ex vivo

离体
  • 文章类型: Journal Article
    在这项研究中,使用了20至80岁无皮肤病的韩国成年人的头皮组织。处理头皮组织,用不同的处理方法(包括Bioscalp,UltraExo助推器,和UltraSLinePlus)来自UltraV公司。超过12天,每隔一段时间(第0、3、6、9和12天)记录毛囊特征的观察和测量结果。该研究评估了这些物质对毛囊生长和形态的影响。Bioscalp,结合UltraExoBooster和UltraSLinePlus,在离体显示显著的毛发伸长。观察到毛球直径的保存,表明基于外泌体的产品具有持续毛发生长的潜力。毛发生长周期分析表明,与未处理组相比,在来自UltraV的测试产品中更低的向退步阶段的过渡。研究结果表明,所测试的配方,尤其是Bioscalp的组合,UltraExo助推器,和UltraSLinePlus,在促进头发生长方面表现出显著的效果,保持毛球的完整性,并减少向退步阶段的过渡。这项研究提出了有希望的脱发替代疗法,说明结果与常规测试产品组一样好。
    In this study, scalp tissues from Korean adults between 20 and 80 without skin disease were used. Scalp tissues were processed, and hair follicles were isolated and cultured with different treatments (including Bioscalp, Ultra Exo Booster, and Ultra S Line Plus) from Ultra V company. Over 12 days, observations and measurements of hair follicle characteristics were recorded at intervals (Days 0, 3, 6, 9, and 12). The study assessed the impact of these substances on hair follicle growth and morphology. Bioscalp, combined with Ultra Exo Booster and Ultra S Line Plus, showed significant hair elongation in ex vivo. Preservation of hair bulb diameter was observed, indicating potential for sustained hair growth by exosome-based products. The hair growth cycle analysis suggested a lower transition to the catagen stage in test products from Ultra V compared to non-treated groups. The research findings indicated that the tested formulations, especially the combination of Bioscalp, Ultra Exo Booster, and Ultra S Line Plus, demonstrated significant effectiveness in promoting hair growth, maintaining the integrity of the hair bulb, and reducing the transition to the catagen stage. The study suggests promising alternative treatments for hair loss, illustrating results that were as good as those of the conventional testing product groups.
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  • 文章类型: Journal Article
    背景:下腰痛影响75%-85%的人在他们的生活中。用钠(23Na)MRI检测生化变化有可能实现更早,更准确的诊断。
    目的:为了测量离体椎间盘(IVD)中的23Na弛豫时间和表观组织钠浓度(aTSC),并研究aTSC与组织学汤普森分级之间的关系。
    方法:离体。
    方法:来自11名人体供体(4名女性,7男,平均年龄86±8岁)。
    3T;密度适应的3D径向序列(DA-3D-RAD)。
    结果:IVD23Na纵向(T1),对于每个脊柱样本(11个IVD),计算了短和长横向(T2s*和T2l*)松弛时间以及短横向松弛(ps)的比例。此外,计算所有IVD的aTSC。通过组织学Thompson分级评估IVD的降解。
    方法:在aTSC和Thompson等级之间进行KendallTau相关性(τ)检验。显著性水平设定为P<0.05。
    结果:11个IVD子集的平均23Na弛豫参数为T1=9.8±1.3毫秒,T2s*=0.7±0.1毫秒,T2l*=7.3±1.1毫秒,和ps=32.7±4.0%。总共检查了30个IVD,其中3名汤普森1级,4名2级,5名3级,5名4级,13名5级5级。aTSC随着降解的增加而降低,Thompson1级为274.6±18.9mM,Thompson5级为190.5±29.5mM。整个IVDaTSC与Thompson等级之间的相关性显着并且呈强烈负相关(τ=-0.56)。
    结论:本研究显示aTSC与退行性IVD变化之间存在显著相关性。因此,TSC有可能是有用的作为退行性脊柱变化的指标。
    方法:2技术效果:第一阶段。
    BACKGROUND: Lower back pain affects 75%-85% of people at some point in their lives. The detection of biochemical changes with sodium (23Na) MRI has potential to enable an earlier and more accurate diagnosis.
    OBJECTIVE: To measure 23Na relaxation times and apparent tissue sodium concentration (aTSC) in ex-vivo intervertebral discs (IVDs), and to investigate the relationship between aTSC and histological Thompson grade.
    METHODS: Ex-vivo.
    METHODS: Thirty IVDs from the lumbar spines of 11 human body donors (4 female, 7 male, mean age 86 ± 8 years).
    UNASSIGNED: 3 T; density-adapted 3D radial sequence (DA-3D-RAD).
    RESULTS: IVD 23Na longitudinal (T1), short and long transverse (T2s* and T2l*) relaxation times and the proportion of the short transverse relaxation (ps) were calculated for one IVD per spine sample (11 IVDs). Furthermore, aTSCs were calculated for all IVDs. The degradation of the IVDs was assessed via histological Thompson grading.
    METHODS: A Kendall Tau correlation (τ) test was performed between the aTSCs and the Thompson grades. The significance level was set to P < 0.05.
    RESULTS: Mean 23Na relaxation parameters of a subset of 11 IVDs were T1 = 9.8 ± 1.3 msec, T2s* = 0.7 ± 0.1 msec, T2l* = 7.3 ± 1.1 msec, and ps = 32.7 ± 4.0%. A total of 30 IVDs were examined, of which 3 had Thompson grade 1, 4 had grade 2, 5 had grade 3, 5 had grade 4, and 13 had grade 5. The aTSC decreased with increasing degradation, being 274.6 ± 18.9 mM for Thompson grade 1 and 190.5 ± 29.5 mM for Thompson grade 5. The correlation between whole IVD aTSC and Thompson grade was significant and strongly negative (τ = -0.56).
    CONCLUSIONS: This study showed a significant correlation between aTSC and degenerative IVD changes. Consequently, aTSC has potential to be useful as an indicator of degenerative spinal changes.
    METHODS: 2 TECHNICAL EFFICACY: Stage 1.
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  • 文章类型: Letter
    暂无摘要。
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  • 文章类型: Journal Article
    In this study we aimed to analyze the feasibility of the gasless renal biopsy technique in canine cadavers. The cadavers were randomly divided into two groups: laparoscopic GCG, in which gasless laparoscopy was performed and GCP, laparoscopy with pneumoperitoneum was performed. The procedures were randomly performed on the right and left kidneys. The total surgical time, procedural steps, and intraoperative complications were recorded. The degree of difficulty of the surgical approaches was evaluated by the surgeon, assistant, and external evaluators. Renal samples were evaluated for quality, number of glomeruli, and proportion of renal cortex. The total operative time was higher in the GCG group than in the GCP group (p < 0.01). Additionally, positioning of the second portal and platform positioning took longer than the other steps. The surgical groups differed from each other in the Likert scale values for almost all the parameters regarding the difficulty of the surgical approaches (p < 0.05), with higher scores in the GCG group than in the GCP group. Based on the video recordings, the GCP group had higher scores than the GCG group for degrees of difficulty of the approach (p < 0.05). Renal histological parameters were similar between the surgical groups and surgical sides. Our study findings indicate that the proposed gasless renal biopsy technique is feasible. The longer operative time and technical difficulties in the gasless approach did not affect the quality of the renal specimens.
    Neste estudo objetivamos analisar a viabilidade da técnica de biópsia renal gasless em cadáveres caninos. Os cadáveres foram divididos aleatoriamente em dois grupos: GCG laparoscópico, no qual foi realizada laparoscopia gasless e GCP, onde foi realizada laparoscopia com pneumoperitônio. Os procedimentos foram realizados aleatoriamente nos rins direito e esquerdo. O tempo cirúrgico total, etapas do procedimento e complicações intraoperatórias foram registrados. O grau de dificuldade das abordagens cirúrgicas foi avaliado pelo cirurgião, assistente e avaliadores externos. As amostras renais foram avaliadas quanto à qualidade, número de glomérulos e proporção do córtex renal. O tempo operatório total foi maior no grupo GCG do que no grupo GCP (p < 0,01). Além disso, o posicionamento do segundo portal e o posicionamento da plataforma demoraram mais do que as outras etapas. Os grupos cirúrgicos diferiram entre si nos valores da escala Likert para quase todos os parâmetros relativos à dificuldade das abordagens cirúrgicas (p < 0,05), com escores mais elevados no grupo GCG do que no grupo GCP. Com base nas gravações de vídeo, o grupo GCP obteve pontuações mais altas que o grupo GCG para graus de dificuldade da abordagem (p < 0,05). Os parâmetros histológicos renais foram semelhantes entre os grupos cirúrgicos e os lados cirúrgicos. Os resultados do nosso estudo indicam que a técnica proposta de biópsia renal gasless é viável. O maior tempo operatório e as dificuldades técnicas na abordagem sem gás não afetaram a qualidade das amostras renais.
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  • 文章类型: Journal Article
    关于血管老化的动物研究存在一些局限性。最重要的原因之一是缺乏快速有效的血管组织衰老模型。在这项研究中,将离体主动脉培养和Matrigel皮下植入相结合,以开发一种研究血管细胞衰老的新模型。使用8周龄的C57BL/6J小鼠获得主动脉。博来霉素用于体外诱导主动脉衰老。然后,用Matrigel将主动脉移植到受体小鼠。使用蛋白质印迹评估衰老,定量聚合酶链反应,和衰老相关的β-半乳糖苷酶活性。使用Luminex液体悬浮芯片检测炎性细胞因子。通过转录组测序分析RNA水平。结果表明,博来霉素组的血管比对照组的血管表现出明显的衰老,可以通过剥离血管外膜来增强衰老。细胞因子的水平,如白细胞介素(IL-2,IL-1β,和IL-6在离体模型中显著增加。此外,转录组测序显示,与自然老化的主动脉相比,离体模型血管中有56个显着差异表达的基因(DEG)。总之,本研究为血管细胞衰老引入了一种经济有效且节省时间的血管衰老模型。
    Animal studies on vascular aging pose a few limitations. One of the most important reasons for this is the absence of a fast and efficient model of vascular tissue aging. In this study, ex vivo aortic culture and Matrigel subcutaneous implantation are combined to develop a new model for studying vascular cellular senescence. Eight-week-old C57BL/6J mice are used to obtain aortas. Bleomycin is used to induce aortas senescence in vitro. Then, aortas are transplanted to the acceptor mice with Matrigel. Senescence is evaluated using western blotting, quantitative polymerase chain reaction, and senescence-associated beta-galactosidase activity. Inflammatory cytokines are detected using Luminex Liquid Suspension Chip. RNA levels are analyzed by transcriptome sequencing. The results revealed that vessels in the bleomycin group exhibited significant senescence than those in the control group that can be enhanced by stripping vessel adventitia. The levels of cytokines such as interleukin (IL-2, IL-1β, and IL-6 increased significantly in the ex vivo model. Furthermore, transcriptome sequencing revealed 56 significantly differentially expressed genes (DEGs) in ex vivo model vessels compared with those in naturally aging aortas. In conclusion, this study introduces a cost-effective and time-saving vessel senescence model for vascular cellular senescence.
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  • 文章类型: Journal Article
    P-糖蛋白(P-gp)是位于细胞顶端表面的重要膜转运蛋白,负责消除外源性物质和内源性物质。P-gp调节剂是可以直接或间接影响该蛋白质的化合物,导致其表达和功能的变化。这些调节剂可以作为抑制剂,诱导物,或活化剂,可能导致药物-药物相互作用(DDIs)。这篇综合综述探讨了用于评估药物诱导的P-gp调制的多种模型和技术。我们涵盖了几种方法,包括硅,在体外,离体,和体内方法,他们各自的优势和局限性。此外,我们探索涉及P-gp的DDI的治疗意义,特别关注P-gp底物的肾脏和肠道消除。这包括增强从近端肾小管上皮细胞到尿液中的有毒物质的去除或增加从肠上皮细胞到肠腔的化合物的运输。从而促进它们在粪便中的排泄。更好地理解这些相互作用,以及应用于他们研究的独特技术,对优化药物治疗至关重要,从而最大限度地减少药物引起的不良和毒性作用。
    P-glycoprotein (P-gp) is a crucial membrane transporter situated on the cell\'s apical surface, being responsible for eliminating xenobiotics and endobiotics. P-gp modulators are compounds that can directly or indirectly affect this protein, leading to changes in its expression and function. These modulators can act as inhibitors, inducers, or activators, potentially causing drug-drug interactions (DDIs). This comprehensive review explores diverse models and techniques used to assess drug-induced P-gp modulation. We cover several approaches, including in silico, in vitro, ex vivo, and in vivo methods, with their respective strengths and limitations. Additionally, we explore the therapeutic implications of DDIs involving P-gp, with a special focus on the renal and intestinal elimination of P-gp substrates. This involves enhancing the removal of toxic substances from proximal tubular epithelial cells into the urine or increasing the transport of compounds from enterocytes into the intestinal lumen, thereby facilitating their excretion in the feces. A better understanding of these interactions, and of the distinct techniques applied for their study, will be of utmost importance for optimizing drug therapy, consequently minimizing drug-induced adverse and toxic effects.
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  • 文章类型: Journal Article
    离体肩部运动模拟器通常用于研究肩部生物力学,但通常仅限于使用不允许肌肉停用的控制架构以准静态速度执行简单的平面运动。这项研究的目的是开发一种具有迭代学习和独立肌肉控制的开环肌腱偏移控制器,以功能速度模拟复杂的多平面运动,并允许肌肉失活。模拟器进行了绑架/内收,多面包绕,和外展/内收(肩胛骨下停用),使用带有植入的反向全肩关节假体的尸体肩。使用最大绝对误差(MAE)评估运动跟踪精度和可重复性,均方根误差(RMSE),和平均标准偏差(ASD)。在外展/内收和分面外展期间,海拔RMSE不超过0.3、0.7和0.8度,高程平面,和轴向旋转,分别。在绑架/内收期间,ASD不超过0.2度.外展/内收(肩胛骨下停用)导致内旋丧失,在低仰角时无法恢复。本研究提出了一种新颖的控制架构,能准确模拟复杂的肱骨关节运动。该模拟器将用作测试平台,以检查肩部病理的影响,治疗,功能性肩关节运动过程中关节生物力学的康复。
    Ex vivo shoulder motion simulators are commonly used to study shoulder biomechanics but are often limited to performing simple planar motions at quasi-static speeds using control architectures that do not allow muscles to be deactivated. The purpose of this study was to develop an open-loop tendon excursion controller with iterative learning and independent muscle control to simulate complex multiplanar motion at functional speeds and allow for muscle deactivation. The simulator performed abduction/adduction, faceted circumduction, and abduction/adduction (subscapularis deactivation) using a cadaveric shoulder with an implanted reverse total shoulder prosthesis. Kinematic tracking accuracy and repeatability were assessed using maximum absolute error (MAE), root mean square error (RMSE), and average standard deviation (ASD). During abduction/adduction and faceted circumduction, the RMSE did not exceed 0.3, 0.7, and 0.8 degrees for elevation, plane of elevation, and axial rotation, respectively. During abduction/adduction, the ASD did not exceed 0.2 degrees. Abduction/adduction (subscapularis deactivation) resulted in a loss of internal rotation, which could not be restored at low elevation angles. This study presents a novel control architecture, which can accurately simulate complex glenohumeral motion. This simulator will be used as a testing platform to examine the effect of shoulder pathology, treatment, and rehabilitation on joint biomechanics during functional shoulder movements.
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  • 文章类型: Journal Article
    背景:尽管目前有许多取石篮可用于胆管结石的取出,它们的机械性能尚未评估。本研究旨在通过检查胆管结石的机械特性来确定其特征。
    方法:这项实验研究测试了七个胆管结石回收篮的机械性能。使用专用测量装置测量径向力(RF),并且使用常规手动方法测量轴向力(AF)。
    结果:篮子之间的平均RF差异显着(p<0.001),对于VorticCatch(1.62N±0.02)和COAXIS(1.62N±0.04),其次是RASEN(1.27N±0.02),存储篮(0.95N±0.01),8线镍钛诺篮(0.93N±0.01),StoneHunter(0.78N±0.01)和花篮(0.37N±0.01),分别。篮子之间的平均AF差异显着(p<0.001),VorticCatch最高(0.668N±0.032),其次是同轴(0.629N±0.041),StoneHunter(0.574N±0.037),8线镍钛诺篮(0.546N±0.010),存储篮(0.542N±0.024),RASEN(0.435N±0.008)和花篮(0.297N±0.011),分别。根据RF和AF将篮子分为四组,其机械性能具有可比性:第1组,低RF和低AF;第2组,中度RF和中度AF;第3组,高RF和中度AF;第4组,高RF和高AF。
    结论:这项研究揭示了用于提取胆管结石的各种回收篮的不同机械性能,这可以增强对他们行动的理解。我们的结果还可以帮助将来开发检索篮。
    Although numerous retrieval baskets are currently available for the extraction of bile duct stones, their mechanical properties have not been evaluated. This study aimed to ascertain the characteristics of retrieval baskets for bile duct stones by examining their mechanical properties.
    This experimental study tested the mechanical properties of seven retrieval baskets for bile duct stones. The radial force (RF) was measured using a dedicated measurement device and the axial force (AF) was measured using the conventional manual method.
    The mean RF differed significantly among the baskets (p < 0.001) and was the strongest for VorticCatch (1.62 N ± 0.02) and COAXIS (1.62 N ± 0.04), followed by RASEN (1.27 N ± 0.02), Memory Basket (0.95 N ± 0.01), 8-wire Nitinol Basket (0.93 N ± 0.01), StoneHunter (0.78 N ± 0.01) and Flower Basket (0.37 N ± 0.01), respectively. The mean AF differed significantly among the baskets (p < 0.001) and was the highest for VorticCatch (0.668 N ± 0.032), followed by COAXIS (0.629 N ± 0.041), StoneHunter (0.574 N ± 0.037), 8-wire Nitinol Basket (0.546 N ± 0.010), Memory Basket (0.542 N ± 0.024), RASEN (0.435 N ± 0.008) and Flower Basket (0.297 N ± 0.011), respectively. The baskets were categorized into four groups with comparable mechanical properties based on the RF and AF: group 1, low RF and low AF; group 2, moderate RF and moderate AF; group 3, high RF and moderate AF; and group 4, high RF and high AF.
    This study revealed distinct mechanical properties of various retrieval baskets used for extracting bile duct stones, which may enhance the understanding of their action. Our results could also aid the development of retrieval baskets in future.
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  • 文章类型: Journal Article
    背景:囊性包虫病(CE)是全球最普遍,最重要的蠕虫人畜共患病之一。治疗主要依靠手术,或经皮干预。然而,导致复发的活的原头肌(PSC)的溢出是手术期间的问题。所以,手术前需要使用杀原剂。本研究的目的是研究微藻水醇提取物对细粒棘球蚴(s.s.)PSC的体外和离体的活性和安全性。这是对穿刺的模拟,吸气,注射,和再抽吸(PAIR)方法。
    方法:考虑到热量对桉树叶的杀原骨作用的影响,通过在80°C下的索氏提取和在室温下的渗滤进行水醇提取。通过体外和离体评估评估水醇提取物的杀原骨作用。从屠宰场收集感染的绵羊肝脏。然后,通过测序证实了包虫囊肿(HCs)的基因型,分离株仅限于S.S.在下一步中,通过扫描电子显微镜(SEM)研究了暴露于桉树的PSCs的超微结构变化。最后,通过(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)法进行细胞毒性试验,以评价E.microtheca的安全性。
    结果:通过索氏提取和渗滤制备的提取物为,在体外和离体测试中都成功地发挥了强大的杀原型作用。体外评估结果表明,通过室温渗滤(EMP)制备的小囊藻水醇提取物和通过索氏提取在80°C(EMS)制备的小囊藻水醇提取物杀死了所有PSC(100%)浓度为10和12.5mg/mL,分别。此外,与EMS相比,EMP在离体环境中20分钟后显示出99%的杀原骨作用。SEM显微照片证实了E.microtheca对PSC的有效的杀原骨和破坏作用。用MTT法测定EMP对HeLa细胞系的细胞毒性。在24小时后,50%细胞毒性浓度(CC50)的值计算为46.5μg/mL。
    结论:两种水醇提取物均显示出有效的杀原骨活性,与对照组相比,尤其是EMP产生了显着的杀原骨作用。
    BACKGROUND: Cystic echinococcosis (CE) is one of the most widespread and important global helminth zoonoses. Treatment relies mainly on surgery and, or percutaneous interventions. However, spillage of live protoscoleces (PSCs) leading to recurrence is a problem during surgery. So, the application of protoscolicidal agents before surgery is required. This study aimed to investigate the activity and safety of hydroalcoholic extracts of E. microtheca against PSCs of Echinococcus granulosus sensu stricto (s.s.) both in vitro and also ex vivo, which is a simulation to Puncture, Aspiration, Injection, and Re-aspiration (PAIR) method.
    METHODS: Considering the effects of heat on the protoscolicidal effecacy of Eucalyptus leaves, hydroalcoholic extraction was performed by both soxhlet extraction at 80 °C and percolation at room temperature. The protoscolicidal action of hydroalcoholic extracts was assessed by in vitro and ex vivo assessments. Infected sheep livers were collected from the slaughterhouse. Then, the genotype of hydatid cysts (HCs) was confirmed by sequencing and, isolates were limited to E. granulosus s.s. In the next step, ultrastructural changes of Eucalyptus-exposed PSCs were studied by scanning electron microscopy (SEM). Finally, a cytotoxicity test was conducted by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to evaluate the safety of E. microtheca.
    RESULTS: The prepared extracts by soxhlet extraction and percolation were, successfully exerted strong protoscolicidal effects in both in vitro and ex vivo tests. The results of in vitro assessment indicated that hydroalcoholic extract of E. microtheca prepared by percolation at room temperature (EMP) and hydroalcoholic extract of E. microtheca prepared by soxhlet extraction at 80 °C (EMS) killed all PSCs (100%) at concentrations of 10 and 12.5 mg/mL, respectively. Also, EMP showed 99% protoscolicidal action after 20 min in an ex vivo setting compared to EMS. SEM micrographs confirmed potent protoscolicidal and destructive effects of E. microtheca against PSCs. The cytotoxicity of EMP was tested on the HeLa cell line using MTT assay. The value of 50% cytotoxic concentration (CC50) was calculated at 46.5 μg/mL after 24h.
    CONCLUSIONS: Both hydroalcoholic extracts showed potent protoscolicidal activity and, especially EMP produced remarkable protoscolicidal effects compared to the control group.
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  • 文章类型: Journal Article
    TOTUM-070是一种专利的富含多酚的混合物,由五种不同的植物提取物组成,分别显示出对脂质代谢的潜在影响和潜在的协同作用。在这项研究中,我们调查了这种配方对健康的益处。使用高脂肪饮食的临床前模型,TOTUM-070(3g/kg体重)限制了HFD诱导的高脂血症,降低了甘油三酯(6周后-32%;12周后-20.3%)和非HDL胆固醇水平(6周后-21%;12周后-38.4%)。为了进一步研究这种益处及其在人类中的潜在机制,我们设计了一种离体临床方法来收集摄入TOTUM-070产生的循环生物活性物质,并确定它们在人肝细胞上的生物活性。在摄入TOTUM-070(4995mg)之前和之后,从健康受试者获得人血清。通过UPLC-MS/MS评估循环代谢物的存在。将含有代谢物的血清与在脂毒性环境中培养的肝细胞进一步孵育(棕榈酸酯,250µM)。RNA测序分析表明,脂质代谢是影响最大的过程之一。使用组织学,蛋白质组学,和酶分析,人TOTUM-070生物活性物质对肝细胞代谢的影响的特征是(1)抑制脂质储存,包括两种(2)甘油三酯(-41%,p<0.001)和(3)胆固醇(-50%,p<0.001)细胞内含量,(4)降低从头胆固醇合成(HMG-CoA还原酶活性-44%,p<0.001),和(5)脂肪酸合成酶蛋白水平降低(p<0.001)。总之,这些数据支持TOTUM-070对脂质代谢的有益影响,并为肝细胞中发生的人类机制提供了新的生化见解。
    TOTUM-070 is a patented polyphenol-rich blend of five different plant extracts showing separately a latent effect on lipid metabolism and potential synergistic properties. In this study, we investigated the health benefit of such a formula. Using a preclinical model of high fat diet, TOTUM-070 (3 g/kg of body weight) limited the HFD-induced hyperlipemia with a reduction in triglyceride (-32% after 6 weeks; -20.3% after 12 weeks) and non-HDL cholesterol levels (-21% after 6 weeks; -38.4% after 12 weeks). To further investigate such a benefit and its underlying mechanisms in humans, we designed an ex vivo clinical approach to collect the circulating bioactives resulting from TOTUM-070 ingestion and to determine their biological activities on human hepatocytes. Human serum was obtained from healthy subjects before and after intake of TOTUM-070 (4995 mg). The presence of circulating metabolites was assessed by UPLC-MS/MS. Serum containing metabolites was further incubated with hepatocytes cultured in a lipotoxic environment (palmitate, 250 µM). RNA sequencing analyses show that lipid metabolism was one of the most impacted processes. Using histologic, proteomic, and enzymatic assays, the effects of human TOTUM-070 bioactives on hepatocyte metabolism were characterized by (1) the inhibition of lipid storage, including both (2) triglycerides (-41%, p < 0.001) and (3) cholesterol (-50%, p < 0.001) intracellular content, (4) a reduced de novo cholesterol synthesis (HMG-CoA reductase activity -44%, p < 0.001), and (5) a lowered fatty acid synthase protein level (p < 0.001). Altogether, these data support the beneficial impact of TOTUM-070 on lipid metabolism and provide new biochemical insights in human mechanisms occurring in liver cells.
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