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ABSTRACT
Objectives
In this study, we aimed to verify the neuroprotective effects of pycnogenol (PYC) on spinal cord injury (SCI) and to determine the underlying mechanisms.
Methods
Male Wistar rats were selected to establish a model of SCI in accordance with the Allen’s protocol. The rats in the PYC group were treated with 100 mg/kg PYC by intraperitoneal injection 15 minutes after SCI. The Basso, Beattie and Bresnahan (BBB) scale was used to evaluate locomotor activity. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) production were detected by ELISA. The expression of Cleaved-caspase 3, Bcl-2, Bax and the levels of Cytochrome c (Cyt-c) were analysed by Western blot or Immunohistochemistry. Furthermore, we used the JC-1 fluorescent probe to analyse the mitochondrial membrane potential (ΔΨm).
Results
The rats that received PYC had significantly higher BBB scores than the control lesion rats. PYC treatment resulted in reduced bleeding in spinal cord tissue and proliferation of glial cells, greater numbers of anterior horn neurons, more complete structures of residual neurons, and significant improvement in Nissl body morphology. In addition, PYC reduced MDA production and increased SOD activity. The mitochondrial membrane potential (MMP) was significantly increased in the PYC treatment group compared with the SCI group. In addition, PYC decreased the expression of Cleaved-caspase 3 and Bax and the release of Cyt-c and increased the expression of Bcl-2 in the SCI rats.
Conclusions
The above findings suggested that PYC can improve motor function and reduce neuronal apoptosis after SCI by stabilizing the MMP through the inhibition of oxidative stress.
Abbreviations
DMSO: dimethyl sulfoxide; IHC: immunological histological chemistry; MDA: malondialdehyde; PBS: phosphate buffered saline; PMSF: phenylmethanesulfonyl fluoride; PVDF: polyvinylidene difluoride; PYC: Pycnogenol; RIPA: radio immunoprecipitation assay; SCI: spinal cord injury; SOD: superoxide dismutase
Acknowledgments
In this study, author Lin Tao designed the experiment and review the manuscript, Xuan Liu carried on the most experiment process and Wacili Da participated in some process and write the manuscript. In addition, Zhengbo Tao makes a contribution in graph editing, cell culture, literature research and other works. Yue Zhu is responsible for the work and lead all authors to complete the whole subject.
Disclosure statement
No potential conflict of interest was reported by the authors.
Additional information
Funding
Notes on contributors
Lin Tao
Lin Tao is an orthopedic residents and instructor at the First Affiliated Hospital of China Medical University, with a Ph.D. degree. He has carried out research in the area of “osteoporosis、spinal cord injury and osteosarcoma”, and published several papers in academic journals.
Xuan Liu
Xuan Liu Master of Medicine, is an orthopedic resident, currently working at Department of the Orthopedic of The Affiliated Hospital of Chengdu University. He graduated from China Medical University,Shenyang, Liaoning, China. His research interests are focused on spinal cord injury and drug discovery.
Wacili Da
Wacili Da is a PhD Candidate in orthopedics, at the China Medical University. His research interests include the investigation of the pathogenesis of osteoporosis 、osteosarcoma and the recovery of spinal cord injury.
Zhengbo Tao
Zhengbo Tao is a Master degree candidate at the Department of orthopedics of the China Medical University. He has graduated from The Second Military Medical University, Shanghai, China. His research interests focus on osteoporosis、spinal cord injury and bibliometrics.
Yue Zhu
Yue Zhu is a clinician and academic specialized in osteoporosis, spinal cord injury and scoliosis, with a Ph.D. degree. He is the vice president and director of the departments of orthopedics, at the First Affiliated Hospital of China Medical University. He has published more than 70 papers in peer reviewed journals. He has been awarded several times for his clinical and scientific work.