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目的:探讨Jagged1/Notch信号通路在血府逐瘀胶囊调控血管新生中的作用。方法:以2.5%的血府逐瘀胶囊含药血清和空白血清分别干预人微血管内皮细胞(HMEC-1),通过实时荧光定量PCR(Real-time PCR)检测药物处理细胞12,24,36,48 h时Jagged1 mRNA水平的表达,通过蛋白免疫印迹法(Western blot)检测药物处理细胞12,24,48 h时Jagged1在蛋白质水平的表达;在血府逐瘀胶囊促血管新生最佳药效条件下,使用γ-分泌酶抑制剂(3,5-二氟苯乙酰基)-L-丙氨酰基-L-2-苯基甘氨酸叔丁酯(DAPT)阻断Notch信号通路,设置DAPT处理组,二甲基亚砜(DMSO)溶剂组和空白组,通过体外成血管实验检测抑制Jagged1/Notch信号对药物诱导HMEC-1体外成血管能力的影响。结果:血府逐瘀胶囊含药血清干预内皮细胞12 h能上调Jagged1在mRNA水平表达(P<0.01),药物干预细胞24 h能同时上调Jagged1在mRNA和蛋白水平表达(P<0.05);DAPT阻断Notch信号后,药物促体外成血管能力下降。结论:血府逐瘀胶囊可通过调控Jagged1表达,影响Jagged1/Notch信号通路,从而调控血管新生。
Objective: To investigate the role of Jagged1 / Notch signaling pathway in the regulation of angiogenesis by Xuefu Zhuyu Capsule. Methods: Human microvascular endothelial cells (HMEC-1) were treated with 2.5% serum containing Xuefu Zhuyu Capsule and blank serum respectively. The cells treated with drugs were treated with Real-time PCR (Real-time PCR) 12,24,36, At 48 h, the expression of Jagged1 mRNA was detected by Western blot. The protein expression of Jagged1 was detected at 12, 24 and 48 h after drug treatment. The optimal pharmacodynamic parameters of Xuefuzhuyu capsule in promoting angiogenesis The Notch signaling pathway was blocked using the γ-secretase inhibitor (3,5-difluorophenylacetyl) -L-alanyl-L-2-phenylglycine tert-butyl ester (DAPT) , Dimethylsulfoxide (DMSO) solvent group and blank group. The effect of inhibiting Jagged1 / Notch signal on the in vitro angiogenesis of HMEC-1 was tested by in vitro vascularization assay. Results: The intervention of Xuefuzhuyu capsule with serum for 12 h could up-regulate the expression of Jagged1 at the mRNA level (P <0.01), and the mRNA expression of Jagged1 at the mRNA and protein level increased 24 h after treatment (P <0.05); DAPT After blocking the Notch signal, the drug pro-angiogenic vascular capacity decreased. Conclusion: Xuefu Zhuyu Capsule can regulate angiogenesis by regulating Jagged1 expression and affecting Jagged1 / Notch signaling pathway.