School of Traditional Chinese Medicine, Capital Medical University, Beijing, 100069, China.
Chin J Integr Med. 2014 Jan;20(1):24-30. doi: 10.1007/s11655-012-1236-2. Epub 2012 Dec 3.
To study the effects of Zuogui Pill (, ZGP) and Yougui Pill (, YGP) on the expressions of brain-derived neurotrophic factor (BDNF) and cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis (EAE), in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.
The rats were randomly divided into normal control (NC), model (MO), prednisone acetate (PA), ZGP and YGP groups. The EAE model of rat was established by injecting antigen containing myelin basic protein (MBP)68-86. The brain and spinal cord were harvested on the 14th and 28th day post-immunization (PI), the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction (PCR), and the cAMP levels were detected by using enzyme-immunoassay method.
(1) On the 28th day PI, the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated, especially in YGP group (P<0.05 or P<0.01). (2) On the 14th day PI, the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group (P<0.05 or P<0.01), and the cAMP level in YGP group was higher than that in ZGP group (P<0.05). The cAMP level in spinal cord also significantly increased in YGP group compared with MO, PA and ZGP groups, respectively (P<0.01). (3) On the 14th day PI, the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups, respectively (P<0.05). (4) On the 28th day PI, there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.
The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway, but the targets of molecular mechanism of ZGP may be different from those of YGP.
研究左归丸（ZGP）和右归丸（YGP）对实验性自身免疫性脑脊髓炎（EAE）大鼠脑源性神经营养因子（BDNF）和环磷酸腺苷（cAMP）/蛋白激酶 A（PKA）信号通路轴突再生表达的影响，以探讨 ZGP 和 YGP 促进轴突再生的可能机制。
将大鼠随机分为正常对照组（NC）、模型组（MO）、醋酸泼尼松龙组（PA）、ZGP 组和 YGP 组。用髓鞘碱性蛋白（MBP）68-86 抗原免疫大鼠建立 EAE 模型。免疫后第 14 天和第 28 天取脑和脊髓，用 Western blot 分析和实时定量聚合酶链反应（PCR）检测大鼠脑和脊髓中 BDNF 和 PKA 的蛋白和 mRNA 表达，用酶联免疫吸附法检测 cAMP 水平。
（1）PI 第 28 天，ZGP 和 YGP 组大鼠脑白质和脊髓中 BDNF 的 mRNA 表达上调，YGP 组尤为明显（P<0.05 或 P<0.01）。（2）PI 第 14 天，PA 和 YGP 组大鼠脑白质中 cAMP 水平明显高于 MO 组（P<0.05 或 P<0.01），YGP 组高于 ZGP 组（P<0.05）。YGP 组大鼠脊髓中 cAMP 水平也明显高于 MO、PA 和 ZGP 组（P<0.01）。（3）PI 第 14 天，ZGP 组大鼠脊髓中 PKA 表达明显低于 MO 和 YGP 组（P<0.05）。（4）PI 第 28 天，YGP 大鼠脑白质中 cAMP 与 PKA 表达呈正相关。
结果提示 ZGP 和 YGP 可能通过调节 cAMP/PKA 信号转导通路促进轴突再生，但 ZGP 的分子机制靶点可能与 YGP 不同。
