• 제목/요약/키워드: human ALDH2

검색결과 16건 처리시간 0.021초

Expression of Human Mitochondiral Aldehyde Dehydrogenase 2 in Mammalian Cells using Vaccinia Virus-T7 RNA Polymerase

  • Kang, Su-Min;Yoo, Seung-Ku;Lee, Ki-Hwan
    • Journal of Microbiology
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    • 제37권1호
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    • pp.41-44
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    • 1999
  • Human mitochondrial aldehyde dehydrogenase 2 (ALDH2) is mainly responsible for oxidation of acetaldehyde generated during alcohol oxidation in vivo. A full-length cDNA of human liver ALDH2 was successfully expressed using a vaccinia virus-T7 RNA polymerase system. The expressed ALDH2 had an enzymatic activity as high as the native human liver ALDH2 enzyme.

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대장균에서 사람 ALDH2 유전자의 발현 (Expression of Human ALDH2 Gene in escherichia coli)

  • 곽보연;이기환;정한승
    • 한국식품영양학회지
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    • 제10권2호
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    • pp.268-271
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    • 1997
  • 사람의 미토콘드리아에 있는 aldehyde dehydrogenase(ALDH2)는 체내에서 알코올 대사 과정 중에 생성되는 아세트알데히드를 산화시키는 주된 역할을 담당하고 있다. 이 ALDH2가 알코올 대사에 미치는 영향을 연구하기 위하여 가용화된 효소가 필요하다. 알려져 있는 유전자의 염기서열 데이터를 바탕으로 ALDH2의 cDNA는 cDNA 라이브러리에서 선별하였으며, 이를 여러 가지 대장균 발현벡터에 연결하였다. 제조한 발현벡터를 형질전환시킨 대장균을 사용하여 단백질의 발현을 확인한 결과 대부분의 계에서 ALDH가 과발현되고 있었다. 그러나 발현된 단백질의 대부분은 inclusion body로 형성되어, 실제로 가용화된 효소의 양은 전체 발현된 양의 5% 이하 였고 이들 몇 가지 발현 system으로 재조합 미오2DML 발현을 확인하였다.

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필치가 혈중 알코올농도와 간세포내 알코올대사 효소 활성에 미치는 영향 (Effect of Feelch on Blood Alcohol Concentration in Human and Hepatic Alcohol Metabolizing Enzyme Activity in Alcohol-Fed Rats)

  • 이오미;예충민;최병철;이지윤;강혜정;최윤경;김창종;심상수
    • 약학회지
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    • 제49권4호
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    • pp.340-346
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    • 2005
  • To investigate the effect of Feelch on alcohol metabolism, we measured both blood alcohol concentration in human and hepatic alcohol metabolizing enzyme activity in rats. The blood alcohol concentration in Feelch-ingested group was significantly lower than that in water-ingested group at 0, 40, and 80 minute after alcohol intake. The blood alcohol concentration between male and female taken 300ml of $21\%$ alcohol showed the significant differences; the peak value of blood alcohol concentration in male and female were $0.083\pm0.014\%\;and\;0.108\pm0.018\%$, respectively. In alcohol-fed rats, aldehyde dehydrogenase (ALDH) activity was significantly increased, whereas alcohol dehydrogenase (ADH) activity was not changed. In both Feelch-fed group and Feelch plus alcohol-fed group, ADH and ALDH activity were significantly increased as compared with each control group. Feelch decreased phospholipase $A_2$ activity and lipid peroxidation in hepatic tissue and activities of serum aminotransferases as compared with control. These results suggest that Feelch may have a hepatoprotective effects and this is likely due to lower blood alcohol concentration via the increment of hepatic ADH and ALDH activity.

Lactobacillus brevis HY7401 섭취가 쥐의 혈중 알코올 수준에 미치는 영향 (Effect of Lactobacillus brevis HY7401 Intake on the Serum Ethanol Concentration in Rats)

  • 안영태;김용희;배진성;임광세;허철성;양우영;김형수;백영진
    • 한국식품과학회지
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    • 제36권4호
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    • pp.604-608
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    • 2004
  • Lactobacillus 균주들의 에탄올과 acetaldehyde의 대사 가능성을 시험관내 및 생체내에서 시험하였다. ADH와 ALDH 활성은 L. brevis 균주들이 다른 Lactobacillus 균주들과 비교하여 높게 나타났으며, 특히 L. brevis HY7401은 가장 높은 ADH와 ALDH 활성을 나타냈다. L. brevis HY7401$(10^9\;cfu/mL)$을 2주간 하루에 1 mL씩 쥐에게 급여한 후 시험 14일째에 에탄올(4g/kg BW)을 경구투여하고 혈중 에탄올 양을 측정한 결과, L. brevis HY7401 급여에 의해 혈중 에탄올 수준이 대조구에 비해 유의적으로 감소하는 것으로 나타났다. 에탄올 투여 6시간후의 혈중 에탄올 양이 각각 $1,195{\pm}92.9$(대조군) 그리고 $454{\pm}150mg/L$(HY7401군)으로 나타났다. 또한 L. brevis HY7401 급여한 군에서 소장내 에탄올 농도가 대조구와 비교하여 약 1/2수준이었으며, 초산 농도는 2배정도 높은 것으로 확인되었다. 따라서 이 연구결과들은 사람으로부터 분리된 L. brevis HY7401이 높은 ADH와 ALDH 활성을 갖고 있어 에탄올과 acetaldehyde를 대사한다는 것을 의미하고 있다. 또한 L. brevis HY7401을 쥐에게 급여함으로써 혈중 에탄올 수준을 낮출 수 있는 것으로 확인되었다.

Effect of Lactobacillus fermentum MG590 on Alcohol Metabolism and Liver Function in Rats

  • Kim, Ji-Hyun;Kim, Hyun-Jin;Son, Jeong-Hwa;Chun, Ho-Nam;Yang, Jin-Oh;Choi, Sung-Jin;Paek, Nam-Soo;Choi, Gyoung-Hoon;Kim, Sung-Koo
    • Journal of Microbiology and Biotechnology
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    • 제13권6호
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    • pp.919-925
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    • 2003
  • Alcohol consumption has numerous health consequences for the human body. For example, heavy drinking on a daily basis causes liver diseases, and certain products such as acetaldehyde produced from alcohol metabolism are more toxic than alcohol itself. Accordingly, the current study evaluated the role of Lactobacillus fermentum MG590 to enhance the removal of the toxic effect of alcohol in alcohol metabolism. The maximum activities of the alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) by L. fermentum MG590 were observed after 6 h of culture. The production of ADH and ALDH by L. fermentum MG590 was also confirmed by SDS-PAGE. Six hours after the addition of alcohol to a culture broth of L. fermentum MG590, the alcohol concentration decreased from 7.5 to 2.7%. From an in vitro evaluation based on hepatocytes, the viability of hepatocytes in a medium containing alcohol and the cytosol of L. fermentum MG590 was higher than that in a medium containing only alcohol. From an in vivo test using SD rats fed a 22% alcoholic drink, the blood alcohol concentration (BAC), glutamic-oxaloacetic transaminase (GOT), and glutamic-pyruvic transaminase (GPT) in the rats fed a medium containing L. fermentum MG590 were lower than those in the rats fed a medium containing only the alcohol drink. These results demonstrate that the ADH and ALDH produced by L. fermentum MG590 play an important role in detoxicating alcohol in vivo. Therefore, a fermentation broth of L. fermentum MG590 could be used as an effective alcohol detoxification drink.

황기(黃芪)의 재배 년수에 따른 면역 및 항산화 활성 연구 (Studies on Immunomodulatory and Antioxidant Activities of Astragali membranacei Radix according to the Cultivated Years)

  • 정철
    • 대한한방피부미용학회지
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    • 제1권1호
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    • pp.53-90
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    • 2005
  • Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

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흰쥐에서 황칠나무 열수 추출물을 포함한 혼합물의 혈중 알코올 농도와 숙취 해소 효과 (Eliminatory Effect of Mixture including Hot Water Extract of Dendropanax morbifera Lev. on Alcohol-induced Blood Alcohol Concentration and Hangover in Rat)

  • 나주련;김은;박소이;이기훈;정의선;김진석;김용재;김선오
    • 한국키틴키토산학회지
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    • 제23권4호
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    • pp.267-276
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    • 2018
  • 본 연구에서는 숙취해소에 좋은 것으로 알려진 식품 소재와 황칠나무 추출물을 복합하여 in vitro 및 in vivo에서 알코올 유도된 간 손상에 대한 보호효과를 검토하였다. HepG2세포에서 300 mM 알코올과 SBJ 혼합물을 처리하였을 때 농도 의존적으로 LDH 수치가 감소하였으나 황칠나무 추출물을 제외한 복합물인 SBJ-복합물에서는 그 효과가 유의적으로 낮아지는 것을 관찰하였다. In vivo에서 SBJ 혼합물의 보호효과를 확인하고자 흰쥐에 알코올과 SBJ 혼합물을 투여한 결과, 알코올 투여 후 1시간까지 급격하게 혈중 알코올 수치가 EtOH군에서 증가하는 것이 관찰되었으며, SBJ 혼합물 투여군은 유의적인 차이로 감소되었음이 관찰되었으며 또한 농도의존적인 경향을 확인하였다. ADH 및 ALDH 활성의 증가는 SBJ 혼합물의 알코올 분해 및 대사산물의 제거 활성에 기여할 것으로 예상된다. 뿐만 아니라, 알코올에 의하여 증가한 LDH의 농도가 대조군과 유사한 수준으로 유지하는 것을 확인하였으며, GST, SOD, GPx 및 reduced glutathione와 같은 항산화 인자 및 효소의 활성은 대조군보다 EtOH군이 유의적으로 감소했으며, SBJ 혼합물에 의해 개선되는 경향을 관찰하였다. 이를 통해 SBJ 혼합물은 ADH, ALDH 활성 증가 및 항산화 방어계를 향상시킴으로써 산화적 스트레스 감소를 통한 간보호 효과가 있는 것으로 생각되며, 이러한 간보호에 미치는 주요한 추출물은 황칠나무 추출물임을 알 수 있었다. 이러한 결과를 토대로 향후 숙취해소 작용을 갖는 신규 식품소재로 활용될 수 있을 것으로 기대된다.

Two novel mutations in ALDH18A1 and SPG11 genes found by whole-exome sequencing in spastic paraplegia disease patients in Iran

  • Komachali, Sajad Rafiee;Siahpoosh, Zakieh;Salehi, Mansoor
    • Genomics & Informatics
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    • 제20권3호
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    • pp.30.1-30.9
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    • 2022
  • Hereditary spastic paraplegia is a not common inherited neurological disorder with heterogeneous clinical expressions. ALDH18A1 (located on 10q24.1) gene-related spastic paraplegias (SPG9A and SPG9B) are rare metabolic disorders caused by dominant and recessive mutations that have been found recently. Autosomal recessive hereditary spastic paraplegia is a common and clinical type of familial spastic paraplegia linked to the SPG11 locus (locates on 15q21.1). There are different symptoms of spastic paraplegia, such as muscle atrophy, moderate mental retardation, short stature, balance problem, and lower limb weakness. Our first proband involves a 45 years old man and our second proband involves a 20 years old woman both are affected by spastic paraplegia disease. Genomic DNA was extracted from the peripheral blood of the patients, their parents, and their siblings using a filter-based methodology and quantified and used for molecular analysis and sequencing. Sequencing libraries were generated using Agilent SureSelect Human All ExonV7 kit, and the qualified libraries are fed into NovaSeq 6000 Illumina sequencers. Sanger sequencing was performed by an ABI prism 3730 sequencer. Here, for the first time, we report two cases, the first one which contains likely pathogenic NM_002860: c.475C>T: p.R159X mutation of the ALDH18A1 and the second one has likely pathogenic NM_001160227.2: c.5454dupA: p.Glu1819Argfs Ter11 mutation of the SPG11 gene and also was identified by the whole-exome sequencing and confirmed by Sanger sequencing. Our aim with this study was to confirm that these two novel variants are direct causes of spastic paraplegia.

Establishment of a Pancreatic Cancer Stem Cell Model Using the SW1990 Human Pancreatic Cancer Cell Line in Nude Mice

  • Pan, Yan;Gao, Song;Hua, Yong-Qiang;Liu, Lu-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권2호
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    • pp.437-442
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    • 2015
  • Aim: To establish a pancreatic cancer stem cell model using human pancreatic cancer cells in nude mice to provide a platform for pancreatic cancer stem cell research. Materials and Methods: To establish pancreatic cancer xenografts using human pancreatic cancer cell line SW1990, nude mice were randomly divided into control and gemcitabine groups. When the tumor grew to a volume of $125mm^3$, they treated with gemcitabine at a dose of 50mg/kg by intraperitoneal injection of 0.2ml in the gemcitabine group, while the mice in control group were treated with the same volume of normal saline. Gemcitabine was given 2 times a week for 3 times. When the model was established, the proliferation of pancreatic cancer stem cells was observed by clone formation assay, and the protein and/or mRNA expression of pancreatic stem cell surface markers including CD24, CD44, CD133, ALDH, transcription factors containing Oct-4, Sox-2, Nanog and Gli, the key nuclear transcription factor in Sonic Hedgehog signaling pathway was detected by Western blot and/or RT-PCR to verify the reliability of this model. Results: This model is feasible and safe. During the establishment, no mice died and the weight of nude mice maintained above 16.5g. The clone forming ability in gemcitabine group was stronger than that of the control group (p<0.01). In gemcitabine group, the protein expression of pancreatic cancer stem cell surface markers including CD44, and ALDH was up-regulated, the protein and mRNA expression of nuclear transcription factor including Oct-4, Sox-2 and Nanog was also significantly increased (P<0.01). In addition, the protein expression of key nuclear transcription factor in Sonic Hedgehog signaling pathway, Gli-1, was significantly enhanced (p<0.01). Conclusions: The pancreatic cancer stem cell model was successfully established using human pancreatic cancer cell line SW1990 in nude mice. Gemcitabine could enrich pancreatic cancer stem cells, simultaneously accompanied by the activation of Sonic Hedgehog signaling pathway.

Artesunate와 비스테로이드소염제 NSAID의 병용 처리에 의한 항암 활성 증강 (Potentiation of Anticancer Effect of Artesunate by Combination with Nonsteroidal Anti-inflammatory Drugs on Human Cancer Cells)

  • 문현정;강치덕;김선희
    • 생명과학회지
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    • 제31권10호
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    • pp.873-884
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    • 2021
  • 본 연구에서는 다양한 인체 암포주를 대상으로 NSAID의 항암 효과를 증강시키는 artesunate (ART)의 역할과 이에 대한 분자적 기전을 연구하였다. 다양한 타입의 암세포주를 대상으로 암세포 성장 억제 활성을 조사한 결과, ART는 NSAID인 celecoxib (CCB) 또는 dimethyl-CCB (DMC)와의 병용 효과를 나타내었다. ART 처리로 ATF4/CHOP의 발현 증강과 함께 오토파지 유도 표식인 p62 감소의 결과로서, ATF4/CHOP 경로가 ART의 오토파지 유도 활성에 관여할 것으로 예상되었으며, ART의 오토파지 활성과 관련하여 NRF2 및 암 줄기 세포 관련 단백질인 CD44, CD133, ALDH1, Oct4, mutated p53 (mutp53) 및 c-Myc의 발현이 감소되었다. 또한 DMC 단독처리 보다 ART와 DMC의 병용으로 ATF4/CHOP의 발현 증강과 p62의 감소가 더욱 촉진되고, NRF2 및 암 줄기 세포 관련 단백질 발현 감소도 현저히 촉진되면서 궁극적으로 PARP 활성화에 의해 apoptosis가 유도됨을 알 수 있었다. 이러한 결과는 ART/DMC 병용 처리가 각 물질 단독 처리보다 암세포의 성장 억제 및 apoptosis 유도에 더욱 효과적이고, ART 및 DMC 의 오토파지 유도 활성은 암 줄기 세포 관련 단백질의 분해를 촉진함으로써, 암 줄기 세포가 제거될 수 있음을 시사하였다. 이와 같이 ART는 NSAID 뿐만 아니라 imatinib의 항암 효과를 증강시키는 활성으로, chemosensitizer로서 중요한 후보 물질이 될 수 있음을 밝혔다.