丹皮酚通过抑制PI3K/AKT-NF-κB通路对LPS诱导的与平滑肌细胞共培养的大鼠血管内皮细胞的保护作用

Paeonol affects proliferation activity of rat vasular endothelial cells induced by lipopolysaccharide and co-cultured with smooth muscle cells via inhibiting pathway of PI3K/AKT-NF-κB signaling

该研究为观察丹皮酚(Pae)对脂多糖(LPS)诱导的与平滑肌细胞(SMC)共培养内皮细胞(VEC)的保护作用以及对磷脂酰肌醇3激酶(phosphoinositide-3-kinase,PI3K)/丝/苏氨酸激酶(ser-ine threonine kinase,AKT)/核因子κB(nucler factorkappa B,NF-κB)信号通路的影响。采用组织块预消化贴壁法原代培养大鼠血管内皮细胞和大鼠血管平滑肌细胞;用Transwell小室建立VEC-SMC共培养模型;LPS(100μg·L^(-1),7 h)诱导VEC损伤;MTT法检测VEC活性;LDH试剂盒检测细胞通透性;Western blot法检测Pae(15,30,60μmol·L^(-1),24 h)对LPS诱导损伤的共培养VEC的PI3K/AKT及NF-κB信号通路的表达;Western blot法检测VEC的细胞核p-P65表达的影响。研究证实不同浓度的Pae可显著性抑制LPS诱导的VEC损伤,提高细胞活性;并且Pae能够明显下调LPS诱导的PI3K,p-PI3K,AKT,p-AKT的表达;下调NF-κB的表达水平并降低P65蛋白的生物活性,下调p-P65蛋白的表达;下调P65的转移入核。Pae对LPS损伤的VEC有保护作用,可能通过抑制PI3K/AKT及NF-κB信号通路相关信号蛋白的表达。

This paper was aimed to observe the anti-atheroslerosis effect of paeonol（ Pae） on the activation of PI3K/AKT-NF-κB and the proliferation activity of rat vasular endothelial cells induced by lipopolysaccharide（ LPS） and co-cultured with smooth muscle cells. Primary rat vascular endothelial cells（ VECs） and rat vascular smooth cells（ VSMCs） were cultured by predigesting and adhering tissue blocks. The VEC-VSMC co-culture model was established by Transwell chamber. LPS（ 100 μg·L^（-1）,7 h） was used to induce VEC injury. MTT assay were used to determine the VEC proliferation activity. Western blot was used to detect PI3 K / AKT and NF-κB＇s signaling pathways related protein expressions. The concentration of LPS-induced VECs injury was 100 μg · L^（-1）,and the time was 7 h. After the intervention on the above cell model for 24 h,paeonol（ 15,30,60 μmol·L^（-1）） could effectively inhibit LPSinduced VECs injury,block PI3 K / AKT-NF-κB signal transduction pathway thereby significantly affecting the proliferation of LPS-induced VECs co-cultured with SMCs. The anti-atherosclerosis mechanism of paeonol may be related to the reducing the inhibitory effect of the signaling pathway associated proteins of VEC PI3 K / AKT and NF-κB,thereby increasing the VEC livability under co-culture.