• 제목/요약/키워드: real-time qPCR

검색결과 311건 처리시간 0.024초

Four Novel Synthetic Tryptamine Analogs Induce Head-Twitch Responses and Increase 5-HTR2a in the Prefrontal Cortex in Mice

  • Abiero, Arvie;Ryu, In Soo;Botanas, Chrislean Jun;Custodio, Raly James Perez;Sayson, Leandro Val;Kim, Mikyung;Lee, Hyun Jun;Kim, Hee Jin;Seo, Joung-Wook;Cho, Min Chang;Lee, Kun Won;Yoo, Sung Yeun;Jang, Choon-Gon;Lee, Yong Sup;Cheong, Jae Hoon
    • Biomolecules & Therapeutics
    • /
    • 제28권1호
    • /
    • pp.83-91
    • /
    • 2020
  • Tryptamines are monoamine alkaloids with hallucinogenic properties and are widely abused worldwide. To hasten the regulations of novel substances and predict their abuse potential, we designed and synthesized four novel synthetic tryptamine analogs: Pyrrolidino tryptamine hydrochloride (PYT HCl), Piperidino tryptamine hydrochloride (PIT HCl), N,N-dibutyl tryptamine hydrochloride (DBT HCl), and 2-Methyl tryptamine hydrochloride (2-MT HCl). Then, we evaluated their rewarding and reinforcing effects using the conditioned place preference (CPP) and self-administration (SA) paradigms. We conducted an open field test (OFT) to determine the effects of the novel compounds on locomotor activity. A head-twitch response (HTR) was also performed to characterize their hallucinogenic properties. Lastly, we examined the effects of the compounds on 5-HTR1a and 5-HTR2a in the prefrontal cortex using a quantitative real-time polymerase chain reaction (qRT-PCR) assay. None of the compounds induced CPP in mice or initiated SA in rats. PYT HCl and PIT HCl reduced the locomotor activity and elevated the 5-HTR1a mRNA levels in mice. Acute and repeated treatment with the novel tryptamines elicited HTR in mice. Furthermore, a drug challenge involving a 7-day abstinence from drug use produced higher HTR than acute and repeated treatments. Both the acute treatment and drug challenge increased the 5-HTR2a mRNA levels. Ketanserin blocked the induced HTR. Taken together, the findings suggest that PYT HCl, PIT HCl, DBT HCl, and 2-MT HCl produce hallucinogenic effects via 5-HTR2a stimulation, but may have low abuse potential.

Spatial and seasonal distributions of the phototrophic dinoflagellate Biecheleriopsis adriatica (Suessiaceae) in Korea: quantification using qPCR

  • Kang, Hee Chang;Jeong, Hae Jin;Ok, Jin Hee;You, Ji Hyun;Jang, Se Hyeon;Lee, Sung Yeon;Lee, Kyung Ha;Park, Jae Yeon;Rho, Jung-Rae
    • ALGAE
    • /
    • 제34권2호
    • /
    • pp.111-126
    • /
    • 2019
  • The phototrophic dinoflagellate Biecheleriopsis adriatica is a small suessioid species characterized by a fragile thin wall. Although the morphology of this dinoflagellate is well established, there is currently little information available on its distribution and the environmental factors that influence this distribution. Thus, to investigate the spatial and seasonal distributions of the vegetative cells of B. adriatica in Korean waters, surface water samples were collected on a seasonal basis from 28 stations in the East, West, and South Sea of Korea and Jeju Island from April 2015 to October 2018, and abundances of the vegetative cells of B. adriatica were quantified using quantitative real-time polymerase chain reactions, for which we developed the species-specific primer and probe set. Simultaneously, major environmental parameters, including temperature, salinity, nutrient concentrations, and dissolved oxygen concentrations were measured. The vegetative cells of B. adriatica were detected at 20 of the 28 sampling stations: 19 stations in summer and 6 in autumn, although from no stations in either spring or winter. The ranges of water temperature and salinity at sites where this species was detected were $17.7-26.4^{\circ}C$ and 9.9-34.3, respectively, whereas those of nitrate and phosphate concentrations were not detectable-96.2 and $0.18-2.66{\mu}M$, respectively. Thus, the sites at which this species is found are characterized by a narrow range of temperature, but wide ranges of salinity and concentrations of nitrate and phosphate. The highest abundance of the vegetative cells of B. adriatica was $41.7cells\;mL^{-1}$, which was recorded in Jinhae Bay in July 2018. In Jinhae Bay, the abundance of vegetative cells was significantly positively correlated with the concentration of nitrate, but was negatively correlated with salinity. On the basis of these findings, it appears that the abundance of B. adriatica vegetative cells shows strong seasonality, and in Jinhae Bay, could be affected by the concentrations of nitrate.

계대 배양 속도가 다른 과잉치 치수유래 줄기세포 간 유전자 발현 특성 (Gene Expression of Supernumerary Dental Pulp Related to the Subculture Speed: A Pilot Study)

  • 이유경;김종수;신지선;김종빈
    • 대한소아치과학회지
    • /
    • 제46권2호
    • /
    • pp.219-225
    • /
    • 2019
  • 이 연구의 목적은 과잉치 치수 유래 줄기세포의 계대 배양 속도에 대한 상아모세포 연관 유전자의 발현을 비교하는 것이다. 줄기세포는 다른 여러 형태의 세포로 분화할 수 있는 미 분화된 세포이다. 이는 환경이나 특정 자극에 의해 세포 분열이 일어나며 근육이나 골 같은 특정 장기의 조직으로 분화할 수 있다. 20명의 어린이에서 발거한 과잉치에서 과잉치 치수 유래 줄기세포가 얻어졌다. 10계대까지 배양하는 동안 가장 빠른 속도로 계대 배양된 세포와 가장 느린 속도로 계대 배양된 세포 각 3계대와 10계대 세포를 얻어 실험을 진행하였다. 각 세포는 분화제를 처리한 군과 처리하지 않은 군으로 나누었다. 이 실험에서 발현도를 살펴본 유전자는 Osteonectin (ONT), Osteocalcin (OCN), Alkaline Phosphatase (ALP), Dentin matrix acidic phosphoprotein 1 (DMP-1), Dentin sialophosphoprotein (DSPP)이다. 분화가 된 세포가 전반적으로 더 높은 유전자 발현도를 보였으며, 미분화 세포는 10계대에서, 분화된 세포는 3계대에서 더 높은 유전자 발현도를 보였다. 빠른 계대 배양 속도를 보인 세포가 OCN과 DSPP를 제외하고 상대적으로 더 낮은 유전자 발현도를 보였다.

Comparison of the spatial-temporal distributions of the heterotrophic dinoflagellates Gyrodinium dominans, G. jinhaense, and G. moestrupii in Korean coastal waters

  • Lee, Sung Yeon;Jeong, Hae Jin;Kang, Hee Chang;Ok, Jin Hee;You, Ji Hyun;Park, Sang Ah;Eom, Se Hee
    • ALGAE
    • /
    • 제36권1호
    • /
    • pp.37-50
    • /
    • 2021
  • Heterotrophic dinoflagellates Gyrodinium spp. are one of the major grazers of phytoplankton in many coastal waters. Gyrodinium dominans, G. jinhaense, and G. moestrupii have similar morphologies but different edible prey species. To explore the variations in the ecological niches of these three species, we investigated their spatial-temporal distributions in Korean waters. Because of the high similarity in morphology among these three Gyrodinium species, we used real-time polymerase chain reactions to quantify their abundance in water samples that were seasonally collected from 28 stations along the Korean Peninsula from April 2015 to October 2018. Cells of G. dominans were found at all sampling stations, G. jinhaense at 26 stations, and G. moestrupii at 22 stations, indicating that all three species were widely distributed in Korea. Furthermore, all three species displayed strong seasonal distributions. The largest numbers of the stations where G. dominans and G. jinhaense cells were present were found during the summer (26 and 23 stations, respectively), but that for G. moestrupii was found in the autumn (15 stations). The abundance of G. dominans was positively correlated with that of G. jinhaense, but not with that of G. moestrupii. The highest abundances of G. dominans (202.5 cells mL-1) and G. jinhaense (20.2 cells mL-1) were much greater than that of G. moestrupii (1.2 cells mL-1). The highest abundances of G. dominans and G. jinhaense were found in July, whereas that of G. moestrupii was found in March. The abundances of G. dominans and G. jinhaense, but not G. moestrupii, were positively correlated with water temperature. Therefore, the spatial-temporal distributions of G. dominans and G. jinhaense were closer than those of G. moestrupii and G. dominans or G. jinhaense. This differs from results based on the relative differences in ribosomal DNA sequences and the types of edible prey reported in the literature. Thus, the variations in spatial-temporal distributions and prey species of these three Gyrodinium species suggest that they may have different ecological niches in Korean coastal waters.

염생식물인 갯끈풀과 갯메꽃 추출물의 HaCaT 세포에서 피부 보습 및 피부 장벽 기능에 미치는 영향 (Effect of Halophyte (Spartina anglica and Calystegia soldanella) Extracts on Skin Moisturizing and Barrier Function in HaCaT Cells)

  • 하유나;정재우;이원휘;오준혁;김연정
    • 한국해양생명과학회지
    • /
    • 제6권2호
    • /
    • pp.58-65
    • /
    • 2021
  • 노화가 진행될수록 활성산소종으로 인하여 피부 보습은 떨어지고 피부 장벽은 붕괴되어 피부가 손상된다. 본 연구에서는 인천 동막 해변에 서식하는 염생식물인 갯끈풀(Spartina anglica; SAE)과 갯메꽃(Calystegia soldanella; CSE)을 70% 에탄올(EtOH)로 추출하여 피부 보습 및 피부 장벽 기능 강화에 대한 효능을 평가하였다. 이 추출물들에 대한 피부 각질형성세포(HaCaT cell)에서 세포독성을 WST-8 assay를 이용하여, 세포 생존율이 90% 이상을 보이는 농도를 선별하여 추가 실험을 진행하였다. ABTS 라디칼 소거능을 통해 항산화 효과를 확인한 결과, SAE와 CSE는 높은 라디칼 소거능을 보였다. 피부 보습과 관련된 인자들인 filaggrin (FGL), aquaporin 3(AQP3), hyaluronan synthase 2 (HAS2)과 피부 장벽 기능과 연관 있는 transglutaminase 1 (TGM1)과 involucrin (INV)의 유전자 수준에서의 발현 변화를 측정한 결과, SAE에 의해 AQP3, HAS2, TGM1의 발현이 증가하였으나, CSE는 변화가 없는 것을 확인할 수 있었다. SAE에 의한 세포 내 신호전달 경로를 확인하기 위해 western blot 분석을 수행하였다. Extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen activated protein kinase의 활성이 SAE에 의하여 상향 조절되었음을 확인하였다. 이러한 결과는 갯끈풀 추출물이 피부 보습 및 피부 장벽 기능 강화를 위한 화장품의 기능성 소재로 사용될 수 있음을 시사한다.

Ginsenoside Rg1 alleviates Aβ deposition by inhibiting NADPH oxidase 2 activation in APP/PS1 mice

  • Zhang, Han;Su, Yong;Sun, Zhenghao;Chen, Ming;Han, Yuli;Li, Yan;Dong, Xianan;Ding, Shixin;Fang, Zhirui;Li, Weiping;Li, Weizu
    • Journal of Ginseng Research
    • /
    • 제45권6호
    • /
    • pp.665-675
    • /
    • 2021
  • Background: Ginsenoside Rg1 (Rg1), an active ingredient in ginseng, may be a potential agent for the treatment of Alzheimer's disease (AD). However, the protective effect of Rg1 on neurodegeneration in AD and its mechanism of action are still incompletely understood. Methods: Wild type (WT) and APP/PS1 AD mice, from 6 to 9 months old, were used in the experiment. The open field test (OFT) and Morris water maze (MWM) were used to detect behavioral changes. Neuronal damage was assessed by hematoxylin and eosin (H&E) and Nissl staining. Immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction (q-PCR) were used to examine postsynaptic density 95 (PSD95) expression, amyloid beta (Aβ) deposition, Tau and phosphorylated Tau (p-Tau) expression, reactive oxygen species (ROS) production, and NAPDH oxidase 2 (NOX2) expression. Results: Rg1 treatment for 12 weeks significantly ameliorated cognitive impairments and neuronal damage and decreased the p-Tau level, amyloid precursor protein (APP) expression, and Aβ generation in APP/PS1 mice. Meanwhile, Rg1 treatment significantly decreased the ROS level and NOX2 expression in the hippocampus and cortex of APP/PS1 mice. Conclusions: Rg1 alleviates cognitive impairments, neuronal damage, and reduce Aβ deposition by inhibiting NOX2 activation in APP/PS1 mice.

향부자 에틸아세테이트 분획물의 염증 및 가려움증에 관한 연구 (A Study on Inflammation and Itching of Cyperus rotundus Ethyl Acetate Fractions)

  • 김도규;이지연;문소현;김유경;김나리;정아름;장준환;이재섭;한상배;배준태
    • 대한화장품학회지
    • /
    • 제48권4호
    • /
    • pp.303-312
    • /
    • 2022
  • 본 연구에서는 향부자를 이용하여 에틸아세테이트로 분획물을 제조하고, 지표물질 α-cyperone의 함량을 확인하였다. 또한, 향부자 에틸아세테이트 분획물(EAFC) 및 α-cyperone의 항염증 및 가려움증 완화 효능평가를 통하여 기능성 소재로서의 가능성을 확인하였다. EAFC 내의 α-cyperone의 함량을 확인하기 위해 HPLC 분석을 실시한 결과, α-cyperone의 함량이 5.243%임을 확인하였다. EAFC 및 지표성분인 α-cyperone의 염증 완화 효과를 검증하기 위해 lipopolysaccharide (LPS)로 염증반응을 유도시킨 RAW 264.7 대식세포에서의 nitric oxide (NO) 생성 저해능을 확인한 결과, 농도의존적으로 NO 생성을 저해하였다. 추가적인 염증염증 유발 인자인 IL-1β, IL-6, TNF-α, COX-2 및 iNOS의 mRNA 발현 역시 농도의존적으로 저해하였다. EAFC를 함유한 간단한 화장품 제형을 이용하여 임상시험을 실시한 결과, sodium lauryl sulfate (SLS)로 유발된 피부 자극이 진정되는 효과 및 가려움증이 완화되는 효과를 확인하였다. 이를 통하여 향부자는 염증 저해 및 가려움증 완화 효과를 가지는 천연 화장품 원료로 사용 가능할 것으로 사료된다.

Green perilla leaf extract ameliorates long-term oxidative stress induced by a high-fat diet in aging mice

  • Edward, Olivet Chiamaka;Thomas, Shalom Sara;Cha, Kyung-Ok;Jung, Hyun-Ah;Han, Anna;Cha, Youn-Soo
    • Nutrition Research and Practice
    • /
    • 제16권5호
    • /
    • pp.549-564
    • /
    • 2022
  • BACKGROUND/OBJECTIVES: Oxidative stress is caused by an imbalance between harmful free radicals and antioxidants. Long-term oxidative stress can lead to an "exhausted" status of antioxidant defense system triggering development of metabolic syndrome and chronic inflammation. Green perilla (Perilla frutescens) is commonly used in Asian cuisines and traditional medicine in southeast Asia. Green perilla possesses numerous beneficial effects including anti-inflammatory and antioxidant functions. To investigate the potentials of green perilla leaf extract (PE) on oxidative stress, we induced oxidative stress by high-fat diet (HFD) in aging mice. MATERIALS/METHODS: C57BL/6J male mice were fed HFD continuously for 53 weeks. Then, mice were divided into three groups for 12 weeks: a normal diet fed reference group (NDcon), high-fat diet fed group (HDcon), and high-fat diet PE treated group (HDPE, 400 mg/kg of body weight). Biochemical analyses of serum and liver tissues were performed to assess metabolic and inflammatory damage and oxidative status. Hepatic gene expression of oxidative stress and inflammation related enzymes were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: PE improved hepatopathology. PE also improved the lipid profiles and antioxidant enzymes, including hepatic glutathione peroxidase (GPx) and superoxide dismutase (SOD) and catalase (CAT) in serum and liver. Hepatic gene expressions of antioxidant and anti-inflammatory related enzymes, such as SOD-1, CAT, interleukin 4 (IL-4) and nuclear factor erythroid 2-related factor (Nrf2) were significantly enhanced by PE. PE also reduced the levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in the serum and liver; moreover, PE suppressed hepatic gene expression involved in pro-inflammatory response; Cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6). CONCLUSIONS: This research opens opportunities for further investigations of PE as a functional food and possible anti-aging agent due to its attenuative effects against oxidative stress, resulting from HFD and aging in the future.

p66Shc in sheep preimplantation embryos: Expression and regulation of oxidative stress through the manganese superoxide dismutase-reactive oxygen species metabolic pathway

  • Tong Zhang;Jiaxin Zhang;Ruilan Li
    • Animal Bioscience
    • /
    • 제36권7호
    • /
    • pp.1022-1033
    • /
    • 2023
  • Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

Identification and functional prediction of long non-coding RNAs related to oxidative stress in the jejunum of piglets

  • Jinbao Li;Jianmin Zhang;Xinlin Jin;Shiyin Li;Yingbin Du;Yongqing Zeng;Jin Wang;Wei Chen
    • Animal Bioscience
    • /
    • 제37권2호
    • /
    • pp.193-202
    • /
    • 2024
  • Objective: Oxidative stress (OS) is a pathological process arising from the excessive production of free radicals in the body. It has the potential to alter animal gene expression and cause damage to the jejunum. However, there have been few reports of changes in the expression of long noncoding RNAs (lncRNAs) in the jejunum in piglets under OS. The purpose of this research was to examine how lncRNAs in piglet jejunum change under OS. Methods: The abdominal cavities of piglets were injected with diquat (DQ) to produce OS. Raw reads were downloaded from the SRA database. RNA-seq was utilized to study the expression of lncRNAs in piglets under OS. Additionally, six randomly selected lncRNAs were verified using quantitative real-time polymerase chain reaction (qRT-PCR) to examine the mechanism of oxidative damage. Results: A total of 79 lncRNAs were differentially expressed (DE) in the treatment group compared to the negative control group. The target genes of DE lncRNAs were enriched in gene ontology (GO) terms and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathways. Chemical carcinogenesis-reactive oxygen species, the Foxo signaling pathway, colorectal cancer, and the AMPK signaling pathway were all linked to OS. Conclusion: Our results demonstrated that DQ-induced OS causes differential expression of lncRNAs, laying the groundwork for future research into the processes involved in the jejunum's response to OS.