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郑钰,张逸凡,王子焱,刘承鑫,任欣,聂子星,郭志华,申思.血塞泰对缺血性脑卒中大鼠STIM1、Orai1蛋白表达的影响[J].湖南中医药大学学报,2024,44(4):557-564[点击复制] |
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| 血塞泰对缺血性脑卒中大鼠STIM1、Orai1蛋白表达的影响 |
| 郑钰,张逸凡,王子焱,刘承鑫,任欣,聂子星,郭志华,申思 |
| (湖南中医药大学第一中医临床学院, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一中医临床学院, 湖南 长沙 410007;湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学第一中医临床学院, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 研究血塞泰对缺血性脑卒中（ischemic stroke，IS）大鼠基质相互作用分子1（stromal interaction molecule 1，STIM1）及钙释放激活钙通道蛋白1（calcium release-activated calcium channel protein 1，Orai1）的影响。方法 选取60只SPF级雄性SD大鼠，随机分为假手术组10只、造模组50只，采用线栓法制备大脑中动脉栓塞（medial cerebral artery occlusion，MCAO）模型。采用随机数字表法将造模成功的41只大鼠分为模型组、血塞泰低剂量组（12.6 g/kg）、血塞泰中剂量组（25.2 g/kg）、血塞泰高剂量组（50.4 g/kg）和阿司匹林肠溶片组（18 mg/kg），其中血塞泰低剂量组9只，其余每组各8只；假手术组和模型组给予等量生理盐水灌胃。每组连续干预7 d。采用改良神经功能损伤评分（modified Neurological Severity Score，mNSS）评估各组大鼠神经功能损伤情况，TTC染色法检测大鼠脑组织的梗死面积，ELISA法测定大鼠血清中的白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）含量，RT-PCR检测脑组织中STIM1、Orai1 mRNA的表达，免疫共沉淀检测脑组织STIM1、Orai1的蛋白相对表达水平。结果 与假手术组相比，模型组大鼠mNSS评分增加（P＜0.01），脑梗死率增加（P＜0.01），血清IL-6、TNF-α含量增加（P＜0.01），脑组织STIM1、Orai1 mRNA表达水平和蛋白相对表达水平升高（P＜0.01）。与模型组相比，血塞泰各剂量组和阿司匹林肠溶片组大鼠mNSS评分下降（P＜0.05），血清IL-6、TNF-α含量下降（P＜0.01），脑组织STIM1、Orai1 mRNA表达水平下降（P＜0.01）；血塞泰中、高剂量组和阿司匹林肠溶片组大鼠脑梗死率下降（P＜0.01），脑组织STIM1、Orai1的蛋白相对表达水平降低（P＜0.05，P＜0.01）。与阿司匹林肠溶片组相比，血塞泰低剂量组脑梗死率增加（P＜0.01），血清IL-6、TNF-α含量增加（P＜0.01），脑组织STIM1、Orai1 mRNA表达水平增加（P＜0.05）；血塞泰中剂量组血清IL-6、TNF-α含量明显增加（P＜0.01），脑组织STIM1、Orai1 mRNA表达水平下降（P＜0.05，P＜0.01）；血塞泰高剂量组脑梗死率下降（P＜0.01），血清IL-6、TNF-α含量明显下降（P＜0.01），脑组织STIM1、Orai1 mRNA表达水平明显下降（P＜0.01）。与血塞泰低、中剂量组相比，血塞泰高剂量组mNSS评分下降（P＜0.05，P＜0.01），脑梗死率下降（P＜0.01），血清IL-6、TNF-α含量下降（P＜0.01），脑组织STIM1、Orai1 mRNA表达水平下降（P＜0.01）。结论 血塞泰能够抑制STIM1、Orai1的表达及结合，减轻炎症反应，改善MCAO大鼠神经功能缺损与降低神经元损伤，从而发挥抗IS的作用。 |
| 关键词: 缺血性脑卒中 血塞泰 基质相互作用分子1 钙释放激活钙通道蛋白1 炎症 |
| DOI：10.3969/j.issn.1674－070X.2024.04.005 |
| 投稿时间：2023-11-27 |
| 基金项目:国家自然科学基金（81673955）；湖南省卫生健康委科研计划项目（202203074016）；湖南省研究生科研创新项目（CX20230832）。 |
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| ffects of Xuesaitai on protein expressions of STIM1 and Orai1 in ischemic stroke rats |
| ZHENG Yu,ZHANG Yifan,WANG Ziyan,LIU Chengxin,REN Xin,NIE Zixing,GUO Zhihua,SHEN Si |
| (The First Clinical School of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Clinical School of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;The First Clinical School of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To study the effects of Xuesaitai on expressions of stromal interaction molecule 1(STIM1) and calcium release-activated calcium channel protein 1(Orai1) in ischemic stroke (IS) rats.Methods A total of 60 SPF-grade male SD rats were randomized into sham-operated group (n=10) and model group (n=50).A middle cerebral artery occlusion (MCAO) model was prepared by thread embolism.A total of 41 successfully modeled rats were randomly subdivided into model group,low-,medium-,and high-dose (12.6 g/kg,25.2 g/kg,50.4 g/kg) Xuesaitai groups,and enteric-coated aspirin tablets group (18 mg/kg),with nine rats in low-dose Xuesaitai group and eight rats in each of the other groups.The sham-operated and model groups were given an equal volume of normal saline by gavage for continuous seven days.The modified Neurological Severity Score (mNSS) was used to assess the neurological damage in rats from each group,TTC staining was used to determine the infarct area of rat brain tissue,ELISA was used to measure the levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in rat serum,RT-PCR was used to check the mRNA expressions of STIM1 and Orai1 in brain tissue,and immunoprecipitation method was used to determine the relative protein expression levels of STIM1 and Orai1 in the brain tissue.Results Compared with the sham-operated group,the model group showed increased mNSS scores (P＜0.01),enlarged infarct area (P＜0.01),elevated serum levels of IL-6 and TNF-α(P＜0.01),and higher mRNA and relative protein expression levels of STIM1 and Orai1 in the brain tissue (P＜0.01).Compared with the model group,the rats in each dose group of Xuesaitai as well as enteric-coated aspirin tablets group showed decreased mNSS scores (P＜0.05),reduced serum levels of IL-6 and TNF-α(P＜0.01),and decreased mRNA expression levels of STIM1 and Orai1 in the brain tissue (P＜0.01).Additionally,the rats in the medium-and high-dose Xuesaitai groups,and enteric-coated aspirin tablets group exhibited decreased infarct area (P＜0.01) and reduced relative protein expression levels of STIM1 and Orai1 in the brain tissue (P＜0.05,P＜0.01).The high-dose Xuesaitai group exhibited lower cerebral infarct rate (P＜0.01),significantly reduced serum levels of IL-6 and TNF-α(P＜0.01),and significantly decreased mRNA expression levels of STIM1 and Orai1 in the brain tissue (P＜0.01).Compared with the low-and medium-dose Xuesaitai groups,the high-dose Xuesaitai group exhibited reduced mNSS scores (P＜0.05,P＜0.01),lower cerebral infarct rate (P＜0.01),reduced serum levels of IL-6 and TNF-α(P＜0.01),and decreased mRNA expression levels of STIM1 and Orai1 in the brain tissue (P＜0.01).Conclusion Xuesaitai can inhibit the expression and binding of STIM1 and Orai1,alleviate the inflammatory response,relieve neurological deficits,and reduce neuronal damage in MCAO rats,thus exerting an anti-IS effect. |
| Key words: ischemic stroke Xuesaitai stromal interaction molecule 1 calcium release-activated calcium channel protein 1 inflammation |
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