• 제목/요약/키워드: Human T-cell

검색결과 1,398건 처리시간 0.026초

비파엽의 핵수용체 관련 신규 약물 작용점 연구 : 만성 질환 치료 가능성 (Eriobotrya japonica inhibits the transactivation of nuclear orphan receptor : the possibility to treat the chronic diseases)

  • 김민진;김영우;박선동
    • 대한한의학방제학회지
    • /
    • 제30권4호
    • /
    • pp.225-231
    • /
    • 2022
  • Objectives : Oxidative damage has a variety of mechanism in the human organs including brain and liver. The purpose of this study is to examine the prevention against cellular damage, and the inhibitory effect on lipogenesis of the water extract of Eriobotrya japonica (EJE). Methods : The effect of EJE on cell viability was assessed using the MTT assay. To investigate the mechanism of EJE's inhibition of lipogenesis, we analyzed the relevant proteins using immunoblot analysis. Results : Induction of T0901317, a LXR agonist, significantly increased SREBP-1c expression, which was blocked by the pretreatment of EJE in the dose-dependent manners. This beneficial herb also regulated the genes related with SREBP-1c such as the acetyl-CoA carboxylase. Conclusion : These results suggested that EJE might have a possibility of the treatment of chronic diseases as mediated with the inhibition of LXRα/SREBP-1c pathway.

A novel tetrapeptide for the treatment of hair loss identified in ginseng berry: in silico characterization and molecular docking with TGF-β2

  • Sung-Gyu Lee;Sang Moon Kang;Hyun Kang
    • Journal of Plant Biotechnology
    • /
    • 제49권4호
    • /
    • pp.316-324
    • /
    • 2022
  • Hair loss causes psychological stress due to its effect on appearance. Therefore, the global market for hair loss treatment products is rapidly growing. The present study demonstrated that ginseng berry-derived and sequence-modified peptides promoted the proliferation rate of dermal papilla (DP) cells and keratinocytes, in addition to having antioxidant properties. Moreover, the potential role of these ginseng berry peptides as TGF-β2 antagonists was confirmed through in silico computer docking. In addition to promoting the growth of ,the ginseng berry-derived peptides also promoted the proliferation of keratinocytes experimental Particularly, an unmodified ginseng berry-derived peptide (GB-1) and two peptides with sequence modifications (GB-2 and GB-3) decreased ROS generation and exhibited a protective effect on damaged HaCaT keratinocytes. Computer-aided peptide discovery was conducted to identify the potential interactions of important proteins with transforming growth factor-beta 2 (TGF-β2), a key protein that plays a crucial role in the human hair growth cycle. Our results demonstrated that MAGH, an amino acid sequence present in herbal supplements and plant-based natural compounds, can inhibit TGF-β2.

A simple and rapid method for detection of single nucleotide variants using tailed primer and HRM analysis

  • Hyeonguk Baek;Inchul, Choi
    • 한국동물생명공학회지
    • /
    • 제38권4호
    • /
    • pp.209-214
    • /
    • 2023
  • Background: Single nucleotide polymorphisms (SNPs) are widely used genetic markers with applications in human disease diagnostics, animal breeding, and evolutionary studies, but existing genotyping methods can be labor-intensive and costly. The aim of this study is to develop a simple and rapid method for identification of a single nucleotide change. Methods: A modified Polymerase Chain Reaction Amplification of Multiple Specific Alleles (PAMSA) and high resolution melt (HRM) analysis was performed to discriminate a bovine polymorphism in the NCAPG gene (rs109570900, 1326T > G). Results: The inclusion of tails in the primers enabled allele discrimination based on PCR product lengths, detected through agarose gel electrophoresis, successfully determining various genotypes, albeit with some time and labor intensity due to the use of relatively costly high-resolution agarose gels. Additionally, high-resolution melt (HRM) analysis with tailed primers effectively distinguished the GG genotype from the TT genotype in bovine muscle cell lines, offering a reliable way to distinguish SNP polymorphisms without the need for time-consuming AS-PCR. Conclusions: Our experiments demonstrated the importance of incorporating unique mismatched bases in the allele-specific primers to prevent cross-amplification by fragmented primers. This efficient and cost-effective method, as presented here, enables genotyping laboratories to analyze SNPs using standard real-time PCR.

Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene

  • Kim, Bae-Chul;Han, Rong-Xun;Kim, Myung-Yoon;Shin, Young-Min;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
    • /
    • 제33권2호
    • /
    • pp.107-111
    • /
    • 2009
  • The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

체세포에 있어서 Knock-in 벡터 상동영역 구조에 따른 Knock-in 효율 (Knock-in Efficiency Depending on Homologous Arm Structure of the Knock-in Vector in the Bovine Fibroblasts)

  • 김세은;박다솜;구덕본;강만종
    • Reproductive and Developmental Biology
    • /
    • 제41권1호
    • /
    • pp.7-16
    • /
    • 2017
  • The knock-in efficiency in the fibroblast is very important to produce transgenic domestic animal using nuclear transfer. In this research, we constructed three kinds of different knock-in vectors to study the efficiency of knock-in depending on structure of knock-in vector with different size of homologous arm on the ${\beta}-casein$ gene locus in the somatic cells; DT-A_cEndo Knock-in vector, DT-A_tEndo Knock-in vector I, and DT-A_tEndo Knock-in vector II. The knock-in vector consists of 4.8 kb or 1.06 kb of 5' arm region and 1.8 kb or 0.64 kb of 3' arm region, and neomycin resistance gene(neor) as a positive selection marker gene. The cEndo Knock-in vector had 4.8 kb and 1.8 kb homologous arm. The tEndo Knock-in vector I had 1.06 kb and 0.64 kb homologous arm and tEndo Knock-in vector II had 1.06 kb and 1.8 kb homologous arm. To express endostatin gene as transgene, the F2A sequence was fused to the 5' terminal of endostatin gene and inserted into exon 7 of the ${\beta}-casein$ gene. The knock-in vector and TALEN were introduced into the bovine fibroblast by electroporation. The knock-in efficiencies of cEndo, tEndo I, and tEndo II vector were 4.6%, 2.2% and 4.8%, respectively. These results indicated that size of 3' arm in the knock-in vector is important for TALEN-mediated homologous recombination in the fibroblast. In conclusion, our knock-in system may help to create transgenic dairy cattle expressing human endostatin protein via the endogenous expression system of the bovine ${\beta}-casein$ gene in the mammary gland.

3T3-L1 지방전구세포에서 합토글로빈에 의한 염증성 cytokine 발현 조절 (The Effect of Haptoglobin on Expression of Inflammatory Cytokines in 3T3-L1 Preadipocytes.)

  • 조진경;김남훈;오미경;박선주;김인숙
    • 생명과학회지
    • /
    • 제18권4호
    • /
    • pp.537-541
    • /
    • 2008
  • 백색지방조직(white adipose tissue)은 에너지 저장뿐만 아니라 다양한 adipokines을 분비하는 중요한 내분비 기관이다. 급성기반응 단백질로 알려져 있는 합토글로빈(haptoglobin)도 adipokine의 한 종류로서 지방세포에서 합성되고 분비된다. 그러나 adipokine으로서의 기능과 지방조직에서의 역할은 아직까지 규명되지 않았다. 본 연구에서는 3T3-L1 지방전구세포를 합토글로빈 유전자로 transfection 시켜 합토글로빈을 과발현하는 세포를 만들고 세포증식, 염증관련 인자들의 발현조절 및 단구세포의 유인성을 조사하였다. 그 결과, 합토글로빈은 3T3-L1 세포의 성장에는 별 영향을 미치지 않았으나 IL-6와 COX-2 발현을 저해하고 HO-1 합성을 증가하였다. 또한 THP-1 단구세포를 이용한 invasion assay에서는 합토글로빈이 단구세포의 이동을 저해하였다. 이러한 결과들은 합토글로빈이 지방조직에서 항염증 반응에 관여함을 시사한다. 만성적 염증상태(chronic low-grade inflammatory state)로 인식되고 있는 비만은 염증관련 인자들에 의한 인슐린저항성이 유도되는 바, 합토글로빈은 비만 관련 인슐린저항성을 방지하고 인슐린민감성을 향상시키는 데에도 기여할 것으로 생각된다.

Comparative Genomic Analysis of Pathogenic Factors of Pectobacterium Species Isolated in South Korea Using Whole-Genome Sequencing

  • Jee, Samnyu;Kang, In-Jeong;Bak, Gyeryeong;Kang, Sera;Lee, Jeongtae;Heu, Sunggi;Hwang, Ingyu
    • The Plant Pathology Journal
    • /
    • 제38권1호
    • /
    • pp.12-24
    • /
    • 2022
  • In this study, we conducted whole-genome sequencing with six species of Pectobacterium composed of seven strains, JR1.1, BP201601.1, JK2.1, HNP201719, MYP201603, PZ1, and HC, for the analysis of pathogenic factors associated with the genome of Pectobacterium. The genome sizes ranged from 4,724,337 bp to 5,208,618 bp, with the GC content ranging from 50.4% to 52.3%. The average nucleotide identity was 98% among the two Pectobacterium species and ranged from 88% to 96% among the remaining six species. A similar distribution was observed in the carbohydrate-active enzymes (CAZymes) class and extracellular plant cell wall degrading enzymes (PCWDEs). HC showed the highest number of enzymes in CAZymes and the lowest number in the extracellular PCWDEs. Six strains showed four subsets, and HC demonstrated three subsets, except hasDEF, in type I secretion system, while the type II secretion system of the seven strains was conserved. Components of human pathogens, such as Salmonella pathogenicity island 1 type type III secretion system (T3SS) and effectors, were identified in PZ1; T3SSa was not identified in HC. Two putative effectors, including hrpK, were identified in seven strains along with dspEF. We also identified 13 structural genes, six regulator genes, and five accessory genes in the type VI secretion system (T6SS) gene cluster of six Pectobacterium species, along with the loss of T6SS in PZ1. HC had two subsets, and JK2.1 had three subsets of T6SS. With the GxSxG motif, the phospholipase A gene did locate among tssID and duf4123 genes in the T6SSa cluster of all strains. Important domains were identified in the vgrG/paar islands, including duf4123, duf2235, vrr-nuc, and duf3396.

Cathepsin D의 발현이 비소세포 폐암의 예후에 미치는 영향 (Influence of Cathepsin D Expression on Prognosis in Non-Small Cell Lung Cancer)

  • 염형렬;명재일;임종철;김한균;이남훈;이대호;고향미;문종영;강헌석;류형선;김완;박창수;박경옥
    • Tuberculosis and Respiratory Diseases
    • /
    • 제49권1호
    • /
    • pp.60-71
    • /
    • 2000
  • 배경 및 목적 : Cathepsin D는 리소솜에 위치하는 단백분해효소로서 종양의 침윤, 전이, 증식에 관여할 것으로 생각되며, 이러한 작용을 통해 예후에도 중요한 역할을 할 것으로 추정된다. 폐암에서의 Cathepsin D의 예후인자로서의 역할은 아직 확립되지 않고 논란이 많은 실정이다. 이 연구의 목적은 비소세포폐암에서 Cathepsin D 발현의 예후적인 중요성을 알고자 하였다. 방법 : 비소세포 폐암환자 중 치료 목적으로 수술적 처치를 시행한 총 54명의 환자를 대상으로 하여 적출한 폐조직의 면역조직화학적 염색으로 Cathepsin D의 발현을 관찰하고 생존기간 및 TNM 병기와의 관계를 보았다. 결과 : 종양세포에서의 Cathepsin D의 발현은 총 54례 중 18례에서 관찰되어 33.3%의 발현율을 보였으나, 발현군과 비발현군 사이에 조직학적 분화도, 암의 크기. 영역 림프절 침범, 병리조직적 병기(surgical-pathologic stage, p-stage)는 통계적인 유의한 차이를 보이지 않았다. 간질세포에서는 29례(53.7%)에서 중둥도에서 다량(moderate to massive)의 Cathepsin D가 발현되는 것이 관찰되었고, 발현양상과 병리조직적 병기사이에 통계적으로 유의한 관련성이 있었으나(p=0.031), 각각의 조직학적 분화도, 암의 크기, 영역 림프절 침범과는 관계가 없었다. 종양세포와 세포에서의 Cathepsin D 의 발현은 생존율로 표현한 예후와의 유의한 관련성이 없었다. 예후와 관련된 변수를 사용한 다변량 분석결과 영역림프절 침범이 유일한 독립적 예후인자가 되었으며 Cathepsin D는 예후 인자로서의 의미는 없었다. 결론 : 비소세포폐암의 간질세포내 Cathepsin D 발현양상은 병리조직적 병기와 유의한 관련성을 나타내어 종양 진행과의 관련 가능성을 제사하였으나, 다른 임상병리인자들 및 예후와의 관련성은 없었다. 종양세포내에서의 Cathepsin D 발현은 병리조직적 병기를 포함한 임상병리 인자들 및 예후와 관계가 없었다.

  • PDF

육종용약침의 항암작용(抗癌作用) 및 면역효과(免疫效果)에 대(對)한 실험적(實驗的) 연구(硏究) (Study on the Anti-cancer, Anti-metastasis and Immune response improvement of Aqua-acupuncture with Cistanches Herba infusion solution)

  • 이은용;이병렬
    • Journal of Acupuncture Research
    • /
    • 제17권1호
    • /
    • pp.251-286
    • /
    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement of aqua-acupuncture with Cistanches Herba infusion solution, we used Cistanches Herba infusion solution(taken by water-alcohol method) put into Chung-wan(CV12) and Chok-Samni(S36) of BALB/c or C57BL/6 which are corresponding to human body. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, MST, ILS, changes in amount of WBC, RBC, PLT, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-5}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 2. The cytotoxicity about HT1080 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-8}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 3. The effect on expression of MMP-9 gene was decreased in all the sample groups, compared with control group. 4. The effect on the control-ability on the cancer cell proliferation showed cytotooicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$ diluent groups. 5. S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group that showed same number of the control group. 6. S-180 cancer cell line transplants in BALB/c mice showed high MST and ILS significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group showed low MST and ILS than control group. 7. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS compared with control group significantly in anti-metastasis test. 8. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of WBC and glucose, and decreased significantly in the amount of LDH, compared with control group. However, there's no significant increase or decrease in number of RBC, PLT, total protein and creatinine. 9. We couldn't find any significant relation in spleen weight of the sample group. 10. In pulmonary colony, sample group was decreased significantly, compared with control group. 11. Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 12. In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 13. The effect on cytokine gene expression of all the sample groups were increased than control group. 14. In flow cytometry there's no significant relation in number of $CD8^+$, $CD19^+$ cell, however, the number of $CD4^+$ cell and NK cell in sample groups were increased than in control group. Above the results showed that aqua-acupuncture of Cistanches Herba infusion solution has effects of anti-cancer, anti-metastasis and immune response improvement.

  • PDF

천식 마우스 모델의 알러지 반응에서 Enterococcus faecalis 전체 추출물의 억제 효과 (The Whole Extract of Enterococcus faecalis Has Suppressive Effect on the Allergic Responses in Asthmatic Mouse Model)

  • 장정현;양은주;유선녕;안순철
    • 생명과학회지
    • /
    • 제27권10호
    • /
    • pp.1168-1175
    • /
    • 2017
  • 프로바이오틱스(probiotics)는 숙주에 이로운 작용을 하는 장내세균으로 주로 알려져 있으며 꾸준히 염증치료의 새로운 치료물질로 주목 받고 있다. 그러나 Lactobacillus나 Bifidobacterium과 같은 전형적인 프로바이오틱스는 장내에서 꾸준히 생존하기 어렵고 그로 인해 매개할 수 있는 다양한 효능이 줄어든다. 반면, Enterococcus는 다양한 물질을 분비하고 생물학적 활성을 지니고 있는 프로바이오틱스이긴 하나 동시에 기회감염균으로 항생제 내성능을 쉽게 획득할 수도 있어 사용을 자제할 것을 권고하고 있으며, Vancomycin 내성인자를 원래부터 가지고 있어서 이 인자의 발현과 함께 타 독성균주에 전달할 가능성이 있다. 그러나 본 연구에서는 이러한 Enterococcus의 추출물을 이용하여 항염증 물질을 확인해보고자 하였다. 특히, 호흡기계 질환인 천식의 치료효능을 보기 위해 human monocytic cell line인 THP-1 세포를 이용한 in vitro test와 천식 유발 BALB/c mice를 이용한 in vivo test를 시행하였다. 천식은 사이토카인과 IgE 등 세포 염증성 물질을 분비하고 호산구, 호중구, 호염기구, 비만세포, 단구, T 세포 등 다양한 염증 세포가 폐 부위로 침윤되는 복합적인 염증 질환으로 잘 알려져 있다. 본 연구에서는 우선 Enterococcus faecalis 에서 초음파 처리를 통해 균체성분을 분해한 다음 동결건조하여 추출물을 획득하였다. E. faecalis에서 추출한 추출물(Ef extract)을 PBS에 부유시킨 다음, 염증유발을 위해 ovalbumin (OVA)를 처리한 THP-1 세포에 처리한 결과, OVA에 의해 증가한 THP-1 세포의 생존율이 감소하였다. 또한 OVA에 의해 천식이 유발된 BALB/c mice에서 염증부위인 폐의 세척액을 채취하여 폐 조직 내 세포 변화를 관찰하였다. 그 결과, Ef extract에 의해 호산구와 호중구가 Ef extract 농도에 따라 점차 감소하였고 혈액 내에서도 Ef extract에 의해 호산구, 호중구, 호염기구가 감소하였다. 이는 Ef extract에 의해 염증세포로부터 IL-5의 분비를 억제하고 IgE의 양을 감소시켜 이와 관련된 염증세포의 침윤 및 증가를 억제한 것으로 사료된다. 이를 통해 Ef extract 가 천식 동물모델에서 항염증효과가 있음을 관찰하였지만 Ef extract의 구성 물질 분석 및 작용 메커니즘에 대한 추가연구를 통해 천식 치료물질로의 가능성을 규명할 것이다.