• 제목/요약/키워드: Animal disease model

검색결과 443건 처리시간 0.026초

Generation of ints14 Knockout Zebrafish using CRISPR/Cas9 for the Study of Development and Disease Mechanisms

  • Ji Hye Jung;Sanghoon Jeon;Heabin Kim;Seung-Hyun Jung
    • 한국발생생물학회지:발생과생식
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    • 제27권4호
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    • pp.205-211
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    • 2023
  • INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.

도라지 추출물 연양갱이 β-amyloid에 의한 세포독성 및 Scopolamine에 의해 유도된 인지능 저하 동물 모델의 개선효과 (Improving Effects of Platycodon Extracts Jelly on β-amyloid-induced Cytotoxicity and Scopolamine-Induced Cognitive Impairment Animal Models)

  • 오홍근;김정훈;신은혜;강영례;이봉근;박상훈;문대인;권이성;김영필;최민휴;김옥진;박광현;이학용
    • 한국자원식물학회지
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    • 제26권4호
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    • pp.417-425
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    • 2013
  • 본 연구에서는 도라지 추출물에 대한 세포 독성을 확인하였으며, $A{\beta}$에 의한 PC12 세포 독성에 대한 보호효과를 관찰하였다. 또한 도라지 추출물과 도라지 추출물 연양갱을 4주간 강제 경구 투여하여 Morris 수중미로시험에서 도달지점까지의 도달시간이 도라지 추출물 및 도라지 추출물 연양갱 투여에 의해서 모두에서 유의하게 감소하였다. 이와 유사하게 수동회피시험에서도 자극이 있는 어두운 방을 나오는 시간이 도라지 추출물 및 도라지 추출물 연양갱 투여에 의해서 현저하게 감소하였다. 따라서 도라지 추출물 및 도라지 추출물 연양갱은 인지능 개선에 도움을 줄 것으로 생각된다.

Preparation of Alzheimers Animal Model and Brain Dysfunction Induced by Continuous $\beta$-Amyloid Protein Infusion

  • Akio Itoh;Kiyofumi Yamada;Kim, Hyoung-Chun;Toshitaka Nabeshima
    • Toxicological Research
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    • 제17권
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    • pp.47-57
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    • 2001
  • Alzheimer's disease (AD) is the most common cause of dementia in the elderly, and its pathology is characterized by the presence of numerous numbers of senile plaques and neurofibrillary tangles. Several genetic and transgenic studies have indicated that excess amount of $\beta$-amyloid protein (A$\beta$) is produced by mutations of $\beta$TEX>$\beta$-amyloid precursor protein and causes learning impairment. Moreover, $A\beta$ has a toxic effect on cultured nerve cells. To prepare AD model animals, we have examined continuous (2 weeks) infusion of $A\beta$ into the cerebral ventricle of rats. Continuous infusion of $A\beta$ induces learning impairment in water maze and passive avoidance tasks, and decreases choline acetyltransferase activity in the frontal cortex and hippocampus. Immunohistochemical analysis revealed diffuse depositions of $A\beta$ in the cerebral cortex and hippocampus around the ventricle. Furthermore, the nicotine-evoked release of acetylcholine and dopamine in the frontal cortex/hippocampus and striatum, respectively, is decreased in the $A\beta$-infused group. Perfusion of nicotine (50 $\mu\textrm{M}$) reduced the amplitude of electrically evoked population spikes in the CA1 pyramidal cells of the control group, but not in those of the $A\beta$-infused group, suggesting the impairment of nicotinic signaling in the $A\beta$-infused group. In fact, Kd, but not Bmax, values for [$^3H$] cytisine binding in the hippocampus significantly increased in the $A\beta$-infused rats. suggesting the decrease in affinity of nicotinic acetylcholine receptors. Long-term potentiation (LTP) induced by tetanic stimulations in CA1 pyramidal cells, which is thought to be an essential mechanism underlying learning and memory, was readily observed in the control group, whereas it was impaired in the $A\beta$-infused group. Taken together, these results suggest that $A\beta$ infusion impairs the signal transduction mechanisms via nicotinic acetylcholine receptors. This dysfunction may be responsible, at least in part, for the impairment of LTP induction and may lead to learning and memory impairment. We also found the reduction of glutathione- and Mn-superoxide dismutase-like immunoreactivity in the brains of $A\beta$-infused rats. Administration of antioxidants or nootropics alleviated learning and memory impairment induced by $A\beta$ infusion. We believe that investigation of currently available transgenic and non-transgenic animal models for AD will help to clarify the pathogenic mechanisms and allow assessment of new therapeutic strategies.

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RAW 264.7 대식세포와 염증유도 동물모델에서 산겨릅나무 추출물의 항산화 및 항염증 효과 (In Vitro and In Vivo Anti-Oxidative and Anti-Inflammatory Activities of Acer tegmentosum Maxim Extracts)

  • 이초은;정현희;조진아;이선영
    • 한국식품영양과학회지
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    • 제46권1호
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    • pp.1-9
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    • 2017
  • 산겨릅나무 $80^{\circ}C$ 열수 및 95% 에탄올 추출물을 제조하여 항산화 및 항염증 효과를 확인한 결과 추출물은 우수한 항산화 효과를 나타내었고 RAW 264.7 세포에서도 항염증 효과를 보였으며, 그중 95% 에탄올 추출물의 효능이 더 큰 것으로 나타났다. 따라서 효능이 더 뛰어난 산겨릅나무 에탄올 추출물을 이용하여 in vivo 실험에서 추출물의 세포 보호효과 및 항염증 효능을 재확인하였다. ICR mouse에 일주일간 100 mg/kg B.W 산겨릅나무 에탄올 추출물을 경구투여한 후 LPS를 처리하여 염증반응 및 산화적 스트레스를 유도시켜 Comet assay, serum tumor necrosis $factor(TNF)-{\alpha}$$interferon-{\gamma}$, 장점막세포에서의 $TNF-{\alpha}$와 interleukin-6에 대한 생성능을 측정한 결과 산겨릅나무 에탄올 추출물은 DNA 손상을 억제하였으며 사이토카인 생성을 유의적으로 억제하여 항산화와 항염증 효과를 보였다. 따라서 본 연구에서 사용한 산겨릅나무의 에탄올 추출물은 산화적 스트레스 및 염증을 억제하는 효과를 나타내는 세포보호 기능적 소재로서 가능성이 있는 것으로 생각한다.

인공심장판막의 개발과 동물실험 -인공심장판막의 2차 동물실험- (The Second Animal Tests of Artificial Heart Valves)

  • 김형묵
    • Journal of Chest Surgery
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    • 제23권4호
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    • pp.617-621
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    • 1990
  • A heart supplies blood of about 15, 000 liters to each human organ in a day. A normal function of heart valves is necessary to accomplish these enormous work of heart. The disease of heart valve develops to a narrowness of a closure, resulting in an abnormal circulation of blood. In an attempt to eliminate the affliction of heart valves, the operative method to replace with artificial heart valves has developed and saved numerous patients over past 30 years. This replacement operation has been performed since early 1960`s in Korea, but all the artificial heart valves used are imported from abroad with very high costs until recent years. New artificial heart valves have been developed in Korea Advanced Institute of Science and Technology since early 1980`s. The first developed valve was designed with a free-floating pyrolytic carbon disk that is suspended in a titanium cage. The design of the valve was tested in vitro, and in animals in 1987. The results from this study was that the eccentrically placed struts creates a major and minor orifice when the disc opens and stagnation of flow in the area of the minor orifice has led to valve thrombosis. In this work, the design of the valve was changed from a single - leaflet valve to double - leaflet one in order to resolve the problems observed in the first - year tests. Morphological and hemodynamic studies were made for the newly designed valves through the in vitro and in vivo tests. The design and partial materials of the artificial heart valve was improved comparing with first - year`s model. The disc in the valve was modified from single - leaflet to bi - leaflet, and the material of the cage was changed from titanium metal to silicon - alloyed pyrolytic carbon. A test was made for the valve in order to examine its mechanical performance and stability. Morphological and hemodynamic studies were made for the valve that had been implanted in tricuspid position of mongrel dogs. All the test animals were observed just before the deaths. A new artificial heart valve was designed and fabricated in order to resolve the problems observed in the old model. The new valve was verified to have good stability and high resistance to wear through the performance tests. The hemodynamic properties of the valve after implantation were also estimated to be good in animal tests. Therefore, the results suggest that the newly designed valve in this work has a good quality in view of the biocompatibility. However, valve thrombosis on valve leaflets and annulus were found. This morphological findings were in accordance with results of surface polishing status studies, indicating that a technique of fine polishing of the surface is necessary to develop a valve with higher quality and performance.

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Sustained release of alginate hydrogel containing antimicrobial peptide Chol-37(F34-R) in vitro and its effect on wound healing in murine model of Pseudomonas aeruginosa infection

  • Shuaibing Shi;Hefan Dong;Xiaoyou Chen;Siqi Xu;Yue Song;Meiting Li;Zhiling Yan ;Xiaoli Wang ;Mingfu Niu ;Min Zhang;Chengshui Liao
    • Journal of Veterinary Science
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    • 제24권3호
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    • pp.44.1-44.17
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    • 2023
  • Background: Antibiotic resistance is a significant public health concern around the globe. Antimicrobial peptides exhibit broad-spectrum and efficient antibacterial activity with an added advantage of low drug resistance. The higher water content and 3D network structure of the hydrogels are beneficial for maintaining antimicrobial peptide activity and help to prevent degradation. The antimicrobial peptide released from hydrogels also hasten the local wound healing by promoting epithelial tissue regeneration and granulation tissue formation. Objective: This study aimed at developing sodium alginate based hydrogel loaded with a novel antimicrobial peptide Chol-37(F34-R) and to investigate the characteristics in vitro and in vivo as an alternative antibacterial wound dressing to treat infectious wounds. Methods: Hydrogels were developed and optimized by varying the concentrations of crosslinkers and subjected to various characterization tests like cross-sectional morphology, swelling index, percent water contents, water retention ratio, drug release and antibacterial activity in vitro, and Pseudomonas aeruginosa infected wound mice model in vivo. Results: The results indicated that the hydrogel C proved superior in terms of cross-sectional morphology having uniformly sized interconnected pores, a good swelling index, with the capacity to retain a higher quantity of water. Furthermore, the optimized hydrogel has been found to exert a significant antimicrobial activity against bacteria and was also found to prevent bacterial infiltration into the wound site due to forming an impermeable barrier between the wound bed and external environment. The optimized hydrogel was found to significantly hasten skin regeneration in animal models when compared to other treatments in addition to strong inhibitory effect on the release of pro-inflammatory cytokines (interleukin-1β and tumor necrosis factor-α). Conclusions: Our results suggest that sodium alginate -based hydrogels loaded with Chol-37(F34-R) hold the potential to be used as an alternative to conventional antibiotics in treating infectious skin wounds.

개에서 건삭파열로 유발한 급성 이첨판 폐쇄부전 모델의 혈류역학적 평가 (Hemodynamic Evaluation of Acute Mitral Valve Insufficiency Model induced by Chordae Tendinae Rupture in Normal Dogs)

  • 김세훈;김남수;이기창;김종민;김민수
    • 한국임상수의학회지
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    • 제31권5호
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    • pp.367-370
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    • 2014
  • 본 연구는 개에서 건삭파열로 유발한 급성 이첨판 폐쇄부전 모델의 혈류역학적 변화를 관찰하는 것이다. 이 연구는 10마리의 정상 심장기능을 가진 비글견에서 실시되었다. 직접혈압측정법과 Swan-Ganz 카테터를 통해 건삭을 실험적으로 파열시키는 기간동안 혈류역학적 지표 변화를 진행하였다. 이첨판 폐쇄부전 모델을 만들기 위해 5번 늑간으로 접근하여 관절경에 사용하는 작은 훅나이프를 사용하여 건삭을 파열하였다. 수술 중 칼라 도플러 영상 검사를 통해 이첨판 역류를 확인하였다. 혈류역학적 지표를 측정한 결과 폐모세혈관쇄기압은 유의적으로 증가하였지만, 평균동맥압, 정맥압, 폐동맥압, 심박출량, 심박출 지수는 건삭 파열 후에 유의적으로 감소한다는 것을 발견하였다. 이것은 건산파열로 인해 좌심실로 부터 역류된 혈액이 좌심방에 과부하를 일으킨다는 것을 나타내는 것이다. 본 연구를 통해 개에서 급성 이첨판 폐쇄부전 모델을 만드는데 있어 건삭파열방법이 효과적인 것을 알 수 있었고, 이첨판 역류 유발 후 시간이 지나면 만성 이첨판 부전증으로 진행할 수 있을 것으로 생각되며, 차후 판막의 만성적 변화 양상을 연구하는데 도움을 줄 수 있을 것으로 생각됩니다.

DSS로 유도된 염증성 장 질환 동물 모델에서 황금 열수 추출물이 면역 조절 기능에 미치는 영향 (Immunoregulatory Effects of Water Extracts of Scutellariae Radix in DSS-Induced Inflammatory Bowel Disease Animal Model)

  • 이순희;임병우;조여원
    • Journal of Nutrition and Health
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    • 제37권6호
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    • pp.431-439
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    • 2004
  • Scutellariae Radix (Scu.), one of the immune-regulatory substances, is recognized to play the role in the metabolic process of inflammation, allergy and immunity. It has been traditionally used in the Oriental medicine to treat inflammatory bowel diseases (IBD). The purpose of this study was to evaluate the effects of water extracts of Scutellariae Radix on the spleen lymphocyte immune function in the Balb/c female mice treated with dextran sodium sulfate (DSS) to induce colitis. Water extract of Scutellariae Radix (100 mg/kg) and sulfasalazine (50 mg/kg) were administrated orally for 2 weeks of experimental period. Mice were divided into three experimental groups randomly: DSS group (5% DSS was ad libitum for 5 days) as control group, DSS + Scu. (water extracts of Scutellariae Radix for 2 weeks after 5% DSS was ad libitum for 5 days) as experimental group, and DSS + Sulfasalazine group (Sulfasalazine for 2 weeks after 5% DSS was ad libitum for 5 days) as positive control group. Levels of Ig A, Ig E, CD4$^{+}$, CD8$^{+}$, TNF-$\alpha$ and other cytokines were measured. Treatment of DSS for 5 days induced bowel inflammation and the treatment with Scu. water exteract and sulfasalazine significantly recovered the damage. The length of intestine of DSS group was significantly shorter than that of other groups. The serum and fecal concentration of Ig A of SS + Scu group was higher than those of DSS group. The contents of CD4$^{+}$ T cells was higher in the DSS + Scu. group than the other groups and CD8$^{+}$ T cells was the lowest in DSS + Sulfasalazine group. The Ig A level of cultured supernatant of spleen lymphocyte was the highest, while the Ig E level was the lowest in SS + Scu group. The concentration of TNF-$\alpha$, cytokine secreted from the Th1 cell in the supernatant spleen lymphocyte, was the highest in the DSS group and the lowest in the DSS + Scu. group. The concentration of IFN-${\gamma}$ and ll...-12 was lower in the DSS + Scu. group than those of the other groups. The concentration of IL-4 in the supernatant of spleen lymphocyte was the lowest in the DSS + Scu. group but IL-10 was not significantly different. Based on these findings, water extract of Scutellariae Radix exhibited the inhibitory effect via IL-4 production thereby inhibited the production of Ig E and strengthened immune system, and alleviated injury in DSS- induced colitis mice model.

Restorative effects of Rg3-enriched Korean Red Ginseng and Persicaria tinctoria extract on oxazolone-induced ulcerative colitis in mice

  • Ullah, H.M. Arif;Saba, Evelyn;Lee, Yuan Yee;Hong, Seung-Bok;Hyun, Sun-Hee;Kwak, Yi-Seong;Park, Chae-Kyu;Kim, Sung Dae;Rhee, Man Hee
    • Journal of Ginseng Research
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    • 제46권5호
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    • pp.628-635
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    • 2022
  • Background: Ulcerative colitis (UC) is the large intestine disease that results in chronic inflammation and ulcers in the colon. Rg3-enriched Korean Red Ginseng extract (Rg3-RGE) is known for its pharmacological activities. Persicaria tinctoria (PT) is also used in the treatment of various inflammatory diseases. The aim of this study is to investigate the attenuating effects of Rg3-RGE with PT on oxazolone (OXA)-induced UC in mice. Methods: A total of six groups of mice including control group, OXA (as model group, 1.5%) group, sulfasalazine (75 mg/kg) group, Rg3-RGE (20 mg/kg) group, PT (300 mg/kg) group, and Rg3-RGE (10 mg/kg) with PT (150 mg/kg) group. Data on the colon length, body weight, disease activity index (DAI), histological changes, nitric oxide (NO) assay, Real-time PCR of inflammatory factors, ELISA of inflammatory factors, Western blot, and flow cytometry analysis were obtained. Results: Overall, the combination treatment of Rg3-RGE and PT significantly improved the colon length and body weight and decreased the DAI in mice compared with the treatment with OXA. Additionally, the histological injury was also reduced by the combination treatment. Moreover, the NO production level and inflammatory mediators and cytokines were significantly downregulated in the Rg3-RGE with the PT group compared with the model group. Also, NLR family pyrin domain containing 3 (NLRP3) inflammasome and nuclear factor kappa B (NF-𝛋B) were suppressed in the combination treatment group compared with the OXA group. Furthermore, the number of immune cell subtypes of CD4+ T-helper cells, CD19+ B-cells, and CD4+ and CD25+ regulatory T-cells (Tregs) was improved in the Rg3-RGE with the PT group compared with the OXA group. Conclusion: Overall, the mixture of Rg3-RGE and PT is an effective therapeutic treatment for UC.

트립톨라이드가 식이제한에 의한 수명연장과 노화관련 질환에 미치는 영향 (Triptolide Mimics the Effect of Dietary Restriction on Lifespan and Retards Age-related Diseases in Caenorhabditis elegans)

  • 백선미;박상규
    • 생명과학회지
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    • 제28권8호
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    • pp.931-937
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    • 2018
  • 뇌공등에 함유되어있는 트립톨라이드는 뛰어난 항염증, 항산화 효능을 가지고 있음이 보고되었다. 예쁜꼬마선충을 이용한 이전 연구에서 트립톨라이드의 섭취가 개체의 항스트레스 효능을 높이고, 수명을 연장시킴이 밝혀졌다. 본 연구에서는 트립톨라이드에 의한 수명연장에 관여하는 세포 내 기전과 트립톨라이드가 노화관련 질환인 당뇨병과 알츠하이머병에 미치는 영향을 평가하였다. 트립톨라이드는 인슐린/IGF-1-like 신호전달 저하에 의한 수명연장 돌연변이인 age-1과 미토콘드리아 전자 전달계 저하에 의한 수명연장 돌연변이인 clk-1의 수명을 유의적으로 증가시킨 반면, 식이제한 유도 돌연변이인 eat-2의 수명에는 유의적인 변화를 유도하지 못했다. 또한 박테리아 희석을 이용한 식이제한에 의해 연장된 수명을 추가적으로 더 연장시키지 못했다. 트립톨라이드 섭취는 고농도의 당 섭취에 의한 체내 독성과 사람 아밀로이드 베타 형질전환 유전자로 인한 체내 독성을 유의적으로 저하시켰다. 이러한 결과들은 트립톨라이드에 의한 수명연장이 식이제한에 의한 수명연장 기전과 중복되며, 트립톨라이드가 노화관련 질환을 저해하는 효능이 있음을 보여준다. 따라서, 트립톨라이드는 식이제한 효능을 대체할 수 있는 식의약품 개발에 활용될 수 있다.