[1]薛大力a,张晓哲,王 慧,等.通络益智方通过PERK介导的内质网应激途径减轻大鼠血管性痴呆模型记忆缺陷的实验研究[J].现代检验医学杂志,2025,40(01):17-23.[doi:10.3969/j.issn.1671-7414.2025.01.04]
　XUE Dalia,ZHANG Xiaozhe,WANG Hui,et al.Experimental Study of Tongluo Yizhi Formula Attenuating Memory Deficits in Rat Model of Vascular Dementia through PERK-mediated Endoplasmic Reticulum Stress Pathway[J].Journal of Modern Laboratory Medicine,2025,40(01):17-23.[doi:10.3969/j.issn.1671-7414.2025.01.04]
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通络益智方通过PERK介导的内质网应激途径减轻大鼠血管性痴呆模型记忆缺陷的实验研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年01期
页码:
17-23
栏目:
论著
出版日期:
2025-01-15

文章信息/Info

Title:
Experimental Study of Tongluo Yizhi Formula Attenuating Memory Deficits in Rat Model of Vascular Dementia through PERK-mediated Endoplasmic Reticulum Stress Pathway
文章编号:
1671-7414（2025）01-0017-07
作者:
薛大力1a; 张晓哲2; 王 慧2; 马冠峰1a; 郭素鹏1b
（1. 邢台医学高等专科学校第二附属医院 a．神经内科；b．检验科，河北邢台 054000；2. 邢台医学高等专科学校，河北邢台 054000）
Author(s):
XUE Dali1a; ZHANG Xiaozhe2; WANG Hui2; MA Guanfeng1a; GUO Supeng1b
(1a. Department of Neurology； 1b. Department of Clinical Laboratory，the Second Affiliated Hospital of Xingtai Medical College, Hebei Xingtai 054000，China；2. Xingtai Medical College，Hebei Xingtai 054000，China)
关键词:
血管性痴呆; 通络益智方; 内质网应激
分类号:
R-332；
DOI:
10.3969/j.issn.1671-7414.2025.01.04
文献标志码:
A
摘要:
目的 观察自拟通络益智方对血管性痴呆（VD）大鼠记忆缺陷及内质网应激（ERS）的影响及其机制。方法将50 只SPF 级Wistar 大鼠随机分为假手术（Sham）组、模型组、通络益智方（TLYZF）低剂量（12.5 mg/kg/d）组、TLYZF 中剂量（25 mg/kg/d）组和TLYZF 高剂量（50 mg/kg/d）组，每组10 只。除Sham 组外，其他组大鼠采用改良双侧颈总动脉结扎法构建VD 模型。TLYZF 干预4 周后，采用Morris 水迷宫实验评估大鼠学习记忆能力；苏木素- 伊红染色检测海马病理结构；尼氏染色检测海马神经元损伤；原位末端转移酶标记染色检测海马神经元凋亡；酶联免疫吸附试验检测海马组织炎性因子含量；蛋白免疫印迹法检测海马组织细胞凋亡及ERS 相关蛋白的表达。结果 与Sham 组比较，模型组大鼠平均逃逸潜伏期延长，穿越平台次数减少（t=14.059，8.534），海马神经元排列紊乱、形态模糊、胞质空泡化、胞核固缩、炎性细胞浸润增多，尼氏体数量减少（t=17.131），神经元凋亡率升高（t=17.701），TNF-α，IL-1β 和IL-6 含量增多（t=6.541，6.957，10.014），Bcl-2 相关X 蛋白（Bax）、裂解型半胱氨酸蛋白酶3（cleavedcaspase-3）、葡萄糖调控蛋白78（GRP78）、磷酸化内质网激酶（p-PERK/PERK）、磷酸化真核细胞起始因子2α（p-eIF2α）/ eIF2α，激活转录因子4（ATF4）和转录因子CCAAT/ 增强子结合蛋白同源蛋白（CHOP）表达增多（t=13.548 ～ 76.468），B 淋巴细胞瘤-2（Bcl-2）蛋白表达降低（t=39.691），差异具有统计学意义（均P<0.05）。与模型组比较，TLYZF 治疗后大鼠平均逃逸潜伏期缩短（t=2.476，7.266，11.306），穿越平台次数增多（t=2.187，4.471，6.932），海马神经元相对规则、细胞核清晰、炎性细胞浸润明显减少，尼氏体含量增加（t=4.359，9.477，11.449），神经元凋亡率降低（t=3.631，6.145，7.580），TNF-α，IL-1β 和IL-6 含量减少（t=2.382 ～ 8.293），Bax（t=4.696，16.250，20.250），cleavedcaspase-3（t=15.205，27.000，26.833），GRP78（t=5.918，13.139，13.741），p-PERK/PERK（t=13.416，14.230，17.889），p-eIF2α/ eIF2α（t=20.152，39.346，50.750），ATF4（t=12.093，24.395，22.946）和CHOP 蛋白（t=21.592，19.207，22.136）表达降低，Bcl-2 蛋白表达增多（t=7.474，20.761，35.350），差异具有统计学意义（均P<0.05），且高剂量组治疗效果最佳。结论 自拟通络益智方可减轻VD 大鼠海马神经元凋亡和炎症损伤，其作用机制可能与抑制PERK 介导的ERS 有关。
Abstract:
Objective To observe the effect of self-formulated Tongluo Yizhi formula on memory deficits and endoplasmic reticulum stress (ERS) in rats with vascular dementia (VD) and its mechanism. Methods Fifty SPF-grade Wistar rats were randomly divided into sham surgery group, model group, TLYZF low-dose (12.5 mg/kg/d) group, TLYZF medium dose (25 mg/ kg/d) group and TLYZF high-dose (50 mg/kg/d) group, with 10 rats in each group. Except for the Sham group, other rats were subjected to modified bilateral carotid artery ligation to construct VD models. After 4 weeks of TLYZF intervention, Morris water maze experiment was used to evaluate the learning and memory abilities of rats．Hematoxylin eosin staining was used to detect the pathological structure of the hippocampus．Nissl staining was used to detect hippocampal neuronal damage．In situ end transfer enzyme labeling staining was used to detect hippocampal neuronal apoptosis．Enzyme-linked immunosorbent assay was used to detect the content of inflammatory factors in hippocampal tissue．Protein immunoblotting was used to detect apoptosis of hippocampal tissue cells and the expression of ERS related proteins．Results Compared with the Sham group, the average escape latency of the model group was prolonged（t=14.059）, the number of crossing the platform was reduced（t=8.534）, the hippocampal neurons were disorderly arranged, the morphology was fuzzy, the cytoplasmic vacuolization, the nuclear pyknosis, the inflammatory cell infiltration increased, the number of Nissl bodies decreased（t=17.131）, and the neuronal apoptosis rate increased（t=17.701）. The contents of tumor necrosis factor-α (TNF-α), interleukin-1 β (IL-1β) and IL-6 increased（t=6.541，6.957，10.014）,the expression of Bcl-2 associated X protein (Bax), cleaved caspase-3, glucose-regulated protein 78 (GRP78), phosphorylated endoplasmic reticulum kinase (p-PERK) /PERK, phosphorylated eukaryotic initiation factor 2α (p-eIF2α)/eIF2α, activating transcription factor 4 (ATF4) and CCAAT/ enhancer binding protein (C/EBP) homologous protein (CHOP) increased（t=13.548 ～ 76.468）, while the expression of B-cell lymphoma-2 (Bcl-2) proteinwas decreased （t=39.691）, and the difference were statistically significant (all P < 0.05), respectively. Compared with the model group, the average escape latency of TLYZF-treated rats was shortened（t=2.476，7.266，11.306）, the number of platform crossing was increased（t=2.187，4.471，6.932）, the hippocampal neurons were relatively regular, the nucleus was clear, the inflammatory cell infiltration was significantly reduced, the content of Nissl bodies was increased（t=4.359，9.477，11.449），the apoptosis rate of neurons was decreased（t=3.631，6.145，7.580），the contents of TNF-α, IL-1β and IL-6 were decreased （t=2.382 ～ 8.293），the expression of Bax（t=4.696，16.250，20.250）, cleaved caspase-3（t=15.205，27.000， 26.833）, GRP78（t=5.918，13.139，13.741）, p-PERK/PERK（t=13.416，14.230，17.889）, p-eIF2α/eIF2α（t=20.152， 39.346，50.750）, ATF4（t=12.093，24.395，22.946）and CHOP protein(（t=21.592，19.207，22.136）decreased，while Bcl-2 protein increased（t=7.474，20.761，35.350）,the differences were statistically significant (all P<0.05),respectively , and the high-dose group had the best therapeutic effect.Conclusion Self-formulated Tongluo Yizhi formula could reduce the apoptosis and inflammatory injury of hippocampal neurons in VD rats, and its mechanism may be related to inhibiting ERS mediated by PERK.

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备注/Memo

备注/Memo:
基金项目：河北省医学科学研究课题计划（20242376）。
作者简介：薛大力（1981-），男，硕士，副教授/ 副主任医师，研究方向：神经内科诊疗，E-mail：xuedaliluck5733@163.com。
通讯作者：张晓哲（1980-），女，硕士，副教授，研究方向：中医药研究，E-mial：zhangxiaozhe123@126.com。
更新日期/Last Update: 2025-01-15