Li Si-Yuan, Fan Ting-Ting, Yin Jian, Wan Cai-Yun, Li Mei-Li, Xia Shuai-Shuai, Li Qiang, Li Liang
Provincial Key Laboratory of Traditional Chinese Medicine Diagnostics, Hunan University of Chinese Medicine, Changsha, 410208, China.
Key Laboratory of TCM Heart and Lung Syndrome Differentiation & Medicated Diet and Dietotherapy, Hunan University of Chinese Medicine, Changsha, 410208, China.
Chin J Integr Med. 2025 May 15. doi: 10.1007/s11655-025-4201-6.
OBJECTIVE: To investigate whether Buyang Huanwu Decoction (BYHWD) had a good curative effect on the neuroprotection of red nucleus neurons after spinal cord injury (SCI) and the possible molecular mechanism. METHODS: Ninety male Sprague-Dawley rats were divided into 5 groups (n=18 per group) according to a random number table, including the control, model, low- (12.78 g/kg, BL group), medium- (25.65 g/kg, BM group), and high-dose BYHWD groups (51.30 g/kg, BH group). A rubrospinal tract transection model in rats was established, and different doses of BYHWD were intragastrically administrated for 4 weeks. The forelimb locomotor function was recorded using the spontaneous vertical exploration test. Cyclic adenosine monophosphate (cAMP) level in red nucleus was detected through an enzyme-linked immunosorbent assay. The morphology and number of red nucleus neurons were observed using Nissl's staining and axonal retrograde tracing by Fluoro-Gold (FG). The expression of cAMP-dependent protein kinase A (PKA), nuclear factor kappa-B (NF-κB) p65, and brain-derived neurotrophic factor (BDNF) in red nucleus were detected using immunohistochemistry and quantitative real-time polymerase chain reaction. RESULTS: Compared with the control group, the utilization rate of bilateral forelimbs, unilateral right forelimbs, proportion of FG-labeled positive neurons, cAMP level, protein expressions of PKA and BDNF, and BDNF mRNA expression were significantly decreased in the model group (P<0.01), while NF-κB p65 was increased in the model group (P<0.01). Compared with the model group, the utilization rate of bilateral forelimbs and unilateral right forelimbs were significantly higher in the BL, BM and BH groups (P<0.01), the proportion of FG-labeled positive neurons, cAMP level, protein expressions of PKA and BDNF and BDNF mRNA expression in all BYHWD groups were increased (P<0.05 or P<0.01), while NF-κB p65 were decreased in all BYHWD groups (P<0.05 or P<0.01). CONCLUSIONS: BYHWD possesses a sound neuroprotective effect on red nucleus neurons after SCI, and the efficacy was dose-related. The mechanism may be related to regulating the cAMP/PKA/NF-κ B p65 signaling pathway, finally promoting expression of BDNF.
目的：探讨补阳还五汤（BYHWD）对脊髓损伤（SCI）后红核神经元的神经保护作用及可能的分子机制。 方法：将90只雄性Sprague-Dawley大鼠按随机数字表法分为5组（每组18只），包括对照组、模型组、补阳还五汤低剂量组（12.78 g/kg，BL组）、中剂量组（25.65 g/kg，BM组）和高剂量组（51.30 g/kg，BH组）。建立大鼠红核脊髓束横断模型，给予不同剂量补阳还五汤灌胃4周。采用自发垂直探索试验记录前肢运动功能。通过酶联免疫吸附测定法检测红核中环磷酸腺苷（cAMP）水平。采用尼氏染色和荧光金（FG）逆行轴突示踪法观察红核神经元的形态和数量。采用免疫组织化学和实时定量聚合酶链反应检测红核中cAMP依赖性蛋白激酶A（PKA）、核因子κB（NF-κB）p65和脑源性神经营养因子（BDNF）的表达。 结果：与对照组相比，模型组双侧前肢、单侧右前肢利用率，FG标记阳性神经元比例，cAMP水平，PKA和BDNF蛋白表达及BDNF mRNA表达均显著降低（P<0.01），而模型组NF-κB p65升高（P<0.01）。与模型组相比，BL、BM和BH组双侧前肢和单侧右前肢利用率显著升高（P<0.01），各补阳还五汤组FG标记阳性神经元比例、cAMP水平、PKA和BDNF蛋白表达及BDNF mRNA表达均升高（P<0.05或P<0.01），而各补阳还五汤组NF-κB p65降低（P<0.05或P<0.01）。 结论：补阳还五汤对SCI后红核神经元具有良好的神经保护作用，且疗效与剂量相关。其机制可能与调节cAMP/PKA/NF-κB p65信号通路，最终促进BDNF表达有关。
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