Tu Zhi-Lan, Yu Bo, Huang Dong-Ya, Ojha Rajeev, Zhou Shu-Kui, An He-Di, Liu Rong, Du Cui, Shen Nan, Fu Jian-Hui, Hou Shuang-Xing
Department of Neurology, East Hospital, Tongji University School of Medicine, Shanghai 200120, P.R. China.
Department of Neurology, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai 201399, P.R. China.
Mol Med Rep. 2017 Sep;16(3):2347-2354. doi: 10.3892/mmr.2017.6869. Epub 2017 Jun 28.
The present study aimed to investigate protein expression levels of intra‑ and extracranial atherosclerosis in rabbits following administration of a high‑fat diet. Rabbits were randomly divided into control (group A; n=9) and high‑fat diet (group B; n=9) groups. At week 12, tissues were sectioned from the common carotid artery (CCA) and middle cerebral artery (MCA). Pathological analysis was performed. Differential protein expression levels were examined by 2‑D gel electrophoresis (2‑DE) and mass spectrometry (MS) analysis and validated by western blotting. Serum lipid levels, the intima‑media thickness (IMT) and degree of atherosclerosis of the CCA and MCA were increased at week 12 in the high‑fat diet group compared with rabbits that received a normal diet. 2‑DE and MS analysis of the protein extracted from CCA and MCA detected >439 different proteins; the expression of 25 proteins was altered, and 8 proteins [albumin A chain, tropomyosin α‑1 chain (TPM1), heat shock protein 70 (HSP70), α‑smooth muscle actin, β‑galactose binding agglutinin, TPM4 isoform 2, cell keratin 9, single octylic acid glyceride β‑2) demonstrated significant alterations in expression levels. Due to limited antibody sources, only three differentially expressed proteins (TPM1, HSP70 and α‑smooth muscle actin) were examined by western blotting. The results of our previous study demonstrated that hyperlipidemia affected the IMT of intracranial and extracranial cerebral arteries. In the present study, protein expression levels of TPM1 and α‑smooth muscle actin from extracranial cerebral arteries were significantly increased compared with intracranial cerebral arteries; however, protein expression levels of HSP70 from intracranial cerebral arteries was increased compared with extracranial cerebral arteries. The differences may be closely associated with cell proliferation and metastasis, and oxidoreduction, in intra‑ and extracranial cerebral atherosclerosis. HSP70 may have protective properties against atherosclerosis via underlying anti‑inflammatory mechanisms, furthermore, differential protein expression levels (TPM1, HSP70 and α‑smooth muscle actin) between intra‑ and extracranial cerebral arteries may facilitate the identification of novel biological markers for the diagnosis and treatment of cerebral arteriosclerosis.
本研究旨在探讨高脂饮食喂养的家兔颅内和颅外动脉粥样硬化的蛋白质表达水平。将家兔随机分为对照组(A组,n = 9)和高脂饮食组(B组,n = 9)。在第12周时,从颈总动脉(CCA)和大脑中动脉(MCA)取材切片并进行病理分析。通过二维凝胶电泳(2-DE)和质谱(MS)分析检测差异蛋白质表达水平,并通过蛋白质印迹法进行验证。与正常饮食的家兔相比,高脂饮食组家兔在第12周时血清脂质水平、CCA和MCA的内膜中层厚度(IMT)及动脉粥样硬化程度均升高。对从CCA和MCA提取的蛋白质进行2-DE和MS分析,检测到>439种不同蛋白质;25种蛋白质的表达发生改变,8种蛋白质[白蛋白A链、原肌球蛋白α-1链(TPM1)、热休克蛋白70(HSP70)、α-平滑肌肌动蛋白、β-半乳糖结合凝集素、TPM4亚型2、细胞角蛋白9、单辛酸甘油酯β-2]的表达水平有显著变化。由于抗体来源有限,仅通过蛋白质印迹法检测了三种差异表达蛋白质(TPM1、HSP70和α-平滑肌肌动蛋白)。我们之前的研究结果表明,高脂血症会影响颅内和颅外脑动脉的IMT。在本研究中,与颅内脑动脉相比,颅外脑动脉中TPM1和α-平滑肌肌动蛋白的蛋白质表达水平显著升高;然而,与颅外脑动脉相比,颅内脑动脉中HSP70的蛋白质表达水平升高。这些差异可能与颅内和颅外脑动脉粥样硬化中的细胞增殖和转移以及氧化还原密切相关。HSP70可能通过潜在的抗炎机制对动脉粥样硬化具有保护作用,此外,颅内和颅外脑动脉之间差异蛋白质表达水平(TPM1、HSP70和α-平滑肌肌动蛋白)可能有助于识别用于诊断和治疗脑动脉硬化的新型生物标志物。