Zhantao Liu1, Xuying Ren2, Zhihong Yang1, Yongjuan Zhao1, Lixia Ji1, Jing Li1, Wang Yue1. 1. Department of Pharmacy, Medical College of Qingdao University Qingdao, Shandong 266021, China. 2. Department of Pharmacy, Medical College of Qingdao University Qingdao, Shandong 266021, China ; Hospital of Laigang Group Co. Ltd. Laiwu, Shandong 271126, China.
Abstract
PURPOSE: Indol-2,3-dione (ISA), a natural substance with clear structure, has been shown to process anti-inflammatory, antioxidant and anti-atherosclerosis activity in vivo. The purpose of this study was to investigate the effect and mechanism of ISA on AS by primary rat cardiac microvascular endothelial cells (rCMEC). METHODS: rCMEC cells were primary cultured, appraised by cell morphology and immune cell chemical dyeing, and passaged to the 3(rd) generation. The effect of ISA on the activity of rCMEC induced by oxidized low-density lipoprotein (ox-LDL) was detected by MTT method. Then we studied the effect of ISA on the adhesion of monocyte with rCMEC induced by ox-LDL. The secretion of interleutin-6 (IL-6) and tumor necrosis factor-α (TNF-α) by rCMEC were measured by enzyme-linked immunosorbent assay (ELISA) method. Finally the mRNA level of IL-6 and TNF-α of rCMEC were analyzed by real time RT-PCR. Results MTT result indicated that ISA (10(-8)-10(-6) g/L) could inhibit rCMEC injury induced by ox-LDL in a dose-dependent manner (P < 0.05). The adhesion of monocyte with rCMEC induced by ox-LDL was inhibited by ISA in a dose-dependent manner (P < 0.05). The levels of IL-6 and TNF-α of ISA groups were significantly decreased in a dose-dependent manner compared with model group (P < 0.05). The mRNA expressions of IL-6 and TNF-α of ISA groups were also downregulated significantly compared with model group (P < 0.05). Conclusions ISA can prevent atherosclerotic lesion. Its mechanism may be that it can defend against the oxidation damage to rCMEC, inhibit the adhesion of monocyte to rCMEC, and reduce inflammatory secretions of IL-6 and TNF-α.
PURPOSE:Indol-2,3-dione (ISA), a natural substance with clear structure, has been shown to process anti-inflammatory, antioxidant and anti-atherosclerosis activity in vivo. The purpose of this study was to investigate the effect and mechanism of ISA on AS by primary rat cardiac microvascular endothelial cells (rCMEC). METHODS: rCMEC cells were primary cultured, appraised by cell morphology and immune cell chemical dyeing, and passaged to the 3(rd) generation. The effect of ISA on the activity of rCMEC induced by oxidized low-density lipoprotein (ox-LDL) was detected by MTT method. Then we studied the effect of ISA on the adhesion of monocyte with rCMEC induced by ox-LDL. The secretion of interleutin-6 (IL-6) and tumor necrosis factor-α (TNF-α) by rCMEC were measured by enzyme-linked immunosorbent assay (ELISA) method. Finally the mRNA level of IL-6 and TNF-α of rCMEC were analyzed by real time RT-PCR. Results MTT result indicated that ISA (10(-8)-10(-6) g/L) could inhibit rCMEC injury induced by ox-LDL in a dose-dependent manner (P < 0.05). The adhesion of monocyte with rCMEC induced by ox-LDL was inhibited by ISA in a dose-dependent manner (P < 0.05). The levels of IL-6 and TNF-α of ISA groups were significantly decreased in a dose-dependent manner compared with model group (P < 0.05). The mRNA expressions of IL-6 and TNF-α of ISA groups were also downregulated significantly compared with model group (P < 0.05). Conclusions ISA can prevent atherosclerotic lesion. Its mechanism may be that it can defend against the oxidation damage to rCMEC, inhibit the adhesion of monocyte to rCMEC, and reduce inflammatory secretions of IL-6 and TNF-α.