基于斑马鱼模型的知母皂苷BⅡ的血管保护作用及机制研究
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篇名: 基于斑马鱼模型的知母皂苷BⅡ的血管保护作用及机制研究
TITLE: Study on Vascular Protection Effect and Mechanism of Timosaponin B Ⅱ Based on Zebrafish Model
摘要: 目的:研究知母皂苷BⅡ(TB-Ⅱ)的血管保护作用,并探讨其可能的作用机制。方法:以养殖水为空白对照,考察100、200和400μg/mLTB-Ⅱ培养受精后24h(24hpf)的正常斑马鱼胚胎48h后对其肠下静脉血管(SIVs)的影响。采用酪氨酸激酶抑制剂PTK787(0.06μg/mL)诱导斑马鱼肠下血管损伤模型;以0.1%二甲基亚砜、不加PTK787为空白对照,加PTK787、不加药物为模型对照,考察100、200、400μg/mLTB-Ⅱ作用48h后对血管损伤模型斑马鱼SIVs的影响,并采用实时荧光定量-聚合酶链式反应法检测斑马鱼体内fam样酪氨酸激酶1(Flt-l)、含激酶插入区受体(Kdr)、含激酶插入区受体(lKdr-l)、血管内皮生长因子A(VEGF-A)以及肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)mRNA的相对表达量。结果:100μg/mLTB-Ⅱ作用后可显著增加正常斑马鱼的SIVs出芽数(P<0.05),200μg/mLTB-Ⅱ作用后可显著增加正常斑马鱼的SIVs数(P<0.05)。与空白对照比较,PTK787作用后斑马鱼SIVs数显著减少(P<0.01),Flt-l、Kdr、Kdr-l、VEGF-A、TNF-α和IL-6mRNA的相对表达量均显著降低(P<0.05或P<0.01);而经不同质量浓度的TB-Ⅱ作用后,血管损伤模型斑马鱼的SIVs数均不同程度地增加,Flt-l、Kdr、Kdr-l、VEGF-A、TNF-α和IL-6mRNA相对表达量均不同程度地升高,除100μg/mLTB-Ⅱ作用后斑马鱼SIVs数和Flt-l、TNF-αmRNA表达量以及400μg/mLTB-Ⅱ作用后斑马鱼TNF-αmRNA表达量增加不显著外,其余指标差异均有统计学意义(P<0.05或P<0.01)。结论:TB-Ⅱ具有一定的促血管新生和修复受损血管的作用,其机制可能与上调血管内皮生长因子受体和促炎细胞因子的表达相关。
ABSTRACT: OBJECTIVE:To study the protective effect of timosaponin BⅡ(TB-Ⅱ)on blood vessels and explore its possible mechanism. METHODS :Using aquaculture water as blank control ,the effects of 100,200 and 400 μg/mL TB-Ⅱ treatment for 48 h on the situation of subintestinal veins (SIVs)in normal zebrafish embryos 24 h after fertilization (24 hpf)were investigated. PTK787(0.06 μg/mL),a tyrosine kinase inhibitor ,was used to induce the model of zebrafish intestinal vascular injury ;using combing with 0.1% dimethyl sulfoxide but no PTK 787 as blank control ,combing with PTK 787 but no drug as model control ,the effects treatment of 100,200 and 400 μg/mL TB-Ⅱ for 48 h on the SIVs of zebrafish model with vascular injury were investigated. Relative expressions of fam-like tyrosine kinase 1(Flt-1),kinase insert domain containing receptor (Kdr),kinase insert domain containing receptor l (Kdr-l),vascular endothelial growth factor A (VEGF-A),tumor necrosis factor α(TNF-α)and interleukin 6 (IL-6)mRNA were detected by RT-PCR. RESULTS :100 μg/mL TB-Ⅱ could significantly increase the sprouting vessel of normal zebrafish SIVs sprouting vessel number (P<0.05),and 200 μg/mL TB-Ⅱ could significantly increase SIVs number of normal zebrafish (P<0.05). Compared with blank control , SIVs treatment (P<0.01),and the relative expressions of Flt-l , Kdr,Kdr-l,VEGF-A,TNF-α and IL-6 mRNA were alse decreased significantly (P<0.05 or P<0.01). After treated 化。E-mail:pn333@163.com with different concentrations of TB- Ⅱ ,SIVs number of vascular injury model zebrafish increase d to different extents ;relative expressions of Flt-l ,Kdr,Kdr-l,VEGF-A,TNF-α and IL-6 mRNA were increased to different extents. There was no significant difference in SIVs number and the expression of Flt-l ,TNF-α mRNA in zebrafish treated with 100 μg/mL TB-Ⅱ and the expression of TNF-α mRNA in zebrafish treated with 400 μg/mL TB-Ⅱ, but there was statistical significance in other indexes (P<0.05 or P<0.01). CONCLUSIONS :TB-Ⅱ has a certain function of promoting angiogenesis and repairing damaged blood vessels ,and its mechanism is related to the up-regulation of vascular endothelial growth factor receptor and pro-inflammatory cytokine expression.
期刊: 2020年第31卷第07期
作者: 杜孟姣,陈坚平,余嘉贤,梅文杰,余楚钦,王延东
AUTHORS: DU Mengjiao ,CHEN Jianping ,YU Jiaxian ,MEI Wenjie ,YU Chuqin ,WANG Yandong
关键字: 知母皂苷BⅡ;血管保护;斑马鱼;肠下静脉血管;机制
KEYWORDS: Timosaponin B Ⅱ;Vascular protection ;Zebrafish;Subintestinal veins ;Mechanism
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