카페인 유도 자가포식이 프리온 매개 신경 세포 독성에 미치는 영향 및 기전 연구

문지홍 전북대학교 일반대학원 2014 국내석사

Prion diseases are neurodegenerative disorders characterized by the accumulation of an abnormal isoform of the prion protein PrPSc. The human prion protein fragment PrP (106-126) possesses a majority of the pathogenic properties associated with the infectious scrapie isoform of PrP, known as PrPSc. Accumulation of PrPSc in the brain of humans and animals affects the central nervous system. Recent epidemiologic studies suggest that caffeine, one of the major components of coffee, has a protective effect against the development of neurodegeneration; however, a protective function of caffeine in prion disease has not been reported. We therefore investigated the effect of caffeine on prion-mediated neurotoxicity. Expression of the autophagy marker LC3-II protein was dose-dependently increased by caffeine, and the autophagy induced by caffeine protected neuronal cells against PrP (106-126)-induced cell death. On the other hand, down-regulation of LC3-II with the autophagy blockers 3-methyladenine and wortmannin prevented the caffeine-mediated neuroprotective effects. This report provides the first evidence that treatment with caffeine protects against prion-mediated neurotoxicity and that this neuroprotection is mediated by caffeine-induced autophagy signals. We suggest that caffeine treatment may be a therapeutic strategy for prion peptide-induced apoptosis.

Effects of caffeine / MG132 on the vitrification of mouse mature oocyte and further embryonic development

백지이 차의과학대학교 대학원 2016 국내석사

Cryopreservation is the technique used to stop all biological activities by cooling the cells to low temperature (-196 ℃) for future use. This is important to remove the intracellular water of the oocyte by dehydration using short exposure to high concentrations of cryoprotectants (Fuller et al, 1996). Most importantly, it could reduce the damage to oocytes without the forming of ice crystals Cryopreservation has been developed to store surplus embryos after Assisted Reproductive Technology (ART). Recently, vitrification is replaced with slow freezing protocol, because of improved survival rates and clinical outcomes. Vitrification using a high concentration of CPAs and it makes the cell toxicity increased. Also risk of contamination increases due to plugged into liquid nitrogen directly. Ice crystal formation, bubble formation could happen during vitrification and warming process and may reduce the survival and developmental rates (Fiamigni et alL, 2004). Generally, Cyclin B1 interacts with Cdk1 to form a complex known as the MPF, which is plays an essential role in the cell cycle regulation and maintaining the meiotic arrest of oocytes (Pines, 1999; Labbe et al, 1989). Succu (2011) reported that, MPF activity has been observed to decrease in MII ovine oocytes after vitrification, which could compromise the oocyte development after in vitro fertilization. Likewise, MAPK also responsible for the beginning of germinal vesicle breakdown and is essential for the maintenance of the meiotic arrest of oocytes at MII-stage (Dupre et al, 2002; Araki et al, 1996). The present study were designed to investigate the effects of the Caffeine and MG132 of vitrified/warmed MII mouse oocytes on the activity of MPF and MAPK. We evaluated their survival, fertilization, cleavage and developmental rates. Ovulated MII oocytes were retrieved from 6 weeks old B6D2F1 female mice at 14 hr post hCG injections. Collected MII oocytes were vitrified/warmed into divided 4 groups (non-treated, 10 mM caffeine, 10 μM MG13, 10 mM caffeine + 10 μM MG132). After warming, MPF and MAPK activity was evaluated using ELISA and Western blotting. Also, verified the fertilization and blastocyst rates of mature oocytes after in vitro fertilization. Blastocyst formation was increased in 10 mM caffeine, 10 μM MG132 and 10 mM caffeine + 10 μM MG132. Also MPF activity were same tendency. However, statistically no difference was found all groups in MAPK activity and differential staining. These results suggest that during vitrification/warming process treatment of caffeine and MG132 can prolongs the MPF activity and stabilized the chromatin configuration, it helps these fertilization and blastocyst rates. In conclusion, caffeine and MG132 may reduce Cryoinjury during vitrification/warming process, and furthermore, these caffeine and MG132 could be useful for the development and supplementation of cryopreservation method.

Drug interaction of a propenone compound with caffeine in rats : 랫드에서 프로페논 화합물과

엄연지 영남대학교 일반대학원 2015 국내석사

Caffeine을 연속투여한 흰쥐에 있어 Adenosine 및 ATP에 의한 심혈관계의 반응성변화

李炅姬 同德女子大學校 大學院 1995 국내석사

Cardiovascular responses of adenosine or ATP were evaluated in caffeine treated rat comparing to the normal rat. Caffeine was administered for 6 weeks in drinking water as a concentration of 300mg/ℓ. Bolus intravenous injection of caffeine showed the increase of blood pressure and heart rate in the anesthetized rats, and the increase of contractile force and heart rate in isolated spontaneously beating right atrium of the rats. But there was no significant difference on the caffeine induced cardiovascular responses between normal rats and caffeine treated rats. Bolus intravenous injection of adenosine or ATP showed dose-dependent decrease of blood pressure and heart rate in the anesthetized rats. The effect of adenosine or ATP on the heart rate was significantly inhibited in chronic caffeine treated rat comparing to the normal rat. But the change of blood pressure was not. Adenosine or ATP showed dose-dependent decrease of contractile force and heart rate in isolated spontaneously beating right atrium of therats. These effects of adenosine were significantly increased in chronic caffeine treated rat comparing to the normal rat. But the effects of ATP were not. Adenosine and ATP were respectively showed endotheliumindependent and endothelium-dependent vasorelaxation in the isolated thoracic aorta. These effects of adenosine were significantly increased in chronic caffeine treated rat comparing to the normal rat. But the effects of ATP were not. On the above results, the responses of adenosine on the heart and blood vessel were increased in chronic caffeine-treated rat comparing to the normal rat. But the effects of ATP were not. These results were due to the up-regulation of adenosine receptor in chronic caffeine treated rats. And these results might be suggested that the incidence of cardiovascular diseases including myocardiac infarction and arrythmia would be decreased in chronic caffeine treated rats.

Caffein이 Rhodotorula glutinis의 지질대사에 미치는 영향

최경식 건국대학교 대학원 1992 국내석사

Caffeine이 Yeast의 생육 및 지방 대사에 미치는 영향을 조사하고자 caffeine을 0%, 0.01%, 0.05%, 0.10% 첨가한 Glucose 배지에 Rhodotorula glutinis를 접종하여 8일간 진탕 배양하면서 지질 생성량, 증식도, pH, 당 소비율, ATP : citrate layse의 활성을 조사하였다. 1. 증식도는 caffeine 첨가량 증가순으로 감소되어 0.10% 첨가구가 가장 낮았다. 2. pH는 배양 1일째 모든 시험구에서 급격한 변화가 있었으며 배양 기간 동안 점차 낮아지는 경향을 보였으나, 0.10% 첨가구는 다른 시험구보다 높은 pH를 보였다. 3. 당 소비율은 모든 시험구가 대조구보다 작았으나 0.01%와 0.05% 첨가구는 비슷하였으며, 0.10% 첨가구는 대조구보다 현저히 낮았다. 4. ATP : citrate lyase의 활성은 첨가구 모두 배양 4일째까지 대조구에 비해 낮았으나, 0.01% 첨가구의 5일째 활성은 대조구의 최고 활성보다 1.2배 증가하였다. 5. 균체 지질 함량은 대조구에 비해 0.01% 첨가구가 1.1배로 증가하였으나, 0.05% 첨가구는 0.76배의 작은 함량을 보였다. 6. Caffeine 0.01% 첨가구는 생육도와 당 소비율은 대조구에 비해 낮았으나, 효소 활성 및 지질 생성량은 높았다. This study was carried out to investigate the effect of caffeine on the lipid contents, growth rate, pH, sugar consuiption rate and ATP:citrate lyase activity of Rhodotorula glutinis. 1. Cell growth of Rhodotorula glutinis decreased in the media containing 0.10% caffeine than that of control group at 26℃ for 8 days incubation. 2. pH change was remarkably decreased in early incubation time and continuously decreased in all incubation time, but 0.10% caffeine containing group, it was higher than other groups. 3. Sugar consumption rate decreased in the order of addition of caffeine. 4. ATP: Citrate lyase activity decreased until 4 days incubation time but the activity of 0.01% caffeine containing group was increased 1.2 times than that of control group at 5 days incubation respectively. 5. Lipid contents of 0.01% caffeine containing group increased 1.1 times than that of control group respectively. 6. Cell growth and sugar consumption rate of 0.0l% caffeine containing group decreased but lipid content are increased compared with that of control group.

Caffeine의 藥物動態에 미치는 Histamine H2 拮抗劑의 影響(I)

강인호 우석대학교 1987 국내석사

본 실험자는 cimetidine, ranitidine 전투여가 caffrine의 흡수, 분포, 대사, 배설 등에 영향을 미칠 수 있다고 사려되어 소장흡수, 혈중농도, 뇨중농도, 위내산도, 수면시간 등을 측정한 결과 몇가지 지견을 얻었기에 보고하는 바이다. Cimetidine, ranitidine 전투여가 caffeine의 흡수, 분포, 대사, 배설 등에 영향을 줄 수 있는 것으로 생각되어 소장흡수실험, 혈중농도, 뇨중농도 비교실험, 위내산도 실험, 수면시간에 미치는 영향등을 측정한 결과 아래와 같은 결론을 얻었다. 1. Caffeine의 장내 흡수율은 caffeine단독 투여군에 비하여 cimetidine, ranitidine 전투여한 cimetidine이 높았다. 특히 ranitidine전투여시 caffeine의 흡수율이 cimetidine전투여시 보다 높았다. 2. Caffeine의 혈중농도는 caffeine 단독 투여군에 비하여 cimetidine, ranitidine 전투여군이 높았으며 cimetidine 전투여군은 caffeine의 혈중 반감기를 다소 연장시킴으로써 체내 유지시간을 연장시켰다. 3. Cimetidine 전투여시 cimetidine은 caffeine의 뇨중 배설을 억제시켰으나 ranitidine 전투여시 ranitidine은 caffeine의 배설에 별 영향을 미치지 못하였다. 4. 전투여된 cimetidine과 ranitidine은 caffeine의 위산분비작용을 억제 하였고 그 작용은 ranitidine이 cimetidine보다 상당히 강하였다. 5. 전투여된 cimetidine과 ranitidine은 caffeine의 수면 단축작용을 상승시켜 수면시간이 상당히 단축되었고 그 작용은 cimetidine 전투여가 ranitidine 전투여 보다 강하였다. 이상의 결과로 보아 caffeine제제가 치료를 위한 약제로써 투여 될 때 위·십이지장 궤양환자의 cimetidine,ranitidine 투여량이 고려 되었으면 한다. 특히 cimetidine이 투여되는 환자에 있어서는 caffeine 함량을 줄이거나 cimetidine 제제 대신 ranitidine 제제로 대치 투여하는 방법 등의 투여방법이 고려 되었으면 한다.

Caffeine이 Saccharomyces cerevisiae의 生理와 酵素活性에 미치는 影響

김요승 建國大學校 大學院 1990 국내석사

Caffeine이 Saccharomyces cerevisiae의 生理에 미치는 影響을 조사하기 위하여 포도당 기본배지에 caffeine을 0, 0.001, 0.01, 0.1, 0.5 % (w/v) 씩 添加하고 S. cerevisiae ATCC 4124를 접종하여 72시간 정치배양(30℃)하면서 增殖率, alcohol含量, pH, 糖消費率, 醱酵率등의 變化와 정치배양한 효모균체의 ADH 및 PyDC 浩性을 調査하였다. 1. S. cerevisiae ATCC 4124의 增殖은 전 발효기간중 0.001% 첨가구는 대조구보다 증가하였고 0.01 %, 0.1% 첨가구는 억제되었으며, 0.5 % 첨가구는 대조구에 비해 현저하게 억제되었다. 2. pH의 變化는 caffeine의 添加量이 많을수록 변화폭이 적었으며 48시간 이후에 완만하게 감소하였다. 3. 各 試驗區의 alcohol 含量은 발효 48시간에서 0.001 %구(3.77%)> 대조구 (3.20%)> 0.01%구 (3.18%)> 0.1%구 (3.08%)>0.5%구(1.71 %) 순으로 0.001 % caffeine 첨가구의 alcohol 含量이 대조구에 비하여 1.17배 높았고 0.01 %, 0.1 %, 0.5 % 첨가구의 함량은 대조구의 함량보다 각각 1.0배, 1.04배 낮았으며 특히 0.5 % caffeine 첨가구의 alcohol 함량은 대조구보다 1.87배 낮았다. 4. ADH 活性은 0.001% (S_(1))구> 0%(S_(0))구> 0.01%(S_(2))구>0.1 %(S_(3))구>0.5%(S_(4))구 순으로 0.001 % caffeine 첨가구의 ADH 활성은 대조구보다 1.28배 높았으며 0.01, 0.1, 0.5 % 첨가구의 ADH 활성은 대조구보다 각각 1.32배, 2.13배, 3.35배 낮았다. 또한 PyDC 活性도 0.001% (S_(1))구> 0%(S_(0))구> 0.01%(S_(2))구>0.1%(S_(3))구> 0.5%(S_(4))구 순으로 0.001 % caffeine 첨가구의 PyDC활성은 대조구의 활성에 비해 각각 1.66배, 3.27배, 5.42배 감소하였다. This study was carried out to investigate the effect of caffeine on growth, the rate of alcohol fermentation, and the enzyme activity of alcohol dehydrogenase and pyruvate decarboxylase of Sacchar-omyces cerevisiae. S. cerevisiae was inoculated into the glucose solution media containing 0, 0.001, 0.01, 0.1 and 0.5% of caffeine respectively and which was fermented at 30℃ for 72 hours. 1. The cell growth of S. cerevisiae was more increased in the media containing 0.001% caffeine than in the control group at 30℃ for 72 hour fermentation. But the growth was inhibited in the medium containing 0.01, 0.1, and 0.5% caffeine. Especially the growth in the medium containing 0.5% caffeine was remarkably inhibited than control group. 2. pH change was remarkably decreased in early fermentation time. But slowly decreased after 24 hour fermentation. 3. After 48 hour fermentation, alcohol content was increased in the order of· 0.001%(3.77%)) control group(3.20%)> 0.01%(3.18%) > 0.1%(3.08%)> 0.5% (1.71%) group. The alcohol content of 0.001% group was increased to 1.17 times that of control group, but 0.01%1 0.1%, 0.5% group was decreased to 1.00, 1.04, 1.87 times that of control group. 4. Alcohol dehydrogenase(ADH) and pyruvate decarboxy-lase(PyDC) activity was increased in the order of 0.001%> control group> 0.01%> 0.1% >0.5% group. The ADH activity in 0.001% group was increased to 1.28 times that of control group. But the ADH activity in 0.01%, 0.1%, 0.5% group was decreased to 1.32, 2.13, 3.35 times that of control group, respectively. The PyDC activity in 0.001% group was increased to 1.11 times that of control group. But the PyDC activity in 0.01%1 0.1%, 0.5% group was decreased to 1.66, 3.12, 5.42 times that of control group, respectively.

Caffeine triggers calcium signaling and confers resistance against a broad-spectrum of biotic/abiotic stresses in rice

박종찬 경희대학교 생명공학원 2018 국내박사

작물은 수확량을 심각하게 감소시키는 생물학적/비 생물학적 스트레스를 비롯한 환경 요인에 지속적으로 노출된다. 이러한 스트레스에 대처하기 위해 화학 농약이 일반적으로 사용되었지만, 과도한 적용으로 인하여 종종 환경 문제가 발생한다. 따라서 기존의 방법보다 안전하고 효과적인대체 방법을 찾는 것이 시급하다. 여기에서 나는 카페인의 형질 전환접근법을 통하여 벼의 기본 면역을 높인다는 것을 보고한다. 카페인 (1,3,7-trimethylxabthine)은 퓨린 알카로이드 계열의 전형적인 2 차 대사산물중의 하나이며 커피, 카카오, 차와 같은 몇 가지 식물에서 합성된다. 카페인은 병원균과 초식동물에 독성을 일으켜 숙주 식물을 보호하는 것으로 알려져 있다. 카페인은 purine catabolism pathway에서 xanthosine 을 전구체로 사용하여 합성된다. Coffea arabica 에서 동정 된3 개의 N-메틸 전이 효소 유전자를 도입하여 카페인 생성벼(CPR)을만들었다. CPR 은 표현형 적으로 원형벼와 동일했다. CPR 종자의 크기와 무게, 또한 생산량은 원형 벼와 비교하여 거의 차이가 없었다. 카페인의합성은 벼에서 생산되는 전구체, xanthosine의 양에 민감하게 좌우되었다.CPR 의 xanthosine과 카페인의 합성양은 8 배엽 단계에서 상관적으로 크게 증가하였으며 Xoo 와 M. oryzae에 대한 완전한 저항력을 갖추었다. 또한 CPR 은 벼 멸구 뿐만 아니라 가뭄 및 염스트레스에 대한 저항성이 강화되었다. 나는 카페인이 궁극적으로 살리실산의 합성을 활성화시키는 일련의 신호 전달 작용을 시작하는 분자 경로를 확인하였다. 이 결과는 방어 프라이밍 (defence priming)이라 불리는 병에 대한 대처법의 새로운 형태를 통하여 생물학적 및 비 생물학적 스트레스에 높은 수준의 내성을부여/제공하는 효과적인 수단임을 시사한다. 이는 벼뿐만 아니라 다른 작물 품종에 대한 장래의 분자육종에 잠재적으로 가치 있는 방어프라이밍을 위한 새로운 전략이 될 수 있다. Crop plants are constantly exposed to environmental factors, including biotic / abiotic stresses, which severely reduce yields. To cope with such stresses, chemical pesticides have commonly been used, but their excessive application has often caused environmental problems. It is thus urgent to find an alternative ways that are safer and more effective than conventional procedures. Here I report that caffeine elevates the basic immunity in rice through transgenic approach. Caffeine (1, 3, 7-trimethylxanthine) is a typical secondary metabolite of purine alkaloids and is synthesized in a few plant species such as coffee, cacao, and tea. Caffeine is thought to protect host plants by its toxicity against pathogens and herbivores. Caffeine is synthesized using a precursor, as the xanthosine in purine catabolism pathway. A caffeine-producing rice (CPR) was then generated by introduction of three N-methyltransferase genes identified from Coffea arabica. CPR plants were phenotypically equal to a wild type. The size and weight of the CPRs seed, also yields almost no difference as compared to the wild type. Caffeine synthesis sensitively depends on amount of precursor, xanthosine produced in rice. Xanthosine and caffeine amounts in CPR are correlatively increased at eight-leaf stages and the CPR has full resistance to Xoo and M. oryzae, at eight stages. In addition, CPR plants showed enhanced resistance not only to white backed plant hoppers but also to drought and salt stresses. The molecular pathway was identified through which caffeine initiates a series of signal cascade that ultimately activates synthesis of salicylic acid. These results suggest that caffeine is an effective means of conferring / provide a high level of resistance to biotic and abiotic stresses through a new form of disease coping method called defense priming. This can be a novel strategy for defense priming, being potentially valuable for future molecular breeding of not only rice but also other crop varieties.

Regulation of germ cell development by caffeine intake in Caenorhabditis elegans : Caffeine intake increases germ cell apoptosis by modulating vitellogenesis and mitochondrial function

김주해 건국대학교 대학원 2024 국내석사

Caffeine (1,3,7-trimethylxanthine) is a widely consumed bioactive substance worldwide. A recent study showed that caffeine intake reduced vitellogenesis and the number of progeny. Interestingly, the reduction in both reproduction and vitellogenesis caused by caffeine intake was improved by vitamin B12 supplementation, which is an essential co-factor in methionine metabolism. Caffeine intake-induced impairment of methionine metabolism decreased vitellogenesis and increased germ cell apoptosis in a LIN-35/RB-dependent manner, through mitochondrial-dependent pathway. Furthermore, the effect of methionine metabolism on the caffeine-ingested C. elegans was assessed. It was examined that caffeine intake decreased essential enzymes related to the methionine cycle and this alteration of methionine metabolism induced a reduction of vitellogenesis and the increased germ cell apoptosis. The increased germ cell apoptosis and autophagic activity were restored to normal levels by methionine supplementation through increasing vitellogenesis in the caffeine-ingested C. elegans. These results indicate that methionine supplementation plays a beneficial role in the germ cell health and the offspring development by regulating vitellogenesis and mitochondrial activity. 카페인(1,3,7- trimethylxanthine)은 전 세계적으로 널리 소비되는 생리활성 물질입니다. 최근 연구에 따르면 카페인 섭취는 난황 단백질의 생성 과 자손의 수를 감소시켰습니다. 흥미롭게도, 메티오닌 대사에 필수적인 보조 요소인 비타민 B12 보충이 카페인 섭취로 인해 감소한 난황 단백질의 생성량과 생식이 개선되었다는 것이 보고되었습니다. 따라서 본 연구에서는 메티오닌 대사의 역할이 카페인을 섭취한 선충의 생식에 미치는 영향을 조사했습니다. 카페인 섭취로 인한 메티오닌 대사 손상은 난황 단백질 생성량의 감소를 일으키고, LIN-35/RB 경로에 의존적인 생식세포 사멸을 증가시켰으며, 이는 미토콘드리아 기능의 변화에 의한 경로를 통해 이루어질 수 있습니다. 다음으로 메티오닌 대사가 카페인 섭취한 C. elegans에 미치는 영향을 확인하였습니다. 카페인 섭취는 메티오닌 주기와 관련된 필수 효소를 감소시키며, 이 메티오닌 대사의 변화는 난황 단백질 생성량의 감소와 생식 세포 사멸 증가를 유발했습니다. 메티오닌 보충은 카페인 섭취한 C. elegans에서 생식 세포 사멸과 자가 포식 활성을 정상 수준으로 회복시켰습니다. 이 결과들은 메티오닌 보충이 난황 단백질 생성과 미토콘드리아 활동을 조절하여 생식 세포 건강과 후손 발달에 유익한 역할을 한다는 것을 나타냅니다.

Caffeine 수면억제에 대한 감초 (Glycyrrhizae Radix) 의 완화효과

전태동 경희대학교 대학원 2018 국내박사

이 연구는 감초 추출물이 Caffeine에 의해 유발된 Sprague Dawley Dawley (SD) rat의 수면장애에 미치는 영향을 평가하기 위해 시행되었다. 실험은 세포내 실험과 동물실험의 2가지로 이루어 졌으며, 실험은 Caffeine 투여군, Caffeine과 감초를 동시에 투여한 군으로 나누어서 이루어 졌으며, 세포내 실험은 SD rat의 해마 단면으로부터 발생하는 신경활동 신호를 multielectrode array (MEA) 를 이용하여 측정하였고, 동물심험은 SD rat의 수면시간을 측정하였다. 세포내 실험에서는 Caffeine만 투여한 군에서는 처리 후 및 washing 후에 유의성 있는 변화가 나타났으며, Caffeine과 감초를 동시에 투여한 군에서는 감초 100μg/ml 을 투여한 군에서만 유의성 있는 변화가 나타났으며, Caffeine 군과 Caffeine과 감초 투여 군의 비교에서는 처치 직후에는 감초 400μg/ml 을 처리한 군에서만 유의성 있는 차이가 있었고, washing 후에는 감초 200μg/ml, 400μg/ml 을 처리한 군에서 유의성 있는 차이를 나타내었다. 감초와 Caffeine의 level 별 spike 수의 변화에서는 Caffeine 처리 군은 low level (21.63μV 이하) 과 high level 모두에서 처치 및 washing 후에 유의성 있는 변화가 나타났으며, Caffeine과 감초를 동시에 투여한 군에서는 유의성 있는 변화가 나타나지 않았다. 동물실험에서도 Caffeine을 투여한 군은 Control 군에 비해 유의성 있게 수면시간이 감소하였고, Caffeine과 감초를 동시에 투여한 군은 Caffeine 군과 비교하여 유의성 있게 수면시간을 증가시켰다. 결과적으로, 감초 추출물은 Caffeine의 부작용을 완화시키는데 효율적이며, Caffeine으로 인한 수면장애에 사용할 수 있는 새로운 물질이 될 수 있을 것으로 보인다. 앞으로 감초를 이용하여 Caffeine의 부작용을 완화시키는데 대한 추가적인 연구가 필요할 것으로 사료된다. This study was performed to evaluate the effects of Glycyrrhizae Radix (GR) water extract on sleep disturbance induced by caffeine in rats. The experiment was conducted in two ways; an ex-vivo experiment and in-vivo experiment. As for an ex-vivo experiment, the experimental groups was divided into 2 groups; the group treated with caffeine and both caffeine and GR. An ex-vivo experiment used organotypic hippocampal slice cultures in SD rats. Neurological signals were examined with the multi-channel extracellular recording system. Moreover, as for an in-vivo experiment, the experimental group was divided into 3 groups; the group treated with normal water, water with caffeine and water with both caffeine and GR. In an in-vivo experiment, each group's sleep duration was measured using the camera. As a result, there were a significant difference between the spike numbers(%) before and after caffeine treatment and there were not statistically significant difference among the groups treated with caffeine and GR at 200μg/mL and 400μg/mL of GR compared to before treatment, and those treated with 100μg/mL GR was significantly different from those before treatment. The group treated with both caffeine and 400μg/mL of GR showed the significant difference comparing to the group treated with caffeine. In addition, the group treated with caffeine and 200μg/mL as well as the group treated with caffeine and 400μg/mL showed the significant difference comparing to the group treated with caffeine after washing. In the caffeine treated group, the spike number of low level (21.63μV or less) and high level (21.63μV or more) were significantly increased after treatment and washing. There were no significant increase in caffeine and GR treatment groups. In the animal study, sleep duration of the group treated with caffeine was significantly decreased and it was recovered in rats treated with both caffeine and GR. In conclusion, the GR extracts had shown to be effective in controlling the side effects of caffeine, and as a result they seemed to be a new competing substance against sleep disturbance. More studies in the future need to be performed to alleviate the side effects of caffeine.