Li Weijuan, Chen Yaoqi, Li Songhai, Guo Xiaopeng, Zhou Wenping, Zeng Qiutang, Liao Yuhua, Wei Yumiao
Laboratory of Cardiovascular Immunology, Institute of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology Wuhan, Hubei 430022, China.
Am J Transl Res. 2014 Nov 22;6(6):678-90. eCollection 2014.
This study aimed to investigate the effects of agonistic antibody to angiotensin II type 1 receptor (AT1-AA) on atherosclerosis in male ApoE-/- mice which were employed to establish the animal models of AT1-AA in two ways. In the first group, mice were injected subcutaneously with conjugated AT1 peptide at multiple sites; in the second group, mice were infused with AT1-AA prepared from rabbits that were treated with AT1 peptide intraperitoneally. Mice in each group were further randomly divided into five subgroups and treated with AT1 peptide/AT1-AA, AT1 peptide/AT1-AA plus valsartan, AT1 peptide/AT1-AA plus fenofibrate, AT1 peptide/ AT1-AA plus pyrrolidine dithiocarbamate (PDTC) and control vehicle, respectively. Antibodies were detected in mice (except for mice in control group). Aortic atherosclerotic lesions were assessed by oil red O staining, while plasma CRP, TNF-α, nuclear factor-kappa B (NF-κB) and H2O2 were determined by ELISA. CCR2 (the receptor of MCP-1), macrophages, and smooth muscle cells were detected by immunohistochemistry. P47phox, MCP-1 and eNOS were detected by RT-PCR, while P47phox, NF-κB and MCP-1 were detected by Western blot assay. The aortic atherosclerotic lesions were significantly increased in AT1 peptide/AT1-AA treated mice, along with simultaneous increases in inflammatory parameters. However, mice treated with valsartan, fenofibrate or PDTC showed alleviated progression of atherosclerosis and reductions in inflammatory parameters. Thus, AT1-AA may accelerate aortic atherosclerosis in ApoE-/- mice, which is mediated, at least in part, by the inflammatory reaction involving nicotinamide-adenine dinucleotide phosphate oxidase, reactive oxygen species, and NF-κB. In addition, valsartan, fenofibrate and PDTC may inhibit the AT1-AA induced atherosclerosis.
本研究旨在探讨血管紧张素II 1型受体激动性抗体（AT1-AA）对雄性ApoE-/-小鼠动脉粥样硬化的影响，采用两种方法建立AT1-AA动物模型。第一组小鼠在多个部位皮下注射偶联的AT1肽；第二组小鼠输注由经腹腔注射AT1肽处理的兔子制备的AT1-AA。每组小鼠进一步随机分为五个亚组，分别用AT1肽/AT1-AA、AT1肽/AT1-AA加缬沙坦、AT1肽/AT1-AA加非诺贝特、AT1肽/AT1-AA加吡咯烷二硫代氨基甲酸盐（PDTC）和对照载体进行处理。在小鼠中检测到抗体（对照组小鼠除外）。通过油红O染色评估主动脉粥样硬化病变，同时通过ELISA测定血浆CRP、TNF-α、核因子-κB（NF-κB）和H2O2。通过免疫组织化学检测CCR2（MCP-1的受体）、巨噬细胞和平滑肌细胞。通过RT-PCR检测P47phox、MCP-1和eNOS，同时通过蛋白质免疫印迹法检测P47phox、NF-κB和MCP-1。经AT1肽/AT1-AA处理的小鼠主动脉粥样硬化病变显著增加，同时炎症参数也增加。然而，用缬沙坦、非诺贝特或PDTC处理的小鼠显示动脉粥样硬化进展减轻，炎症参数降低。因此，AT1-AA可能加速ApoE-/-小鼠的主动脉粥样硬化，这至少部分是由涉及烟酰胺腺嘌呤二核苷酸磷酸氧化酶、活性氧和NF-κB的炎症反应介导的。此外，缬沙坦、非诺贝特和PDTC可能抑制AT1-AA诱导的动脉粥样硬化。
