Yang Yin, Guo Chunyang, Liao Bo, Cao Junjun, Liang Chen, He Xijing
Department of Orthopaedics, The Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
The Second Department of Orthopedics, Xi'an Central Hospital, Xi'an, Shaanxi 710003, P.R. China.
Int J Mol Med. 2017 Feb;39(2):423-429. doi: 10.3892/ijmm.2016.2838. Epub 2016 Dec 27.
Autophagy plays an important role in the progression of spinal cord injury (SCI). In this study, we aimed to examine the effects and potential mechanisms of action of BMP and activin membrane-bound inhibitor (BAMBI) in the progression of SCI. A rat model of SCI was established and the rats were injected with pLentiH1-BAMBI shRNA and pAd-BAMBI in the gray and white matter of the spinal cord at T8. After 14 days, motor function evaluation was measured according to the Basso Beattie Bresnahan (BBB) method and the number of motor neuron cell accounts in the anterior horns was measured by Nissl staining. The protein expression of levels light chain 3B (LC3B), Beclin 1, Bim and p62 were measured by western blot analysis. The concentrations of interleukin (IL)-1β, IL-6 and IL-10 were measured by ELISA. The results revealed that BAMBI expression was significantly decreased in the rats with SCI. The BBB scores and the number of motor neuron cell accounts in the anterior horns were significantly decreased in the pLentiH1-BAMBI shRNA injection group, and were increased in the pAd-BAMBI injection group, suggesting a neuroprotective effect of BAMBI in SCI. Moreover, the increased expression levels of LC3B and Beclin 1, and the decreased expression of Bim and p62 indicated that autophagy was significantly induced in the pAd-BAMBI injection group. Moreover, the overexpression of BAMBI also decreased the expression of transforming growth factor-β (TGF-β) and mammalian target of rapamycin (mTOR), and decreased the concentrations of IL-1β, IL-6 and IL-10. These results indicate that BAMBI plays a neuroprotective role by decreasing inflammation and activating autophagy in rats with SCI.
自噬在脊髓损伤(SCI)的进展中起重要作用。在本研究中,我们旨在探讨骨形态发生蛋白和激活素膜结合抑制剂(BAMBI)在SCI进展中的作用及潜在作用机制。建立大鼠SCI模型,并在T8脊髓灰质和白质中注射pLentiH1 - BAMBI shRNA和pAd - BAMBI。14天后,根据Basso Beattie Bresnahan(BBB)方法进行运动功能评估,并通过尼氏染色测量前角运动神经元细胞数量。通过蛋白质免疫印迹分析测量轻链3B(LC3B)、Beclin 1、Bim和p62的蛋白表达水平。通过酶联免疫吸附测定法测量白细胞介素(IL)-1β、IL - 6和IL - 10的浓度。结果显示,SCI大鼠中BAMBI表达显著降低。pLentiH1 - BAMBI shRNA注射组的BBB评分和前角运动神经元细胞数量显著降低,而pAd - BAMBI注射组则增加,表明BAMBI在SCI中具有神经保护作用。此外,pAd - BAMBI注射组中LC3B和Beclin 1表达水平升高,Bim和p62表达降低,表明自噬被显著诱导。此外,BAMBI的过表达还降低了转化生长因子-β(TGF - β)和雷帕霉素靶蛋白(mTOR)的表达,并降低了IL - 1β、IL - 6和IL - 10的浓度。这些结果表明,BAMBI通过减轻炎症和激活SCI大鼠的自噬发挥神经保护作用。