Aim This study was to evaluate the effect of arsenic trioxide （As2O3） on the transgenic TNF-α promoter activity in cultured vascular smooth muscle cells （VSMCs） and THP-1 monocytes. Methods Human TNF-α promoter was constructed by reporter gene system and was transiently transfected into VSMCs and THP-1 in vitro. The promoter activity was tested by luciferase activity with or without LPS and Ang Ⅱ stimulation, before and after different dosage of As2O3 treatment. Results 1. TNF-α promoter effectively expressed in VSMCs and THP-1 compared with CMV promoter （58.3% and 80.9%, respectively）. Both LPS and Ang Ⅱ significantly up-regulated TNF-α promoter activity （P〈0.05）. 2. As2O3 significantly inhibited, both intact and LPS/Ang Ⅱ stimulated promoter activity, in a dose dependent manner （P〈0.05）, and in both cell type. Conclusion These results manifested that, the inhibition effect of As2O3 on the activity of human TNF-α promoter indicated its potential inhibition on pro-inflammatory cytokine genes expression at transcriptional level and its potential anti-inflammatory property in the cardiovascular system.
目的观察国产血管紧张素1型受体拮抗剂厄贝沙坦抑制兔颈动脉球囊损伤后再狭窄(RS)的作用,探讨 RS 可能的机理。方法将48只大耳白兔随机分为实验7、14、28 d 组并各设对照组(每组8只),以球囊损伤左颈总动脉,建立再狭窄模型。术后7、14、28 d 不同时点取材。实验组术前3 d 喂饲厄贝沙坦(35 mg/kg)至处死。病理 HE 染色观察动脉壁内、中膜面积、管腔面积;免疫组化法测定核因子-κB(NF-κB)、核增殖抗原(PCNA)核易位阳性细胞率;实时荧光定量聚合酶链反应(PCR)测定κB抑制蛋白(I-κB)、单核细胞趋化蛋白 mRNA 表达。结果厄贝沙坦实验组与对照组相比,术后7 d 时明显抑制内膜的增生(0.34 mm^2±0.15 mm^2 vs 1.05 mm^2±0.38 mm^2,P<0.05),28 d 时管腔面积明显较大(4.25 mm^2±0.29 mm^2 vs 2.56 mm^2±1.02 mm^2,P<0.05)。术后7 d 时厄贝沙坦明显抑制 NF-κB p65核易位(P<0.05)和 PCNA 的表达(P<0.05)。术后7 d 时厄贝沙坦实验组 I-κB mRNA 含量明显较低(7.2拷贝/μl±0.9拷贝/μl vs 15.6拷贝/μl±0.7拷贝/μl,P<0.05),厄贝沙坦实验组在各时相点均明显抑制 MCP-1 mRNA 的表达(P<0.05)。结论厄贝沙坦通过抑制NF-κB的激活而调控 MCP-1、PCNA 的表达,有效抑制再狭窄的发生发展。
