Wang Yongrui, Zheng Xianli, Yan Xingke
School of Acupuncture-Moxibustion and Tuina, Gansu University of CM, Lanzhou 730000, China.
Zhongguo Zhen Jiu. 2024 Nov 12;44(11):1281-8. doi: 10.13703/j.0255-2930.20231116-k0006.
To observe the effect of acupuncture (acupuncture for soothing the liver and regulating the spirit) on the protein kinase C/extracellular signal-regulated kinase/cAMP response element-binding protein (PKC/ERK/CREB) signaling pathway in the bed nucleus of the stria terminalis (BNST) of rats with post-traumatic stress disorder (PTSD), and to explore the mechanism of acupuncture on alleviating anxiety and fear in PTSD.
Fifty SPF-grade male SD rats were randomly divided into a blank group (10 rats) and a PTSD model group (40 rats). The PTSD model was induced by using a combination of closed electric shock and forced exhaustive swimming. Thirty successfully modeled rats were randomly assigned to a model group, a medication group, and an acupuncture group, with 10 rats in each group. The rats in the medication group were treated with paroxetine hydrochloride solution by gavage, once daily for 12 consecutive days. The rats in the acupuncture group were treated with acupuncture at "Baihui" (GV 20) and bilateral "Neiguan" (PC 6), "Shenmen" (HT 7), "Taichong" (LR 3). "Baihui" (GV 20) was needled daily, while the other acupoints were alternately needled on the left side on odd days and the right side on even days, once daily for 12 consecutive days. Anxiety and fear behaviors changes were assessed by using the open field test and elevated plus maze test. Histological changes in the BNST were observed by using HE staining and Nissl staining. The expression of PKC, phosphorylated PKC (p-PKC), ERK1/2, phosphorylated ERK1/2 (p-ERK1/2), and p-CREB proteins in the BNST were detected by using Western blot.
Compared with the blank group, the model group showed decreased time and total distance spent in the center of the open field and on the open arms of the elevated plus maze (<0.05); the BNST tissues in the model group exhibited a reduced number of neurons, disorganized cell arrangement, cell shrinkage, nuclear condensation, abnormal neuronal structure, uneven Nissl staining, and reduced Nissl bodies. The model group showed increased protein expression of p-PKC and p-PKC/PKC ratio (<0.05) and decreased protein expression of p-ERK1/2, p-CREB, and p-ERK1/2/ERK1/2 ratio (<0.05). Compared with the model group, the medication group and the acupuncture group showed increased time and total distance spent in the center of the open field and on the open arms of the elevated plus maze (<0.05); the BNST tissues showed increased number of neurons, more organized cell arrangement, improved neuronal structure, and increased Nissl bodies; the medication group and the acupuncture group also showed decreased p-PKC protein expression and p-PKC/PKC ratio (<0.05) and increased p-ERK1/2, p-CREB protein expression, and p-ERK1/2/ERK1/2 ratio (<0.05).
acupuncture could alleviate anxiety and fear behaviors in PTSD rats, and improve neuronal damage in the BNST. The mechanism may be related to the regulation of the PKC/ERK/CREB signaling pathway in the BNST.
观察针刺（疏肝调神针法）对创伤后应激障碍（PTSD）大鼠终纹床核（BNST）中蛋白激酶C/细胞外信号调节激酶/cAMP反应元件结合蛋白（PKC/ERK/CREB）信号通路的影响，探讨针刺缓解PTSD大鼠焦虑恐惧的作用机制。
将50只SPF级雄性SD大鼠随机分为空白组（10只）和PTSD模型组（40只）。采用封闭电击结合强迫性 exhaustive游泳的方法诱导PTSD模型。将30只成功造模的大鼠随机分为模型组、药物组和针刺组，每组10只。药物组大鼠采用盐酸帕罗西汀溶液灌胃治疗，每日1次，连续12天。针刺组大鼠针刺“百会”（GV 20）及双侧“内关”（PC 6）、“神门”（HT 7）、“太冲”（LR 3）。“百会”（GV 20）每日针刺，其他穴位隔日交替针刺，左侧穴位在奇数日针刺，右侧穴位在偶数日针刺），每日1次，连续12天。采用旷场试验和高架十字迷宫试验评估焦虑和恐惧行为变化。采用HE染色和尼氏染色观察BNST的组织学变化。采用蛋白质印迹法检测BNST中PKC、磷酸化PKC（p-PKC）、ERK1/2、磷酸化ERK1/2（p-ERK1/2）和p-CREB蛋白的表达。
与空白组比较，模型组大鼠在旷场中央和高架十字迷宫开放臂停留的时间和总路程减少（<0.05）；模型组BNST组织神经元数量减少，细胞排列紊乱，细胞萎缩，核固缩，神经元结构异常，尼氏染色不均匀，尼氏体减少。模型组p-PKC蛋白表达及p-PKC/PKC比值升高（<0.05），p-ERK1/2、p-CREB蛋白表达及p-ERK1/2/ERK1/2比值降低（<0.05）。与模型组比较，药物组和针刺组大鼠在旷场中央和高架十字迷宫开放臂停留的时间和总路程增加（<0.05）；BNST组织神经元数量增加，细胞排列更有序，神经元结构改善，尼氏体增多；药物组和针刺组p-PKC蛋白表达及p-PKC/PKC比值降低（<0.05），p-ERK1/2、p-CREB蛋白表达及p-ERK1/2/ERK1/2比值升高（<0.05）。
针刺可缓解PTSD大鼠的焦虑恐惧行为，改善BNST神经元损伤。其机制可能与调节BNST中PKC/ERK/CREB信号通路有关。
