目的:观察天麻素结合电针对局灶性脑缺血大鼠尾壳核生长相关蛋白43(growth-associated protein, GAP-43)和突触素(synaptophysin, SYP)蛋白表达是否具有协同增效作用,探讨针药结合治疗脑缺血的部分神经生物学机制。方法:48只雄性成年Sprague-Dawley(SD)大鼠,随机分为正常组、假手术组、模型组、天麻素组、电针组、天麻素+电针组。采用右侧大脑中动脉栓塞(middle cerebral artery occlusion, MCAO)法复制FCI模型。造模成功后2周,天麻素组、电针组和天麻素+电针组大鼠分别给予天麻素注射液、电针及天麻素结合电针连续治疗2周。采用免疫组织化学法检测大鼠尾壳核中GAP-43和SYP蛋白的表达情况。结果:各组大鼠尾壳核中均有GAP-43和SYP蛋白表达。与正常组比较,假手术组大鼠尾壳核中GAP-43和SYP蛋白的表达差异无统计学意义(P＞0.05);模型组GAP-43蛋白的表达增加,SYP蛋白的表达减少,差异均有统计学意义(P＜0.05)。与模型组比较,天麻素组、电针组和天麻素+电针组大鼠尾壳核中GAP-43和SYP蛋白的表达均增加,差异有统计学意义(P＜0.05)。天麻素+电针组大鼠尾壳核中GAP-43和SYP蛋白的表达较天麻素组、电针组增加,差异有统计学意义(P＜0.05)。结论:天麻素结合电针可上调局灶性脑缺血大鼠尾壳核中GAP-43和SYP蛋白的表达,这可能有助于促进神经元损伤的修复,且针药结合治疗的效果优于单独天麻素和电针治疗。
Objective: To explore the neurobiological mechanism of acupuncture combined with medicine in the treatment of cerebral ischemia by observing the synergistic effect of gastrodin combining with electro-acupuncture (EA) on the expression of growth associated protein 43(GAP-43) and synaptophysin (SYP) in caudate putamen nucleus of rats with focal cerebral ischemia (FCI). Methods: A total of 48 Sprague-Dawley (SD) rats were randomly divided into 6 groups (normal group, sham operation group, model group, gastrodin group, EA group and gastrodin+EA group). The model was prepared by middle cerebral artery occlusion (MCAO). 2 weeks after modeling, rats were given gastrodin, EA, or gastrodin+EA for continuous treatment in 2 successive weeks. Then expressions of GAP-43 and SYP protein in the caudate putamen nucleus were detected by immunohistochemistry. Results: Positive expression of GAP-43 and SYP protein was found in each group of rats. Expressions of GAP-43 in model group was higher than the normal group and sham operation group (P＜0.05), expressions of SYP in model group was lower than the normal group and sham operation group (P＜0.05). Compared with the model group, the positive expressions of GAP-43 and SYP in gastrodin group, the EA group, and the gastrodin+EA group increased significantly (P＜0.05). The positive expressions of GAP-43 and SYP increased obviously in the gastrodin+EA group compared to those of the EA group and the medication group (P＜0.05). Conclusion: Gastrodin administration and EA intervention can up-regulated the expressions of GAP-43 and SYP protein in the caudate putamen nucleus of FCI rats, which may contribute action in promoting the repair of injured neurons. The effect of gastrodin+EA is better than simple gastrodin or EA treatment.
[1] Li HB, Wu F, Miao HC, et al. Effects of polysaccharide of gastrodia elata blume and electro-acupuncture on expressions of brain-derived neurotrophic factor and stem cell factor protein in caudate putamen of focal cerebral ischemia rats[J].Med Sci Monit Basic Res, 2016, 22: 175-180.
[2] 董江宁, 熊克仁, 王术生, 等. 局灶性脑缺血大鼠尾壳核CT灌注成像与NOS表达研究[J]. 实用放射学杂志, 2008, (6): 840-842+849.
[3] Okada M, Kawagoe Y, Takasugi T, et al. JNK1-Dependent Phosphorylation of GAP-43 Serine 142 is a Novel Molecular Marker for Axonal Growth[J]. Neurochem Res, 2022, 47(9): 2668-2682.
[4] Titlow J, Robertson F, Järvelin A, et al. Syncrip/hnRNP Q is required for activity-induced Msp300/Nesprin-1 expression and new synapse formation[J]. J Cell Biol, 2020, 219(3): e201903135.
[5] Qiu CW, Liu ZY, Zhang FL, et al. Post-stroke gastrodin treatment ameliorates ischemic injury and increases neurogenesis and restores the Wnt/β-Catenin signaling in focal cerebral ischemia in mice[J]. Brain Res, 2019, 1712: 7-15.
[6] 陈岚榕, 王林林, 陈松, 等. 艾灸曲池、合谷穴结合康复训练对脑梗死恢复期患者上肢功能障碍恢复疗效的回顾性分析[J]. 中华中医药杂志, 2017, 32(12): 5652-5655.
[7] Hermann DM, Popa-Wagner A, Kleinschnitz C, et al. Animal models of ischemic stroke and their impact on drug discovery[J]. Expert Opin Drug Discov, 2019, 14(3): 315-326.
[8] Chung D, Shum A, Caraveo G. GAP-43 and BASP1 in axon regeneration: implications for the treatment of neurodegenerative diseases[J]. Front Cell Dev Biol, 2020, 8: 567537.
[9] Lin JB, Hao CZ, Gong Y, et al. Effect of tetramethylpyrazine on neuroplasticity after transient focal cerebral ischemia reperfusion in rats[J].Evid Based Complement Alternat Med, 2021, 2021: 1587241.
[10] 向杜炼, 唐银杉, 邵淑君, 等. 电针治疗对APP/PS1小鼠海马区突触相关蛋白SYP、PSD95表达的影响[J]. 中华中医药杂志, 2018, 33(11): 5143-5146.
[11] 惠建荣, 羊璞, 赵娴, 等. 电针结合传统针刺百会、大椎穴对脑缺血模型大鼠GAP-43表达的影响[J]. 陕西中医, 2020, 41(7): 856-859.
[12] Liu Y, Gao JL, Peng M, et al. A review on central nervous system effects of gastrodin[J].Front Pharmacol, 2018, 9: 24.
[13] Li SS, Hua XY, Zheng MX, et al. Electroacupuncture treatment improves motor function and neurological outcomes after cerebral ischemia/reperfusion injury[J].Neural Regen Res, 2022, 17(7): 1545-1555.
[14] 崔晶晶, 高俊虹, 王玉敏, 等. 针药结合增效机制研究的新思路探讨[J]. 针刺研究, 2010, 35(2): 146-150.
[15] Shi L, Cao HM, Li Y, et al. Electroacupuncture improves neurovascular unit reconstruction by promoting collateral circulation and angiogenesis[J]. Neural Regen Res, 2017, 12(12): 2000-2006.
[16] 魏彦珍, 刘轲, 张怀亮. 针药结合治疗脑梗死机制研究进展[J]. 辽宁中医药大学学报, 2014, 16(02): 217-219.
