基于内质网应激信号探讨熄风解痉汤对蛛网膜下腔出血大鼠早期脑损伤的保护作用  

Protective Effect of Xifeng Jiejing Tang on Early Brain Injury in Rats with Subarachnoid Hemorrhage Based on Endoplasmic Reticulum Stress Signaling Pathway

作　　者：向兴刚[1] 周益凡 牛淑亮[2] 依马木·依达依吐拉[1] 赵永[1] 买买江·阿不力孜[1] 林琳[1] XIANG Xinggang;ZHOU Yifan;NIU Shuliang;Yimamu·Yidayitula;ZHAO Yong;Maimaijiang·Abulizi;LIN Lin(Affiliated Hospital of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830000,Xinjiang,China;School of Basic Medical Science,Xinjiang Medical University)

机构地区：[1]新疆医科大学附属中医医院,乌鲁木齐830000 [2]新疆医科大学基础医学院

出　　处：《中西医结合心脑血管病杂志》2025年第8期1165-1171,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：新疆维吾尔自治区自然科学基金项目(No.2021D01C230)。

摘　　要：目的:基于内质网应激信号通路探讨熄风解痉汤(XFJJT)对蛛网膜下腔出血(SAH)大鼠早期脑损伤(EBI)的保护作用机制。方法:将48只无特定病原体(SPF)级SD大鼠随机分为假手术组(Sham组)、模型组(SAH组)、XFJJT低剂量组和XFJJT高剂量组,每组12只。视交叉前池注射自体血法构建SAH大鼠模型,XFJJT低剂量组、XFJJT高剂量组大鼠每12 h接受1次灌胃给药,剂量分别为每次5、10 mL/kg,连续给药3 d。Sham组和SAH组大鼠作为对照,给予相同体积的0.9%氯化钠溶液灌胃。采用Longa's 5级标准评分法评估大鼠的神经功能缺损程度,干/湿重法进行脑组织含水量测定,脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)实验检测神经元细胞凋亡率。苏木精-伊红染色法观察脑组织的病理变化。Evans蓝外渗的量检测血脑屏障通透性。酶联免疫吸附实验(ELISA)法检测大鼠外周血清中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的表达水平。蛋白免疫印迹法(Western Blot)检测内质网应激相关的葡萄糖调节蛋白78(GRP78)、转录因子C/EBP同源蛋白(CHOP)、含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-12的表达水平。结果:与Sham组比较,SAH组大鼠神经功能缺损评分、脑组织含水量、脑神经元凋亡率及IL-1β、IL-6和TNF-α表达水平升高,脑组织中GRP78、CHOP、Caspase-12表达水平升高,差异均有统计学意义(P<0.05)。与SAH组比较,给药3 d后XFJJT低剂量组及XFJJT高剂量组大鼠神经功能缺损评分、脑组织含水量、脑神经元凋亡率及IL-1β、IL-6和TNF-α表达水平下降,GRP78、CHOP、Caspase-12表达水平下降,差异均有统计学意义(P<0.05)。结论:XFJJT通过调控GRP78、CHOP、Caspase-12蛋白的表达水平,减轻SAH后的内质网应激反应,发挥保护SAH大鼠大脑的作用,减轻SAH后EBI。Objective:To investigate the protective effect of Xifeng Jiejing Tang(XFJJT)on early brain injury(EBI)after subarachnoid hemorrhage(SAH)in rats.Methods:Forty-eight SPF-grade SD rats were randomly divided into sham-operated group,SAH group,XFJJT low-dose group,and XFJJT high-dose group,with 12 rats in each group.SAH rat model was constructed by injecting autologous blood into the anterior pool of the optic chiasm.The rats in the XFJJT low-dose and XFJJT high-dose groups received the drug by gavage once every 12 h,with doses of 5 mL/kg and 10 mL/kg,respectively.The drug was administered for 3 consecutive days.The rats in the sham-operated group and the SAH group were given the same volume of 0.9%NaCl solution by gavage as a control.The rats were evaluated for neurological deficit symptoms using Longa's 5-grade standardized scoring method,the water content of brain tissue was determined by the dry/wet weight method,the apoptosis rate of neuronal cells was detected by TUNEL assay,the pathological changes of brain tissue were observed by hematoxylin-eosin staining,and the permeability of the blood-brain barrier was detected by the amount of Evans blue extravasation.Additionally,the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the peripheral blood of the rats were detected by enzyme-linked immunosorbent assay(ELISA).And the expression levels of endoplasmic reticulum stress-associated glucose-regulated protein 78(GRP78),transcription factor C/EBP homologous protein(CHOP),and cysteine-containing aspartic acid protease(Caspase)-12 were detected by Western blot.Results:Compared with the sham-operation group,rats in the SAH group showed elevated neurological deficit scores,brain tissue water content,apoptosis rate of brain neurons,and expression levels of IL-1β,IL-6,and TNF-α,as well as increased expression of GRP78,CHOP,and Caspase-12 in brain tissue,with significant and statistically significant differences(P<0.05).Compared with the SAH group,At 3 days after administration

关 键 词：蛛网膜下腔出血 早期脑损伤 熄风解痉汤 内质网应激 实验研究 

分 类 号：R285.5[医药卫生—中药学]