头针疗法联合重复经颅磁刺激对脑卒中大鼠神经功能的改善作用及对 PKA/ CREB 信号通路的影响
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1.河北北方学院附属第一医院康复医学科,河北 张家口 075000; 2.河北北方学院附属第一医院神经内科,河北 张家口 075000

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R-33

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Effect of scalp acupuncture combined with repeated transcranial magnetic stimulation on neurological function in rats with stroke and its effect on the PKA / CREB signaling pathway
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1.Department of Rehabilitation Medicine, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China. 2. Department of Neurology, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000

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    摘要:

    目的 探讨头针疗法联合重复经颅磁刺激( rTMS)对脑卒中大鼠神经功能的改善作用及对蛋白激 酶 A(PKA) / 环磷酸腺苷反应元件结合蛋白(CREB)信号通路的影响。 方法 50 只大鼠随机分为假手术组、模型组、头针组、rTMS 组、联合组,各 10 只;假手术组除不插入尼龙线栓,其余各组应用线栓法制备局灶性脑卒中大鼠模型。 模型组 9 只、头针组 8 只、rTMS 组 8 只、联合组 9 只建模成功,模型组和假手术组不做干预,头针组给予头针疗 法,rTMS 组给予 rTMS,联合组给予头针疗法+rTMS,末次干预后进行神经功能缺损程度评分、Morris 水迷宫实验,采用 TTC 及 HE 染色观察脑组织梗死灶情况及梗死灶周围皮质病理学改变,并测定脑组织 PKA、CREB 蛋白及磷酸化 CREB(p-CREB)蛋白表达量。 结果 与假手术组比较,模型组、头针组、rTMS 组及联合组神经功能损伤评分升高, 逃避潜伏期均延长,目标象限停留时间均缩短(P<0. 05);与模型组比较,头针组、rTMS 组及联合组神经功能损伤评 分降低,逃避潜伏期均缩短,目标象限停留时间均延长(P<0. 05);与头针组和 rTMS 组比较,联合组神经功能损伤 评分降低,逃避潜伏期缩短,目标象限停留时间延长(P<0. 05)。 TTC 染色结果显示,模型组缺血侧大脑皮层可见 白色梗死灶,并伴有轻度糜烂;头针组、rTMS 组和联合组缺血侧大脑皮层梗死面积缩小,脑组织外观有所改善,联 合组改善效果最为明显;HE 染色结果显示,模型组脑组织结构疏松,呈现筛网状液化性坏死状态,神经元大量消失,多量小胶质细胞浸润;头针组和 rTMS 组脑组织结构破坏轻于模型组,仍可见部分正常神经元形态,但部分细胞质呈现空泡化、水肿状态;联合组脑组织结构及神经元形态趋于正常。 与假手术组比较,模型组、头针组、rTMS 组 及联合组 PKA 蛋白及 p-CREB 蛋白相对表达量均降低(P<0. 05);与模型组比较,头针组、rTMS 组及联合组 PKA、p- CREB 蛋白相对表达量均升高(P<0. 05),且联合组高于头针组和 rTMS 组(P<0. 05)。 结论 头针疗法联合 rTMS 可有效改善脑卒中大鼠神经功能,其作用机制可能通过激活 PKA/ CREB 信号通路发挥调控作用。

    Abstract:

    Objective To investigate the effect of scalp acupuncture combined with repeated transcranial magnetic stimulation (rTMS) on neurological function in stroke rats and its effect on the protein kinase A (PKA) / cyclophosphine response element binding ( CREB) protein signaling pathway. Methods Fifty rats were randomly divided into a sham operation group, model group, scalp group, rTMS group, and combined group (10 rats per group). Nylon thread was not inserted in the sham operation group. The other groups were used to prepare a model of focal stroke rats using sutures. Nine rats in the model group, eight in the scalp group, eight in the rTMS group, and nine in the combined group were successfully modeled. Rats in the model group and sham operation group did not receive the intervention. The scalp group received scalp acupuncture, the rTMS group received rTMS, and the combined group received scalp acupuncture + rTMS. After the last intervention, neurological deficit score and Morris water maze test performance were assessed. After triphenyl tetrazolium chloride (TTC) and hematoxylin and eosin (HE) staining, the pathological cortical changes in and around the infarct of brain tissues were observed. The expressions of PKA, CREB protein, and phosphorylated CREB ( p-CREB) protein were measured. Results Compared with the sham operation group, the model group, scalp group, rTMS group, and combined group had higher neurological injury scores, longer escape latency, and shorter target quadrant stay duration (P< 0. 05). Compared with the model group, the scalp group, rTMS group, and combination group showed lower neurological injury scores, shorter escape latency, and longer target quadrant stay duration (P<0. 05). Compared with the scalp group and the rTMS group, the combined group showed decreased neurological injury score, shortened escape latency, and prolonged target quadrant stay duration (P<0. 05). TTC staining identified white infarcts in the cerebral cortex of the ischemic side of the model group and mild erosion. The infarct area of the cerebral cortex of the ischemic side of the scalp acupuncture group, rTMS group, and combination group reduced and the appearance of the brain tissue improved; the improvement effect was most obvious in the combination group. The brain tissue structure of the model group was loose and showed liquefying necrosis in the cribriform network, with a large number of neurons disappearing and a large number of microglia infiltrating. The brain tissue structure of the scalp acupuncture group and rTMS group showed less damage than that of the model group, with some remaining normal neurons. However, some cytoplasm showed vacuolation and edema. The brain tissue structure and neuron morphology of the combined group tended to be positive. Compared with the sham operation group, the model group, scalp acupuncture group, rTMS group, and combination group showed decreased relative expressions of PKA and p-CREB protein (P<0. 05). Compared with the model group, the scalp acupuncture group, rTMS group, and combination group showed increased relative expressions of PKA and p-CREB protein ( P < 0. 05 ); the expression was higher in the combined group than in the scalp acupuncture group and rTMS group (P<0. 05). Conclusions Scalp acupuncture combined with rTMS can effectively improve neurological function in stroke rats. Its mechanism may play a regulatory role by activating the PKA/ CREB signaling pathway.

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戈 蕾,邹玉安,王晓娜.头针疗法联合重复经颅磁刺激对脑卒中大鼠神经功能的改善作用及对 PKA/ CREB 信号通路的影响[J].中国比较医学杂志,2020,30(10):85~91.

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  • 收稿日期:2020-03-07
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  • 在线发布日期: 2020-11-25
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