PARP9通过抑制PI3K/AKT/mTOR信号通路促进脑缺血再灌注大鼠神经元凋亡及炎症反应而加重认知障碍

陈丽; 张德绸; 叶丽莎

doi:10.16016/j.2097-0927.202605059

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PARP9通过抑制PI3K/AKT/mTOR信号通路促进脑缺血再灌注大鼠神经元凋亡及炎症反应而加重认知障碍

神经科学 | 更新时间：2026-10-19

- PARP9通过抑制PI3K/AKT/mTOR信号通路促进脑缺血再灌注大鼠神经元凋亡及炎症反应而加重认知障碍
- PARP9 promotes neuronal apoptosis and inflammatory responses through inhibiting the PI3K/AKT/mTOR signaling pathway and then aggravates cognitive impairment in rats with cerebral ischemia-reperfusion
- 陆军军医大学学报 2026年48卷第19期页码：2770-2779
- 作者机构：
  
  西南医科大学附属中医医院神经内科，四川泸州
- 作者简介：
  
  叶丽莎，E-mail：1873936902@qq.com
- 基金信息：
  
  四川省自然科学基金面上项目(2024NSFSC1823);西南医科大学中西医结合特色项目(2023ZYYQ02);四川省科技计划项目(2022YFS0613)
- DOI：10.16016/j.2097-0927.202605059
  中图分类号： R363.21;R338.64;R743.31
- 收稿：2026-05-21，
  
  修回：2026-08-09，
  
  录用：2026-08-19，
  
  纸质出版：2026-10-15
- 稿件说明：
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陈丽，张德绸，叶丽莎.PARP9通过抑制PI3K/AKT/mTOR信号通路促进脑缺血再灌注大鼠神经元凋亡及炎症反应而加重认知障碍［J］.陆军军医大学学报，2026，48（19）：2770-2779.DOI：10.16016/j.2097-0927.202605059.

Chen L，Zhang D C，Ye L S.PARP9 promotes neuronal apoptosis and inflammatory responses through inhibiting the PI3K/AKT/mTOR signaling pathway and then aggravates cognitive impairment in rats with cerebral ischemia-reperfusion［J］.J Army Med Univ，2026，48（19）：2770-2779.DOI：10.16016/j.2097-0927.202605059.
陈丽，张德绸，叶丽莎.PARP9通过抑制PI3K/AKT/mTOR信号通路促进脑缺血再灌注大鼠神经元凋亡及炎症反应而加重认知障碍［J］.陆军军医大学学报，2026，48（19）：2770-2779.DOI：10.16016/j.2097-0927.202605059. DOI：

Chen L，Zhang D C，Ye L S.PARP9 promotes neuronal apoptosis and inflammatory responses through inhibiting the PI3K/AKT/mTOR signaling pathway and then aggravates cognitive impairment in rats with cerebral ischemia-reperfusion［J］.J Army Med Univ，2026，48（19）：2770-2779.DOI：10.16016/j.2097-0927.202605059. DOI：

摘要

目的

脑缺血再灌注（cerebral ischemia-reperfusion， CIR）是指血流恢复后加重缺血区神经元损伤的病理过程，其机制涉及氧化应激、炎症及凋亡等级联反应。聚（ADP-核糖）聚合酶9 ［poly（ADP-ribose）polymerase 9， PARP9］在CIR中的生物学功能尚待阐明。本研究旨在探讨PARP9对CIR大鼠神经元凋亡、神经炎症、认知能力及PI3K/AKT/mTOR信号通路的影响。

方法

①损伤效应实验：10只SD大鼠［雄性，8~10周龄，体质量250~260 g］采用随机数字表法分为CIR组和假手术组，每组5只。CIR组大鼠采用线栓法实现大脑中动脉栓塞，2 h后拔线再通，建立CIR模型；假手术组大鼠只暴露血管，不进行切口和插入石蜡线栓。术后7 d，Western blotting法检测2组大鼠脑组织中PARP9蛋白表达水平。②PARP9干预实验：60只SD大鼠［雄性，8~10周龄，体质量250~260 g］采用随机数字表法分为AAV-siScramble组、AAV-siPARP9组、AAV-Scramble组、AAV-PARP9组，每组15只。各组大鼠脑室注射相应腺病毒（2 μL），于注射相应腺病毒2周后，各组大鼠构建CIR模型。术后24 h，Zea-Longa法评估各组大鼠神经功能；Morris水迷宫评估各组大鼠认知能力。术后7 d，TTC染色检测各组大鼠脑梗死面积；Nissl染色检测各组大鼠脑皮质病理损伤情况；TUNEL染色检测各组大鼠脑组织中神经元凋亡率；免疫组织化学染色检测各组大鼠脑组织中TNF-α、IL-1β阳性表达；Western blotting法检测各组大鼠脑组织中PARP9、p-PI3K、p-AKT、p-mTOR蛋白表达。

结果

①损伤效应实验：CIR组大鼠脑组织缺血皮层区中PARP9的表达水平较假手术组显著增加（

0.001）。②PARP9干预实验：与AAV-siScramble组比较，AAV-siPARP9组大鼠的神经功能评分降低，逃避潜伏期缩短，原平台象限停留次数增加（

0.001）；大鼠脑梗死面积占比明显下降（

0.05）；脑组织缺血皮层区中PARP9表达水平、神经元凋亡率、TNF-α和IL-1β的阳性表达均下降（

0.05）；脑组织缺血皮层区中p-PI3K、p-AKT、p-mTOR蛋白表达增加（

0.05）；与AAV-Scramble组比较，AAV-PARP9组大鼠的神经功能评分增加，逃避潜伏期增加，原平台象限停留次数减少（

0.001）；脑梗死面积占比显著增加（

0.05）；脑组织缺血皮层区中PARP9表达水平、神经元凋亡率、TNF-α和IL-1β的阳性表达均增加（

0.05）；脑组织缺血皮层区中p-PI3K、p-AKT、p-mTOR蛋白表达下降（

0.05）。AAV-siPARP9组大鼠大脑皮质神经元排列规整，尼氏体含量丰富，胞质呈蓝色，细胞核着色均匀；AAV-PARP9组则表现为皮质神经元分布散乱，局部可见神经元数量减少，胞质蓝染程度变浅，尼氏体明显减少；而AAV-siScramble组和AAV-Scramble组神经元尼氏体溶解现象有所缓解，神经元存活数量增加。

结论

PARP9是CIR病理损伤的重要促进因子，其可能通过抑制PI3K/AKT/mTOR信号通路介导。靶向抑制PARP9可减轻神经元凋亡和神经炎症反应，改善认知功能，提示PARP9可能为CIR干预提供新的分子靶点。

Abstract

Objective

Cerebral ischemia-reperfusion (CIR) refers to a pathological process in which restoration of blood flow exacerbates neuronal injury in the ischemic area

with the underlying mechanisms involving cascade reactions such as oxidative stress

inflammation

and apoptosis. The biological function of poly(ADP-ribose) polymerase 9 (PARP9) in CIR remains unclarified. This study aims to investigate the effects of PARP9 on neuronal apoptosis

neuroinflammation

cognitive function

and the PI3K/AKT/mTOR signaling pathway in rats with CIR.

Methods

① Experiments of injury effects: Ten male SD rats (8 to 10 weeks old

weighing 250 to 260 g) were randomly divided into a sham operation group and a CIR group

with 5 rats in each group. Rats in the CIR group underwent middle cerebral artery occlusion using intraluminal suture technique

followed by reperfusion 2 h later by removing the suture to establish the CIR model; rats in the sham operation group only had the blood vessels exposed without incision or insertion of a paraffin-coated suture. At 7 d after surgery

Western blotting was applied to detect the protein levels PARP9 in the brain tissues of both groups. ② PARP9 intervention experiment: 60 male SD rats (8 to 10 weeks old

weighing 250 to 260 g) were randomly divided into an AAV-siScramble group

an AAV-siPARP9 group

an AAV-Scramble group

and an AAV-PARP9 group

with 15 rats in each group. Rats in each group received an intracerebroventricular injection of the corresponding adenovirus (3 μL)

and the CIR model was established 2 weeks after adenovirus injection. At 24 h postoperatively

neurological function was assessed using Zea-Longa method

and cognitive function was evaluated using Morris water maze test. At 7 d after surgery

TTC staining and Nissl staining were used to measure cerebral infarct volume and evaluate pathological damage in the cerebral cortex

respectively. TUNEL staining was used to detect the neuronal apoptosis rate; immunohistochemical staining was employed to detect the positive expression of TNF-α and IL-1β; and Western blotting was utilized to detect the protein expression of PARP9

p-PI3K

p-AKT

and p-mTOR in brain tissue.

Results

① Experiments of injury effects: The expression level of PARP9 in the ischemic cortical area of brain tissue was significantly increased in the CIR group than the sham operation group (

0.001). ② PARP9 intervention experiment: Compared with the AAV-siScramble group

the AAV-siPARP9 group exhibited decreased neurological function scores

shortened escape latency

and increased number of platform quadrant crossings (

0.001). The percentage of cerebral infarct area was significantly reduced (

0.05). The expression level of PARP9

neuronal apoptosis rate

and positive expression levels of TNF-α and IL-1β in the ischemic cortical area were all decreased (

0.05). The protein levels of p-PI3K

p-AKT

and p-mTOR in the ischemic cortical area were increased (

0.05). Compared with the AAV-Scramble group

the AAV-PARP9 group showed increased neurological function scores

prolonged escape latency

and reduced number of platform quadrant crossings (

0.001). The percentage of cerebral infarct area was significantly increased (

0.05). The expression level of PARP9

neuronal apoptosis rate

and positive expression of TNF-α and IL-1β in the ischemic cortical area were all increased (

0.05). The protein levels of p-PI3K

p-AKT

and p-mTOR in the ischemic cortical area were decreased (

0.05). In the AAV-siPARP9 group

cortical neurons were regularly arranged

with abundant Nissl bodies

blue-stained cytoplasm

and uniformly stained nuclei. In the AAV-PARP9 group

cortical neurons were scattered with locally reduced neuronal numbers

lighter blue-stained cytopla

and significantly reduced Nissl bodies; while in the AAV-siScramble group and AAV-Scramble group

the dissolution of Nissl bodies was alleviated and the number of surviving neurons was increased.

Conclusion

PARP9 functions as a key pathogenic facilitator in CIR injury

which may be mediated through inhibition of the PI3K/AKT/mTOR signaling cascade. Knockdown of PARP9 alleviates neuronal apoptosis and neuroinflammatory responses

and improve cognitive function

suggesting that PARP9 may serve as a new molecular target for CIR intervention.

关键词

Keywords

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