Effect of <italic style="font-style: italic">ZHU Lian</italic>ʹs acupuncture stimulation method on neurological function in rats with ischemic stroke based on the miR-34a-5p/YWHAG pathway

ZHAI Yang; LI Fang; WANG Han; MO Zhizhen; WEI Yulu; CHI Tengfei; ZOU Min; PANG Yan; MEI Xiaoping

doi:10.13703/j.0255-2930.20241128-0001

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Effect of ZHU Lianʹs acupuncture stimulation method on neurological function in rats with ischemic stroke based on the miR-34a-5p/YWHAG pathway

Study on Mechanism | 更新时间：2026-03-16

- Effect of ZHU Lianʹs acupuncture stimulation method on neurological function in rats with ischemic stroke based on the miR-34a-5p/YWHAG pathway
- Chinese Acupuncture & Moxibustion Vol. 46, Issue 3, Pages: 375-383(2026)
- 作者机构：
  
  南宁市第七人民医院/南宁市针灸研究所脑病科，广西南宁　530012
  南宁市第七人民医院/南宁市针灸研究所针灸康复一区，广西南宁　530012
  广西国际壮医医院/广西中医药大学附属国际壮医医院儿科，南宁　530200
  广西中医药大学第一附属医院急诊科
  广西国际壮医医院/广西中医药大学附属国际壮医医院脑病科，南宁　530200
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20241128-0001
  CLC：
- Received：28 November 2024，
  
  Online First：04 December 2025，
  
  Published：12 March 2026
- 稿件说明：
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ZHAI Yang, LI Fang, WANG Han, et al. Effect of ZHU Lianʹs acupuncture stimulation method on neurological function in rats with ischemic stroke based on the miR-34a-5p/YWHAG pathway[J]. Chinese Acupuncture & Moxibustion, 2026, 46(3): 375-383. DOI: 10.13703/j.0255-2930.20241128-0001.
DOI：

ZHAI Yang, LI Fang, WANG Han, et al. Effect of ZHU Lianʹs acupuncture stimulation method on neurological function in rats with ischemic stroke based on the miR-34a-5p/YWHAG pathway[J]. Chinese Acupuncture & Moxibustion, 2026, 46(3): 375-383. DOI: 10.13703/j.0255-2930.20241128-0001. DOI：

摘要

目的：

观察朱琏针刺兴奋法对缺血性脑卒中（IS）模型大鼠神经功能及微小RNA-34a-5p（miR-34a-5p）/酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白γ（YWHAG）通路的影响，探究其减轻IS神经功能损伤的作用机制。

方法：

在72只清洁级雄性SD大鼠中随机选取10只为假手术组，其余采用线栓法建立IS模型，将造模成功的50只大鼠随机分为模型组、针刺组、阳性药物组、针刺+激动剂对照组和针刺+激动剂组，每组10只。针刺组穴取“水沟”“百会”“大椎”及双侧“内关”“合谷”等，以朱琏针刺兴奋法进行操作；阳性药物组以10.8 mg/kg尼莫地平溶液灌胃；针刺+激动剂组、针刺+激动剂对照组分别进行脑室内注射miR-34a-5p激动剂（miR-34a-5p agomir）及阴性对照剂（agomir NC），注射剂量为5μL，随后按照针刺组进行操作。针刺及用药均每天1次，干预5 d后间隔2 d，连续21 d。干预后，采用Zea-Longa评分评估大鼠神经功能受损程度；TTC染色法检测大鼠脑梗死体积百分比；尼氏染色观察大鼠缺血侧皮质组织神经元损伤及尼氏小体情况；TUNEL染色检测大鼠缺血侧皮质组织神经元凋亡率；实时荧光定量PCR法检测大鼠缺血侧皮质组织miR-34a-5p、YWHAG mRNA表达；Western blot法检测大鼠缺血侧皮质组织白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和YWHAG蛋白表达；双荧光素酶实验验证miR-34a-5p、YWHAG关系。

结果：

与假手术组比较，模型组大鼠Zea-Longa评分、脑梗死体积百分比、缺血侧皮质组织神经元凋亡率、IL-6、TNF-α及miR-34a-5p mRNA表达升高（

0.05），YWHAG mRNA及蛋白表达降低（

0.05）。与模型组比较，针刺组、阳性药物组、针刺+激动剂对照组大鼠Zea-Longa评分、脑梗死体积百分比、缺血侧皮质组织神经元凋亡率、IL-6、TNF-α及miR-34a-5p mRNA表达降低（

0.05），YWHAG mRNA及蛋白表达升高（

0.05）；与针刺+激动剂对照组比较，针刺+激动剂组大鼠Zea-Longa评分、脑梗死体积百分比、缺血侧皮质组织神经元凋亡率、IL-6、TNF-α及miR-34a-5p mRNA表达升高（

0.05），YWHAG mRNA及蛋白表达降低（

0.05）。miR-34a-5p、YWHAG具有靶向调节关系（

0.05）。

结论：

朱琏针刺兴奋法可改善IS模型大鼠神经功能，其机制可能与调控miR-34a-5p/YWHAG通路有关。

Abstract

Objective

To observe the effects of the

ZHU Lian

's acupuncture stimulation method on neurological function and miR-34a-5p/tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein γ (YWHAG) pathway in rats with ischemic stroke (IS)

and to explore its mechanism in alleviating neurological impairment in IS.

Methods

A total of 72 clean-grade male SD rats

with 10 rats were randomly selected as the sham operation group

and the remaining rats were subjected to suture occlusion method to establish an IS model. Fifty successfully modeled rats were randomly divided into a model group

an acupuncture group

a positive drug group

an acupuncture+agonist control group

and an acupuncture+agonist group

with 10 rats in each group. The rats in the acupuncture group were treated with

ZHU Lian

's acupuncture stimulation method at "Shuigou" (GV26)

"Baihui" (GV20)

"Dazhui" (GV14)

and bilateral "Neiguan" (PC6) and "Hegu" (LI4). The rats in the positive drug group were treated with 10.8 mg/kg nimodipine via gavage. The rats in the acupuncture+agonist group and acupuncture+agonist control group were treated with intracerebroventri

cular injections of miR-34a-5p agonist (miR-34a-5p agomir) and negative control (agomir NC)

respectively

at a dose of 5μL

followed by the same acupuncture protocol. All treatments were performed once daily for 5 days

followed by 2 days off

continuing for 21 days. After intervention

neurological deficits were evaluated using the Zea-Longa score. The infarct volume percentage was measured by TTC staining. Neuronal damage and Nissl bodies in the ischemic cortex were observed by Nissl staining. Neuronal apoptosis rate in the ischemic cortex was assessed by TUNEL staining. Relative expression levels of miR-34a-5p and YWHAG mRNA in the ischemic cortex were measured by RT-qPCR. Protein levels of interleukin-6 (IL-6)

tumor necrosis factor-α (TNF-α)

and YWHAG were detected by Western blot. A dual-luciferase assay was used to verify the relationship between miR-34a-5p and YWHAG.

Results

Compared with the sham operation group

the Zea-Longa scores

the percentage of cerebral infarct volume

the neuronal apoptosis rate in the ischemic cortical tissue

the expression levels of IL-6

TNF-α

and miR-34a-5p mRNA in the model group were increased (

0.05)

and the expression of YWHAG mRNA and protein was decreased (

0.05). Compared with the model group

the Zea-Longa scores

the percentage of cerebral infarct volume

the neuronal apoptosis rate in the ischemic cortical tissue

the expression levels of IL-6

TNF-α

and miR-34a-5p mRNA in the acupuncture group

the positive drug group

and the acupuncture+agonist control group were decreased (

0.05)

and the expression of YWHAG mRNA and protein was increased (

0.05). Compared with the acupuncture+agonist control group

the Zea-Longa scores

the percentage of cerebral infarct volume

the neuronal apoptosis rate in the ischemic cortical tissue

the expression levels of IL-6

TNF-α

and miR-34a-5p mRNA in the acupuncture+ agonist group were increased (

0.05)

and the expression of YWHAG mRNA and protein was decreased (

0.05). In addition

the miR-34a-5p and YWHAG have a targeted regulatory relationship (

0.05).

Conclusion

The

ZHU Lian

ʹs acupuncture stimulation method could improve neurological function in IS model rats

possibly by modulating the miR-34a-5p/YWHAG pathway.

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⁰

Effect of ZHU Lianʹs acupuncture stimulation method on neurological function in rats with ischemic stroke based on the miR-34a-5p/YWHAG pathway

DOI：10.13703/j.0255-2930.20241128-0001

摘要

Abstract

关键词

Keywords

references