Zhang Qiong, Liu Mingfu, Nong Haibin, Zhang Yanan, Bai Yiguang, Liu Pan, Zong Shaohui, Zeng Gaofeng
School of Public Health of Guangxi Medical University, Nanning, China.
Department of Spine Osteopathia, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Front Pharmacol. 2022 Aug 22;13:925568. doi: 10.3389/fphar.2022.925568. eCollection 2022.
The death of spinal motor neurons (SMNs) after spinal cord injury (SCI) is a crucial cause, contributing to a permanent neurological deficit. Total flavonoids of hawthorn leaves (TFHL) have been confirmed to have potentially therapeutic for SCI. Nonetheless, the roles and mechanisms of TFHL in recovering neuromotor function and regenerating axons of SMNs have not been fully elucidated. In this study, TFHL was applied to treat rats with SCI and injured SMNs for 7 days. experiment, rats with SCI were evaluated by a BBB (Basso-Beattie-Bresnahan) score to assess their motor functional recovery. The morphology, microstructure, apoptosis, Nissl bodies, and autophagy of SMNs in spinal cord tissue were detected by Hematoxylin-eosin (HE) staining, transmission electron microscopy, TUNEL staining, Nissl staining, and immunohistochemistry respectively. experiment, the co-culture model of SMNs and astrocytes was constructed to simulate the internal environment around SMNs in the spinal cord tissue. The cell morphology, microstructure, axonal regeneration, and autophagy were observed via optical microscope, transmission electron microscopy, and immunofluorescence. The content of neurotrophic factors in the cell culture medium of the co-culture model was detected by ELISA. Moreover, the expression of axon-related and autophagy-related proteins in the spinal cord tissue and SMNs was measured by Western Blot. We demonstrated that TFHL improved the neuromotor function recovery in rats after SCI. We then found that TFHL significantly promoted injured spinal cord tissue repair, reduced apoptosis, and improved the functional status of neurons in spinal cord tissue . Meanwhile, the cell morphology, microstructure, and axonal regeneration of damaged SMNs also obviously were improved, and the secretion of neurotrophic factors was facilitated after treatment with TFHL . Further, we revealed that TFHL promoted autophagy and related protein expression and vitro. Taken together, our study suggested that TFHL might facilitate autophagy and have neuroprotective properties in SMNs to enhance the recovery of neuromotor function of rats with SCI.
脊髓损伤(SCI)后脊髓运动神经元(SMNs)死亡是导致永久性神经功能缺损的关键原因。山楂叶总黄酮(TFHL)已被证实对SCI具有潜在治疗作用。然而,TFHL在恢复神经运动功能和促进SMNs轴突再生中的作用及机制尚未完全阐明。本研究中,应用TFHL对SCI大鼠和损伤的SMNs进行7天治疗。实验中,通过Basso-Beattie-Bresnahan(BBB)评分评估SCI大鼠的运动功能恢复情况。分别采用苏木精-伊红(HE)染色、透射电子显微镜、TUNEL染色、尼氏染色和免疫组化检测脊髓组织中SMNs的形态、微观结构、凋亡、尼氏体和自噬情况。实验中,构建SMNs与星形胶质细胞的共培养模型以模拟脊髓组织中SMNs周围的内环境。通过光学显微镜、透射电子显微镜和免疫荧光观察细胞形态、微观结构、轴突再生和自噬情况。采用酶联免疫吸附测定(ELISA)检测共培养模型细胞培养基中神经营养因子的含量。此外,通过蛋白质免疫印迹法检测脊髓组织和SMNs中轴突相关蛋白和自噬相关蛋白的表达。我们证明TFHL可改善SCI大鼠的神经运动功能恢复。随后我们发现TFHL能显著促进损伤脊髓组织修复、减少细胞凋亡并改善脊髓组织中神经元的功能状态。同时,TFHL治疗后受损SMNs的细胞形态、微观结构和轴突再生也明显改善,且神经营养因子分泌增加。进一步研究发现,TFHL在体内和体外均能促进自噬及相关蛋白表达。综上所述,我们的研究表明TFHL可能通过促进自噬对SMNs具有神经保护作用,从而增强SCI大鼠神经运动功能的恢复。