Zhang Zhen, Dai Min
Zhongguo Zhong Yao Za Zhi. 2014 Mar;39(6):1058-63.
To observe the changes in the adhesive function of vascular endothelial cells (VEC) and rat monocytes induced by lipopolysaccharide (LPS) and co-cultured with smooth muscle cells (SMC) and the intervention effect of paeonol (Pae).
Primary rat vascular endothelial cells (VECs) and rat vascular smooth muscle cells (VSMCs) were cultured by predigesting and adhering tissue blocks. The VEC-VSMC co-culture model was established by Transwell chamber. LPS was used to induce VEC injury. MTT assay and LDH assay were used to determine the VEC activity. ELISA assay was used to detect IL-1beta and TNF-alpha secreted by the VEC. The immunocytochemistry assay was carried out to detect the expression of ICAM-1. The Rose Bengal Staining was used to test adhesive function between VECs and monocytes.
The concentration of LPS-induced VEC injury was 100 microg x L(-1), and the time was 7 h. after the intervention on the above cell model for 24 h, Paeonol (15, 30, 60 micromol x L(-1)) could effectively inhibit LPS-induced VEC injury and VEC injury, significantly enhance the survival rate of LPS-injured VECs, decrease IL-1beta and TNF-alpha secreted by the injured VEC, and reduce the expression of ICAM-1, so as to inhibit the adhesion of LPS-induced VECs and monocytes.
Paeonol could inhibit IL-1beta and TNF-alpha expression to protect VECs from being injured by LPS, and reduce ICAM-1 expression to inhibit the adhesion between VECs and monocytes.
观察脂多糖(LPS)诱导的血管内皮细胞(VEC)与大鼠单核细胞黏附功能变化以及与平滑肌细胞(SMC)共培养时丹皮酚(Pae)的干预作用。
采用组织块消化贴壁法培养原代大鼠血管内皮细胞(VECs)和大鼠血管平滑肌细胞(VSMCs)。通过Transwell小室建立VEC-VSMC共培养模型。用LPS诱导VEC损伤。采用MTT法和LDH法检测VEC活性。用ELISA法检测VEC分泌的IL-1β和TNF-α。进行免疫细胞化学检测ICAM-1的表达。用孟加拉玫瑰红染色检测VEC与单核细胞之间的黏附功能。
LPS诱导VEC损伤的浓度为100μg·L⁻¹,时间为7 h。对上述细胞模型干预24 h后,丹皮酚(15、30、60μmol·L⁻¹)可有效抑制LPS诱导的VEC损伤,显著提高LPS损伤VEC的存活率,降低损伤VEC分泌的IL-1β和TNF-α,减少ICAM-1的表达,从而抑制LPS诱导的VEC与单核细胞的黏附。
丹皮酚可抑制IL-1β和TNF-α表达,保护VEC免受LPS损伤,降低ICAM-1表达,抑制VEC与单核细胞之间的黏附。