头顶一颗珠通过调节内质网自噬保护卒中后认知障碍大鼠的神经功能

doi:10.12122/j.issn.1673-4254.2026.06.016

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (6): 1365-1373.doi: 10.12122/j.issn.1673-4254.2026.06.016

• • 上一篇

头顶一颗珠通过调节内质网自噬保护卒中后认知障碍大鼠的神经功能

杨丹¹^,³(), 赵方毓¹^,²^,³, 何一多¹^,², 朱红¹^,², 刘昕¹^,², 陈显兵¹^,²^,³()

^1.风湿性疾病发生与干预湖北省重点实验室，湖北恩施 445000
^2.湖北民族大学附属民大医院病理研究所，湖北恩施 445000
^3.湖北民族大学医学部，湖北恩施 445000

收稿日期:2025-12-16 出版日期:2026-06-20 发布日期:2026-06-24
通讯作者: 陈显兵 E-mail:820214831@qq.com;chenxianbing7612@163.com
作者简介:杨丹，硕士研究生，医师，E-mail: 820214831@qq.com
基金资助:
国家自然科学基金(82260821);研究生教育创新项目(MYK2023072);湖北民族大学科研项目(XN2303);风湿性疾病发生与干预湖北省重点实验室青年项目(OIR202407Q)

Trillium tschonoskii Maxim saponin protects neurological function in rats with post-stroke cognitive impairment by promoting ER-phagy

Dan YANG¹^,³(), Fangyu ZHAO¹^,²^,³, Yiduo HE¹^,², Hong ZHU¹^,², Xin LIU¹^,², Xianbing CHEN¹^,²^,³()

^1.Hubei Provincial Key Laboratory of Occurrence and Intervention of Rhumatic Diseases, Enshi 445000, China
^2.Pathology Institute of Minda Hospital Affiliated to Hubei Minzu University, Enshi 445000, China
^3.Medical School, Hubei Minzu University, Enshi 445000, China

Received:2025-12-16 Online:2026-06-20 Published:2026-06-24
Contact: Xianbing CHEN E-mail:820214831@qq.com;chenxianbing7612@163.com
Supported by:
National Natural Science Foundation of China(82260821)

摘要/Abstract

摘要：

目的探讨头顶一颗珠（TTM）通过调节内质网自噬对卒中后认知障碍（PSCI）大鼠的神经保护作用。方法通过改良线栓法建立卒中后认知障碍大鼠模型，分为Sham组、Model组、TTM组、自噬诱导剂雷帕霉素组（Rap）、自噬抑制剂3-甲基-3H-嘌呤-6-胺组（3-MA）及TTM+3-MA组，10只/组，对应处理各组大鼠。采用HE染色、Nissl染色观察各组大鼠脑组织病理学变化，TUNEL染色观察各组大鼠神经元凋亡情况，Western blotting法检查各组大鼠内质网自噬相关蛋白及凋亡蛋白的表达。采用Morris水迷宫观察各组大鼠学习记忆功能，采用免疫组织化学法观察自噬相关5（ATG5）与葡萄糖调节蛋白78 （GRP78）的阳性表达；采用高尔基染色观察神经元树突棘数量；采用免疫荧光观察各组大鼠脑组织中NeuN的表达及网状吞噬调节剂 1（FAM134B）、钙连蛋白Calnexin与微管相关蛋白轻链 3（LC3）的共定位表达；采用Western blotting检测各组大鼠海马组织中FAM134B、ATG5、鳌合体1（P62）、LC3、GRP78、Bcl-2相关X蛋白（Bax）、B淋巴细胞瘤-2基因（BCL-2）、白介素-10 （IL-10）、白细胞介素-1β （IL-1β）、肿瘤坏死因子-α（TNF-α）蛋白的表达。结果与Sham组对比，Model组大鼠逃避潜伏的过程中时间明显增加，在目标区域停留时间则有所缩短，穿越该平台次数有所减少（P<0.05）；组织病理学示大鼠海马组织细胞数量减少，排列稀疏、紊乱，核固缩，空泡化严重；尼氏小体数量减少；脑组织中ATG5与GRP78阳性表达增加；神经元凋亡增加（P<0.05），FAM134B与LC3、Calnexin与LC3共定位稍增加。Western blotting结果显示海马组织中IL-1β、TNF-α、Bax蛋白表达升高（P<0.05），IL-10、BCL-2蛋白表达下降（P<0.05），GRP78、FAM134B、P62蛋白表达上调（P<0.05），LC3表达下调。与Model组对比，TTM组大鼠逃避潜伏时间有所缩短，在目标象限停留时间增加，穿越平台次数增多（P<0.05）；组织病理学示ATG5表达进一步增加，而GRP78表达减少；神经元树突棘增加，FAM134B与LC3、Calnexin与LC3共定位增加。Western blotting结果显示IL-1β、TNF-α、Bax、GRP78及P62蛋白表达下降（P<0.05），FAM134B、ATG5、LC3、IL-10和BCL-2蛋白表达升高（P<0.05）；3-MA组与TTM组趋势相反。结论卒中早期细胞内质网应激被过度激活，适应性信号转变为促凋亡信号，内质网自噬虽被激活但通量不足，无法减轻细胞组织损伤、神经元凋亡，导致神经功能下降伤。TTM可通过调节内质网自噬，缓解过度激活的内质网应激，减轻大鼠脑组织病理损伤，减轻炎症反应与凋亡，减少神经元树突棘丢失，从而对卒中后认知障碍大鼠认知功能起保护作用。

关键词: 头顶一颗珠, 内质网自噬, 卒中后认知障碍, 神经保护

Abstract:

Objective To investigate the mechanism that mediates the neuroprotective effects of Trillium tschonoskii Maxim (TTM) against post-stroke cognitive impairment (PSCI) in rats. Methods Adult SD rats were randomized into Sham operation, PSCI model, TTM, rapamycin (an autophagy inducer), 3-methyladenine (an autophagy inhibitor), and TTM+3-MA groups, and rat models of cognitive impairment were established using a modified thread occlusion method. Cognitive function of the rats was assessed using Morris water maze test. Histopathological changes, neuronal apoptosis, dendritic spines, and protein expressions of FAM134B, LC3, ATG5, P62, GRP78, Bax, Bcl-2, IL-10, IL-1β, and TNF-α were evaluated using HE, Nissl, TUNEL, Golgi staining, immunohistochemistry, immunofluorescence staining, and Western blotting. Results Compared with the sham-operated rats, the rat models of PSCI showed significantly prolonged escape latency, reduced target quadrant time and platform crossings, severe hippocampal damage, increased ATG5 and GRP78 expression, elevated apoptosis, increased IL-1β, TNF-α, Bax, GRP78, and P62 expressions, and decreased IL-10, Bcl-2, and LC3 expressions, with slightly increased FAM134B-LC3 and calnexin-LC3 co-localization. Compared with those in the model group, the rats receiving TTM treatment showed significantly shortened escape latency, increased target quadrant time and platform crossings, increased ATG5 and dendritic spines, decreased GRP78 expression, enhanced FAM134B-LC3 and calnexin-LC3 co-localization, reduced IL-1β, TNF-α, Bax, GRP78, and P62 expressions, and increased FAM134B, ATG5, LC3, IL-10, and Bcl-2 expressions; the rats treated with 3-MA showed the opposite changes. Conclusion Excessive ER stress is activated early after stroke, shifting from adaptive to pro-apoptotic signaling, with insufficient ER-phagy flux. TTM modulates ER-phagy, alleviates ERS, reduces neuroinflammation and apoptosis, protects dendritic spines, and improves cognitive function in rats with PSCI.

Key words: Trillium tschonoskii Maxim saponin, ER-phagy, post-stroke cognitive impairment, neuroprotection

杨丹, 赵方毓, 何一多, 朱红, 刘昕, 陈显兵. 头顶一颗珠通过调节内质网自噬保护卒中后认知障碍大鼠的神经功能[J]. 南方医科大学学报, 2026, 46(6): 1365-1373.

Dan YANG, Fangyu ZHAO, Yiduo HE, Hong ZHU, Xin LIU, Xianbing CHEN. Trillium tschonoskii Maxim saponin protects neurological function in rats with post-stroke cognitive impairment by promoting ER-phagy[J]. Journal of Southern Medical University, 2026, 46(6): 1365-1373.

图/表 7

图1 内质网自噬可以减轻卒中后认知障碍大鼠脑组织病理损伤

Fig.1 ER-phagy alleviates histopathological damage in the brain of rats with post-stroke cognitive impairment (PSCI). A: HE staining and Nissl staining of rat hippocampal tissues in each group (Original magnification: ×40). B: TUNEL staining for observing neuronal apoptosis in each group (Scale bar=50 μm). *P<0.05, #P<0.05, ##P<0.01 vs Model group (n=3).

图2 FAM134B, GRP78, P62, Bax, Bcl-2和LC3蛋白表达情况

Fig.2 Western blotting for detecting expression levels of FAM134B, GRP78, P62, Bax, Bcl-2, and LC3 proteins in the hippocampal tissue of rats in each group (n=4). *P<0.05, **P<0.01 vs Sham group; #P<0.05, ##P<0.01 vs Model group; &&P<0.01 vs 3-MA group.

图3 各组大鼠水迷宫实验结果

Fig.3 Results of the Morris water maze test in each group (n=10). A: Escape latency over 5 days in each group. B: Time spent in the target quadrant. C: Number of platform crossings. *P<0.05 vs Sham group; ##P<0.01 vs Model group;&P<0.05 vs 3-MA group.

图4 TTM可减轻PSCI大鼠脑组织中炎症反应,减少神经元凋亡

Fig.4 Trillium tschonoskii Maxim (TTM) attenuates inflammatory response and reduces neuronal apoptosis in the brain tissue of PSCI rats. A: Immunofluorescence staining of the neurons in the hippocampal tissue of the rats in each group (×40). B: Changes in the number of neuronal dendritic spines in each group (n=3, ×40). **P<0.01 vs Sham group; #P<0.05 vs Model group; &&P<0.01 vs 3-MA group.

图5 Bcl-2, Bax, IL-10, IL-1β和 TNF-α蛋白表达情况

Fig.5 Western blotting for detecting Bcl-2, Bax, IL-10, IL-1β, and TNF-α protein expressions in the hippocampal tissue of the rats in each group (n=3). *P<0.05, **P<0.01 vs Sham group; #P<0.05, ##P<0.01 vs Model group; &P<0.05, &&P<0.01 vs 3-MA group.

图6 TTM激活卒中后认知障碍大鼠脑组织内质网自噬

Fig.6 TTM activates ER-phagy in the brain tissue of rats with PSCI. A: Immunohistochemical staining for ATG5 and GRP78 in the hippocampal tissue of the rats in each group (×40). B: Immunofluorescence detection of FAM134B (green) , calnexin (green) and LC3 (red) expression (×40).

图7 FAM134B, GRP78, P62, ATG5和 LC3蛋白表达情况

Fig.7 Western blotting of FAM134B, GRP78, P62, ATG5, and LC3 protein expression in the hippocampal tissue of the rats in each group (n=3). *P<0.05, **P<0.01 vs Sham group; ##P<0.01 vs Model group; &P<0.05 vs 3-MA group.

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头顶一颗珠通过调节内质网自噬保护卒中后认知障碍大鼠的神经功能

Trillium tschonoskii Maxim saponin protects neurological function in rats with post-stroke cognitive impairment by promoting ER-phagy

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