Zhang Chunyu, Huang Wentao, Huang Chen, Zhou Chengqian, Tang Yukuan, Wei Wei, Li Yongsheng, Tang Yukuan, Luo Yu, Zhou Quan, Chen Wenli
MOE Key Laboratory of Laser Life Science, Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China.
Front Bioeng Biotechnol. 2022 Feb 28;10:821256. doi: 10.3389/fbioe.2022.821256. eCollection 2022.
Magnetic resonance imaging (MRI) could be the ideal diagnostic modality for early rheumatoid arthritis (RA). Vascular cell adhesion molecule-1 (VCAM-1) is highly expressed in synovial locations in patients with RA, which could be a potential target protein for RA diagnosis. The peptide VHPKQHR (VHP) has a high affinity to VCAM-1. To make the contrast agent to target RA at an early stage, we used VHP and ultrasmall paramagnetic iron oxide (USPIO) to synthesize UVHP (U stands for USPIO) through a chemical reaction with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. The size of UVHP was 6.7 nm; the potential was -27.7 mV, and the / value was 1.73. Cytotoxicity assay exhibited that the cell survival rate was higher than 80% at even high concentrations of UVHP (Fe concentration 200 µg/mL), which showed the UVHP has low toxicity. Compared with no TNF-α stimulation, VCAM-1 expression was increased nearly 3-fold when mouse aortic endothelial cells (MAECs) were stimulated with 50 ng/mL TNF-α; cellular Fe uptake was increased very significantly with increasing UVHP concentration under TNF-α treatment; cellular Fe content was 17 times higher under UVHP with Fe concentration 200 µg/mL treating MAECs. These results indicate that UVHP can target overexpression of VCAM-1 at the cellular level. RA mice models were constructed with adjuvant-induced arthritis. MRI and biodistribution results show that the signal intensity of knee joints was increased significantly and Fe accumulation in RA model mice compared with normal wild-type mice after injecting UVHP 24 h. These results suggest that we have synthesized a simple, low-cost, and less toxic contrast agent UVHP, which targeted VCAM-1 for early-stage RA diagnosis and generates high contrast in T-weighted MRI.
磁共振成像（MRI）可能是早期类风湿性关节炎（RA）的理想诊断方式。血管细胞黏附分子-1（VCAM-1）在RA患者的滑膜部位高表达，这可能是RA诊断的潜在靶蛋白。肽VHPKQHR（VHP）对VCAM-1具有高亲和力。为了使造影剂在早期靶向RA，我们使用VHP和超小顺磁性氧化铁（USPIO）通过与1-（3-二甲基氨基丙基）-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺的化学反应合成了UVHP（U代表USPIO）。UVHP的尺寸为6.7纳米；电位为-27.7毫伏，ζ/电位值为1.73。细胞毒性试验表明，即使在高浓度的UVHP（铁浓度为200微克/毫升）下，细胞存活率仍高于80%，这表明UVHP具有低毒性。与未用肿瘤坏死因子-α（TNF-α）刺激相比，当用50纳克/毫升TNF-α刺激小鼠主动脉内皮细胞（MAECs）时，VCAM-1表达增加了近3倍；在TNF-α处理下，随着UVHP浓度的增加，细胞铁摄取显著增加；在铁浓度为200微克/毫升的UVHP处理MAECs时，细胞铁含量高出17倍。这些结果表明，UVHP可以在细胞水平上靶向VCAM-1的过表达。用佐剂诱导的关节炎构建RA小鼠模型。MRI和生物分布结果表明，注射UVHP 24小时后，与正常野生型小鼠相比，RA模型小鼠膝关节的信号强度显著增加，铁蓄积。这些结果表明，我们合成了一种简单、低成本且毒性较小的造影剂UVHP，其靶向VCAM-1用于早期RA诊断，并在T加权MRI中产生高对比度。
