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甘肃中医药大学针灸推拿学院,兰州 730000
王永蕊,甘肃中医药大学硕士研究生。E-mail:910273938@qq.com
✉严兴科,教授。E-mail:yanxingke@126.com
收稿:2023-11-16,
网络首发:2024-09-02,
纸质出版:2024-11-12
移动端阅览
王永蕊, 郑先丽, 严兴科. “疏肝调神”法针刺对创伤后应激障碍大鼠焦虑样行为及终纹床核PKC/ERK/CREB通路的影响[J]. 中国针灸, 2024,44(11):1281-1288.
WANG Yongrui, ZHENG Xianli, YAN Xingke. Effects of
王永蕊, 郑先丽, 严兴科. “疏肝调神”法针刺对创伤后应激障碍大鼠焦虑样行为及终纹床核PKC/ERK/CREB通路的影响[J]. 中国针灸, 2024,44(11):1281-1288. DOI: 10.13703/j.0255-2930.20231116-k0006.
WANG Yongrui, ZHENG Xianli, YAN Xingke. Effects of
目的:
2
观察“疏肝调神”法针刺对创伤后应激障碍(PTSD)大鼠终纹床核(BNST)蛋白激酶C/细胞外信号调节蛋白激酶/cAMP反应元件结合蛋白(PKC/ERK/CREB)信号通路的影响,探讨针刺改善PTSD焦虑与恐惧的作用机制。
方法:
2
从50只SPF级雄性SD大鼠中随机选出10只作为空白组,剩余40只 大鼠采用幽闭电击法联合负重力竭式游泳法制备PTSD大鼠模型。随机选取30只模型制备成功的大鼠,分为模型组、西药组与针刺组,各10只。西药组予盐酸帕罗西汀溶液灌胃,每日1次,连续12 d;针刺组针刺“百会”及双侧“内关”“神门”“太冲”,“百会”每日均针刺,其余穴位单日针刺左侧,双日针刺右侧,每日1次,连续12 d。采用旷场实验和高架十字迷宫实验检测大鼠焦虑与恐惧的行为学变化;利用HE染色及尼氏染色观察大鼠BNST组织形态;Western blot法检测大鼠BNST组织PKC、磷酸化蛋白激酶C(p-PKC)、细胞外信号调节蛋白激酶1/2(ERK1/2)、磷酸化细胞外信号调节蛋白激酶1/2(p-ERK1/2)、磷酸化CAMP反应元件结合蛋白(p-CREB)蛋白表达。
结果:
2
与空白组比较,模型组大鼠在旷场中央区域停留时间及停留总距离缩短、在高架十字迷宫开放臂停留时间及活动总距离缩短(
P
<
0.05);BNST组织神经细胞数量减少,排列不规律,存在细胞皱缩及核固缩,神经元结构异常,尼氏体染色不均匀,数量减少;BNST组织p-PKC蛋白表达及p-PKC/PKC值升高(
P
<
0.05),p-ERK1/2、p-CREB蛋白表达及p-ERK1/2/ERK1/2值降低(
P
<
0.05)。与模型组比较,西药组、针刺组大鼠在旷场中央区域停留时间及停留总距离增加、在高架十字迷宫开放臂停留时间及活动总距离增加(
P
<
0.05);BNST组织神经细胞数量较模型组增多,细胞排列尚整齐,尼氏体数量增加,神经元结构、尼氏体形态改善;BNST组织p-PKC蛋白表达及p-PKC/PKC值降低(
P
<
0.05),p-ERK1/2、p-CREB蛋白表达及p-ERK1/2/ERK1/2值升高(
P
<
0.05)。
结论:
2
“疏肝调神”法针刺可减轻PTSD大鼠的焦虑与恐惧样行为,改善BNST组织神经损伤,其机制可能与调节BNST组织PKC/ERK/CREB信号通路有关。
Objective
2
To observe the effect of
Shugan Tiaoshen
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.
Methods
2
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.
Results
2
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 (
P
<
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 (
P
<
0.05) and decreased protein expression of p-ERK1/2
p-CREB
and p-ERK1/2/ERK1/2 ratio (
P
<
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 (
P
<
0.05); the BNST tissues showed increased number of neurons
mor
e 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 (
P
<
0.05) and increased p-ERK1/2
p-CREB protein expression
and p-ERK1/2/ERK1/2 ratio (
P
<
0.05).
Conclusion
2
Shugan Tiaoshen
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.
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