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  

Study on Vascular Protection Effect and Mechanism of Timosaponin BⅡBased on Zebrafish Model

作　　者：杜孟姣 陈坚平[2] 余嘉贤 梅文杰[1] 余楚钦[1] 王延东[4] DU Mengjiao;CHEN Jianping;YU Jiaxian;MEI Wenjie;YU Chuqin;WANG Yandong(Guangdong Pharmaceutical University/Guangdong Provincial Key Laboratory of Advanced Drug Delivery Systems/Guangdong Provincial Molecular Probe and Biomedical Imaging Engineering Center/Guangdong Provincial Engineering Center of Topical Precise Drug Delivery System,Guangzhou 510006,China;Dept.of Pharmacy,the First Hospital Affiliated to Sun Yat-sen University,Guangzhou 510080,China;School of Stomatology,Jinan University,Guangzhou 510632,China;Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou 510060,China)

机构地区：[1]广东药科大学/广东省药物新剂型重点实验室/广东省分子探针与生物医学影像工程中心/广东省局部精准药物递药制剂工程技术研究中心,广州510006 [2]中山大学附属第一医院药学部,广州510080 [3]暨南大学口腔医学院,广州510632 [4]中山大学中山眼科中心,广州510060

出　　处：《中国药房》2020年第7期811-815,共5页China Pharmacy

基　　金：广东省科学技术厅-广东省中医药科学院联合科研项目(No.2016A020226038)。

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/mL TB-Ⅱ培养受精后24 h(24 hpf)的正常斑马鱼胚胎48 h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/mL TB-Ⅱ作用48 h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l(Kdr-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/mL TB-Ⅱ作用后可显著增加正常斑马鱼的SIVs出芽数(P<0.05),200μg/mL TB-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-6 mRNA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-6 mRNA相对表达量均不同程度地升高,除100μg/mL TB-Ⅱ作用后斑马鱼SIVs数和Flt-l、TNF-αmRNA表达量以及400μg/mL TB-Ⅱ作用后斑马鱼TNF-αm RNA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。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 400μg/mL TB-Ⅱtreatment for 48 h on the situation of subintestinal veins(SIVs)in normal zebrafish embryos 24 h after fertilization(24 hpf)were investigated.PTK787(0.06μg/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 PTK787 as blank control,combing with PTK787 but no drug as model control,the effects treatment of 100,200 and 400μg/mL TB-Ⅱfor 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 factorα(TNF-α)and interleukin 6(IL-6)mRNA were detected by RT-PCR.RESULTS:100μg/mL TB-Ⅱcould significantly increase the sprouting vessel of normal zebrafish SIVs sprouting vessel number(P<0.05),and 200μg/mL TB-Ⅱcould significantly increase SIVs number of normal zebrafish(P<0.05).Compared with blank control,SIVs number of zebrafish decreased significantly after PTK787 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 with different concentrations of TB-Ⅱ,SIVs number of vascular injury model zebrafish increased to different extents;relative expressions of Flt-l,Kdr,Kdr-l,VEGF-A,TNF-αand IL-6 mRNA were increased to different extents.There was no significant difference in SIVs number and the expression of Flt-l,TNF-αmRNA in zebrafish treated with 100μg/mL TB-Ⅱand the expression of TNF-αmRNA in zebrafish treated with 400μg/mL TB-Ⅱ,but there was statistical significance in other indexes(P<0.05 or P<0.01).CONCLUSIONS:TB-Ⅱhas a certain function of promoting angiogenesis an

关 键 词：知母皂苷BⅡ 血管保护 斑马鱼 肠下静脉血管 机制 

分 类 号：R285.5[医药卫生—中药学]