Wang Kun, Li Meng, Jin Linyu, Deng Chao, Chen Zhi, Chen Hao, Han Yingchao, Qian Lie, Li Xinfeng, Shen Hongxing
Department of Spine Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University 160 Pujian Rd. Shanghai 200127 China
Department of Ultrasound, Obstetrics and Gynecology Hospital, Fudan University Shanghai 200090 China.
RSC Adv. 2019 Oct 9;9(55):32072-32080. doi: 10.1039/c9ra04591k. eCollection 2019 Oct 7.
The present study was aimed at the investigation of the effects of melatonin on spinal cord injury (SCI) and the role of IGFBP3 in SCI both and . The rats received treatment with 100 mg kg melatonin or both melatonin and pGenesil-1-si-IGFBP3 (50 µg per g bw) after SCI surgery. The motor function in rats was measured using the Basso-Beattie-Bresnahan (BBB) scale score; perfusion vessel area was determined by injecting FITC-conjugated lycopersicon esculentum agglutinin lectin (FITC-LEA), whereas the blood-spinal cord barrier permeability was measured using Evans blue. The pericytes were isolated, and the cells were cultured under hypoxia, treated with melatonin or transfected with si-IGFBP3. RT-qPCR and western blotting were conducted for the determination of IGFBP3, VEGF, MMP-2, ICAM-1 and Ang1. The expression of IGFBP3 was significantly down-regulated in the SCI rats, and melatonin significantly enhanced the IGFBP3 level. Melatonin improved the motor function, reduced the neuron injury, and improved the microcirculation in rats. However, the down-regulation of IGFBP3 significantly reversed these effects. Moreover, in both the SCI rat spinal cord tissues and the pericytes under hypoxia, the expressions of IGFBP3 and Ang1 were significantly down-regulated, whereas those of the proteins MMP-2, VEGF and ICAM-1 were significantly up-regulated, and melatonin dramatically inhibited these changes. Melatonin could protect the rats from SCI by improving the microcirculation through the up-regulation of IGFBP3.
本研究旨在探讨褪黑素对脊髓损伤（SCI）的影响以及IGFBP3在SCI中的作用。大鼠在脊髓损伤手术后接受100mg/kg褪黑素或褪黑素与pGenesil-1-si-IGFBP3（每克体重50μg）联合治疗。采用Basso-Beattie-Bresnahan（BBB）量表评分测量大鼠的运动功能；通过注射异硫氰酸荧光素标记的番茄凝集素（FITC-LEA）测定灌注血管面积，而使用伊文思蓝测量血脊髓屏障通透性。分离周细胞，将细胞在缺氧条件下培养，用褪黑素处理或转染si-IGFBP3。进行RT-qPCR和蛋白质印迹法以测定IGFBP3、VEGF、MMP-2、ICAM-1和Ang1。IGFBP3在SCI大鼠中的表达显著下调，而褪黑素显著提高了IGFBP3水平。褪黑素改善了大鼠的运动功能，减少了神经元损伤，并改善了微循环。然而，IGFBP3的下调显著逆转了这些作用。此外，在SCI大鼠脊髓组织和缺氧条件下的周细胞中，IGFBP3和Ang1的表达均显著下调，而MMP-2、VEGF和ICAM-1蛋白的表达均显著上调，褪黑素显著抑制了这些变化。褪黑素可通过上调IGFBP3改善微循环来保护大鼠免受脊髓损伤。
