacological effects in different tissues or organs. Melatoninis especially known to have antioxidant and neuroprotective effects. Hypothermia is atherapeutic tool against hypoxia-ischemia (HI) of the brain. This study examines theeffect of combined therapy using melatonin and hypothermia in neonatal rats with HI. Methods: Seven-day old rats were subjected to HI and randomized into four groups :vehicle, melatonin alone, vehicle and hypothermia, and melatonin and hypothermia. Melatonin (30 mg/kg) was intraperitoneally administered in two doses: immediatelyfollowing HI, and 24 h later. Hypothermia consisted of whole-body cooling (3 hours,27℃). Sham-treated animals not subjected to HI were also studied. P10, P14, andP35 rats were sacrificed for experiments. Results: Vehicle-treated P10 rats increased in brain infarction compared to controlsin TTC staining study. And also, P35 rats decreased in brain volume of injured hemispherein H&E stain. Melatonin or hypothermia alone did not show any protectiveeffect against HI. However, a combination of melatonin and hypothermia effectivelyreduced the brain injury. In addition, the results of in situ zymography, TUNEL assayand immunofluorescence studies showed that neuroprotective effects were achievedonly with combined therapy. Conclusion: Melatonin may contribute to synergistic effects to neuroprotection ofhypothermia on brain damage after HI. 목적: Melatonin은 송과샘에서 분비되는 호르몬으로, 여러 조직에 있어 다양한 약리작용을 나타낸다. 특히, melatonin은 우수한 항산화 효과와 신경 보호작용을 가진다고 알려져 있다. 저체온치료는 저산소 허혈 뇌손상에 적용되는임상 치료법이다. 이 연구는 저산소 허혈을 유발시킨 신생쥐에서 melatonin 및 저체온치료의 병합요법의 효과에대하여 알아보고자 하였다. 방법: 생후 7일된 쥐에 저산소 허혈을 유발하였고, 실험군은 vehicle군과 melatonin 단독투여군, vehicle 및 저체온치료병합요법군, melatonin 및 저체온치료의 병합요법군으로 나누었다. Melatonin은 30 mg/kg 용량으로 복강 내 주사하였는데, 저산소 허혈 손상 직후 및 24시간 경과 시점으로 구분해서 2차례 투여하였다. 저체온치료는 전신 냉각으로 27℃ 환경에서 3시간동안 시행하였다. 정상 대조군은 저산소 허혈 뇌손상을 유발하지 않은 sham 수술군으로 하였다. 수술 후 3일(P10), 1주 경과(P14) 및 4주 시점(P35)에서 실험쥐를 희생시켜서 뇌손상을 평가하였다. 결과: TTC 염색에서 vehicle 투여한 P10 실험쥐는 저산소 허혈로 인한 뇌경색이 대조군에 비해 현저하였다. 또한, H&E염색에서 P35 실험쥐는 손상측 뇌위축이 현저하였다. Melatonin이나 저체온치료 단독으로는 뇌의 경색이나 위축을 유의하게 억제하지 못하였으나, melatonin 및 저체온요법의 병합치료는 뇌손상을 줄이는 효과가 있었다. In situ zymography 및 TUNEL 염색, 면역형광법을 이용해서 뇌손상을 평가한 결과 오직 병합요법만이 신경 보호효과를 나타내었다. 결론: Melatonin은 저산소 허혈 뇌손상에 대하여 저체온치료의 신경 보호작용에 상승효과를 가지는 것으로 보인다.

Melatonin이 암세포주의 미세구조 및 세포바깥기질단백에 미치는 영향

성언기(Eon-Ki Sung),정현규(Hyeon-Gyoo Jeong),송인환(In-Hwan Song),김주영(Joo-Young Kim),이융창(Yungchang Lee) 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.2

암세포의 증식을 억제하고 분화를 유도하는 melatonin을 0.2mM과 2mM의 농도로 α-MEM 배양액에 첨가하여 HeLa, HepG2, A549, L929 및 NIH/3T3 세포들을 배양하였다. 이들 세포주에서 암세포들의 전이와 밀접한 관계가 있는 당단백, fibronectin, laminin 및 actin의 양적인 변화에 미치는 melatonin의 영향을 PAS 또는 PAP 염색으로 광학현미경하에서 조사하였고, 미세구조에 어떤 변화를 일으키는지 투과전자현미경으로 관찰하여 세포소기관 수준에서 이 약제의 항암작용 영향력을 검토하였다. Melatonin을 2mM의 농도로 처리하면 2일, 3일에 세포의 증식이 억제되었다. Melatonin을 0.2mM과 2mM의 농도로 1일, 2일 및 3일 처리하면 모든 세포주에서 당단백, fibronectin, laminin 및 actin의 양이 대조군에 비해 대부분 증가하였다. 약제처리 후 2일, 3일에 모든 세포의 0.2mM과 2mM군에서 대조군에 비해 핵 내 이질염색질이 덩어리를 형성하였다. HeLa와 L929 세포의 경우 1일, 2일 및 3일에 0.2mM과 2mM군에서 과립형질내세망과 골지복합체의 수가 증가하였다. HeLa, A549 및 L929 세포에서 3일에 0.2mM과 2mM군에서 용해소체의 수적인 증가가 일어났다. 모든 세포에서 1일, 2일 및 3일에 0.2mM과 2mM군에서 소포들이 증가되었다. 이상의 성적을 종합해 볼 때 melatonin은 2mM의 농도로 적어도 2일간 처리한 후에야 항유사분열효과가 기대되었다. Melatonin을 0.2mM과 2mM의 농도로 처리하면 fibronectin과 laminin의 생성을 증가시켜 이것들이 결국 세포바깥으로 분비되어 세포바깥기질단백의 양을 증가시킬 것이고 이들 세포바깥기질단백과 형질막에 있는 integrin을 통해 연결된 actin의 증가는 세포의 부착성을 더욱 높이는 것으로 생각되었다. 또한 암세포가 전이되기 위해서는 세포바깥기질단백들을 뚫고 나가는데 필요한 여러 효소를 분비해야 하는데 세포바깥기질단백의 증가로 더욱 어려워질 것으로 생각되었다. Melatonin을 0.2mM과 2mM의 농도로 처리하였을 때 나타나는 핵내 이질염색질의 덩어리와 소포들의 증가와 같은 형태변화들은 melatonin의 작용을 평가하는 지표로 삼을 수 있을 것으로 생각되었다. Melatonin could be used as an anticancer agent to suppress the proliferation of tumor cells and induce the differentiation of cancer cells. HeLa, HepG2, A549, L929, and NIH/3T3 cell lines were cultivated in α-MEM with 0.2mM/2 mM melatonin. The influences of melatonin on quantitative changes of glycoprotein, fibronectin, laminin and actin related to the metastasis of tumor cells investigated with PAS or PAP at light microscopic level. To elucidate the possibility of antitumor actions of melatonin, the changes of cell organelles were observed under transmission electron microscope. Cell proliferation was suppressed after treatment with 2 mM melatonin for 2 or 3 days. Compared with control groups, the amounts of glycoprotein, fibronectin, laminin and actin in all cell lines at 1st, 2nd and 3rd day after treatment with 0.2 mM and 2 mM melatonin were generally increased. Heterochromatin in the nucleus formed clumps in all cell lines at 2nd and 3rd day after treatment with 0.2 mM and 2 mM melatonin. The numerical increase of rough endoplasmic reticulum and golgi complex observed in HeLa and L929 cells treated with 0.2 mM and 2 mM melatonin at 1st, 2nd and 3rd day. The number of lysosomes increased in HeLa, A549, and L929 cells treated with 0.2 mM and 2mM melatonin at 3rd day. The number of vesicles increased in all cell lines treated with 0.2mM and 2mM melatonin at 1st, 2nd and 3rd day. Taken together, antimitotic effect of melatonin can be expected at least 2day after treatment with 2 mM melatonin. The production of fibronectin and laminin in all cell lines treated with 0.2mM or 2mM melatonin increased. Therefore, the increase of amounts of extracellular matrix proteins in the extracellular space can be expected. And the increase of amounts of actin connected to the extracellular matrix proteins through the integrin of plasma membrane seemed to strengthen cell attachment. In order to metastasize of cancer cells, it is important for them to secrete various enzymes to pass through the extracellular matrix proteins. Hence, it will be more difficult for the cells to metastasize into other regions due to the increase of the extracellular matrix proteins. It was postulated that the clumps of heterochromatin and the numerical increase of vesicles induced by treatment with 0.2 mM and 2 mM melatonin could be represented for the actions of melatonin as morpholgical criteria.

Meta-analysis of the effect of melatonin application on abiotic stress tolerance in plants

Yang Xiaoxiao,Ren Jianhong,Li Juanjuan,Lin Xinyue,Xia Xiangyu,Yan Wenjie,Zhang Yuxin,Deng Xiping,Ke Qingbo 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.1

Melatonin is a hormone-like substance that promotes plant growth and development and alleviates stress levels. Although the physiological roles of melatonin and the underlying mechanisms have been qualitatively reviewed in plants, we do not fully understand when and how to apply melatonin to maximize its benefits. Here, we performed a meta-analysis to quantitatively evaluate the effect of melatonin on abiotic stress tolerance in plants and to determine the number of parameters modulated by melatonin. Melatonin significantly alleviated the growth inhibition induced by drought stress compared with other abiotic stresses, including salt, cold, heat, nitrogen deficit, and heavy metal toxicity, mainly owing to higher photosynthesis efficiency and antioxidant enzyme activity. Furthermore, melatonin modulated plant growth in a concentration-dependent manner and was more effective when applied to plants under moderate drought stress at an early stage via root irrigation. In addition, the impact of melatonin was greater in monocots than in dicots. Moreover, endogenous melatonin levels could be significantly increased via transgenic strategies. Among melatonin biosynthesis-related gene members, ASMT has tended to have the most influence on melatonin content in plants. In light of the rapidly developing genome editing technology, quantitatively increasing endogenous melatonin level in plant would be quite useful for moderating climatic conditions and combating desertification. Taken together, our results provide guidelines for melatonin application in crops plants for improving productivity under ongoing climate change.

Melatonin이 암세포주의 미세구조 및 세표바깥기질단백에 미치는 영향

성언기,정현규,김주영,1이융창,송인환 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.2

Isoproterenol과 Melatonin의 첨가가 돼지 단위발생란의 체외발달에 미치는 영향

정다운,윤윤진,박희성 한국수정란이식학회 2016 한국동물생명공학회지 Vol.31 No.1

In this study, to improve the in vitro development of various cells including cloned embryos, the effects that isoproterenol and melatonin have on in vitro development of porcine parthenogenetic oocytes were investigated. Parthenogenetic activation was induced with electrical stimulation, BSA and 6-DMAP treatment. 10-7 M of melatonin and isoproterenol (10-10, 10-12 and 10-14 M) were supplemented for in vitro maturation (IVM) and in vitro culture (IVC) medium, with different concentrations. When isoproterenol and melatonin were supplemented in IVM medium with different concentrations, there was no significant (P<0.05) difference of maturation rate in the treatment groups as well as in that of only melatonin. As isoproterenol and melatonin were supplemented in IVM medium with different concentrations, blastocyst rates of isoproterenol 10-12 M treatment group (37.1%) were significantly (P<0.05) higher than control group (26.0%). Isoproterenol and melatonin were supplemented in IVC medium with different concentrations, then the cleavage rate of 10-12 M isoproterenol treatment group (82.2%) was significantly (P<0.05) higher than the group that melatonin was only supplemented (70.9%). There was no difference of blastocyst rate between the treatment groups. When isoproterenol and melatonin were supplemented for IVM+IVC medium with different concentrations, the cleavage rate of 10-12 M isoproterenol treatment group (92.5%) was significantly (P<0.05) higher than the control group (82.8%) and the group that melatonin was only treated (81.6%). The blastocyst rate of 10-12 M as 45.6% was significantly (P<0.05) higher than control group (25.2%) and melatonin treatment group (31.2%). The cell number of blastocyst in 10-12 M isoproterenol treatment group 35.5±3.4 was significantly (P<0.05) highest. The results of this study showed that the development rate of IVC when both isoproterenol and melatonin were supplemented was higher than when melatonin was only supplemented. Therefore, it is concluded that isoproterenol is rather effective in the activation of melatonin. 10-7 M melatonin and 10-12 M isoproterenol were considered suitable concentration.

Melatonin alleviates asphyxial cardiac arrest-induced cerebellar Purkinje cell death by attenuation of oxidative stress

Cho, Jeong Hwi,Tae, Hyun-Jin,Kim, In-Shik,Song, Minah,Kim, Hyunjung,Lee, Tae-Kyeong,Kim, Young-Myeong,Ryoo, Sungwoo,Kim, Dae Won,Lee, Choong-Hyun,Hwang, In Koo,Yan, Bing Chun,Kang, Il Jun,Won, Moo-Ho Academic Press 2019 Experimental neurology Vol.320 No.-

<P><B>Abstract</B></P> <P>Although multiple reports using animal models have confirmed that melatonin appears to promote neuroprotective effects following ischemia/reperfusion-induced brain injury, the relationship between its protective effects and activation of autophagy in Purkinje cells following asphyxial cardiac arrest and cardiopulmonary resuscitation (CA/CPR) remains unclear. Rats used in this study were randomly assigned to 6 groups as follows; vehicle-treated sham operated group, vehicle-treated asphyxial CA/CPR operated group, melatonin-treated sham operated group, melatonin-treated asphyxial CA/CPR operated group, PDOT (a MT2 melatonin receptor antagonist) plus (+) melatonin-treated sham operated group and PDOT+melatonin-treated asphyxial CA/CPR operated group. Melatonin (20 mg/kg, i.p., 4 times before CA and 3 times after CA) treatment significantly improved survival rate and neurological deficit compared with the vehicle-treated asphyxial CA/CPR rats (survival rates ≥40% vs 10%), showing that melatonin treatment exhibited protective effect against asphyxial CA/CPR-induced Purkinje cell death. The protective effect of melatonin against CA/CPR-induced Purkinje cell death paralleled a remarkable attenuation of autophagy-like processes (Beclin-1, Atg7 and LC3), as well as a dramatic reduction in superoxide anion radical (O<SUB>2</SUB>·-), intense enhancements of CuZn superoxide dismutase (SOD1) and MnSOD (SOD2) expressions. Furthermore, the protective effect was notably reversed by treatment with PDOT, which is a selective MT2 antagonist. In brief, melatonin conferred neuroprotection against asphyxial CA/CPR-induced Purkinje cell death via inhibiting autophagic activation by reducing expressions of O<SUB>2</SUB>·- and increasing expressions of antioxidant enzymes, and suggests that MT2 is involved in neuroprotective effect of melatonin against Purkinje cell death caused by asphyxial CA/CPR.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Melatonin prevents cerebellar Purkinje cell death following asphyxial cardiac arrest. </LI> <LI> Melatonin attenuates autophagy-like processes via MT2 receptor. </LI> <LI> Melatonin attenuates increase of superoxide anion radical via MT2 receptor. </LI> <LI> Melatonin maintains antioxidant enzymes expressions via MT2 receptor. </LI> </UL> </P>

흰쥐 뇌줄기에서 광주기 및 melatonin주입이 serotonin면역반응에 미치는 영향

김명순,전승원,이창현 대한해부학회 2003 Anatomy & Cell Biology Vol.36 No.6