目的:筛选氧化应激状态下CHCHD2(coiled-coil-helix-coiled-coil-helix domain-containing 2)的互作蛋白,以期挖掘出CHCHD2保护神经细胞对抗氧化应激损伤的潜在机制。方法:通过Lipofectamine 2000分别在人神经母细胞瘤细胞系SH-SY5Y中转染含有Flag标签的对照质粒及CHCHD2过表达质粒,用100μmol/L过氧化叔丁醇(tert-butyl hydroperoxide,TBHP)或蒸馏水处理24 h后,采用免疫共沉淀(CoIP)的方法富集各组细胞中与CHCHD2相结合的蛋白,SDS-PAGE跑浓缩胶,切取条带,胶内酶解后进行液相色谱-质谱联用(LC-MS/MS)分析、数据库检索及生物信息分析,筛选与CHCHD2互作的蛋白,并对功能进行初步分析。结果:(1)CHCHD2具有保护SH-SY5Y细胞对抗TBHP诱导的氧化应激损伤作用;(2)CoIP-MS结果提示,不同于生理状态,在氧化应激状态下共有64个蛋白是与CHCHD2互作的特有差异表达蛋白(differentially expressed proteins,DEPs);(3)通过GO功能注释和KEGG富集分析,我们发现氧化应激状态下特有DEPs主要在外泌体和细胞浆中发挥作用,参与蛋白翻译及翻译起始等生物过程,在蛋白及poly(A)RNA结合方面发挥分子功能,并参与糖代谢过程;(4)DEPs还参与了负性调控活性氧生物合成过程和对过氧化氢的反应等抗氧化应激相关生物过程,其中肿瘤坏死因子受体相关蛋白1(tumor necrosis factor receptor-associated protein 1,TRAP1)和热休克蛋白家族D成员1(heat shock protein family D member1,HSPD1)是抗氧化应激过程中重要的候选蛋白;(5)通过蛋白互作网络分析,我们发现在氧化应激状态下特异性存在3个蛋白[Y盒结合蛋白1(Y-box-binding protein 1,YBX1)、含TCP1分子伴侣亚基6A(chaperonin containing TCP1 subunit 6A,CCT6A)和细胞色素C氧化酶装配因子4同源物(cytochrome C oxidase assembly factor 4 homolog,COA4)]与CHCHD2有直接相互作用,也是后续需要重点关注的蛋白。结论:利用CoIP-MS法成功筛选出生理状态及氧化应激状态下CHCHD2的互作蛋白,并挖掘出与其抗氧化应激过程密切相关的2个候选蛋白(TRAP1和HSPD1)及另外3个与其直接作用的候选蛋白(YBX1、CCT6A和COA4),为进一步深入探索CHCHD2抗氧化应激作用中的生物过程及分子机制奠定基础。
在缺血性卒中的多种风险原因中,高脂血症特别是高胆固醇血症始终占有着很关键的地位,通过降低血浆中胆固醇特别是低密度脂蛋白胆固醇的水平,可以很有效地减少缺血性卒中发生的风险,这一观点已是世界神经科专家的普遍共识。他汀类药物是缺血性卒中二级预防的基石,被广泛应用于临床,然而,他汀类药物也有其自身的局限性。首先,如果他汀类药物剂量增加一倍,其降脂效果仅增加6%,这使得许多患者仅靠他汀类药物很难达到LDL-C目标。其次,有些患者不能耐受他汀类药物,这一现象在接受大剂量他汀类药物治疗的中国患者中尤为明显。近几年新上市的新型降脂药物–前蛋白转化酶枯草溶菌素9 (proprotein convertase subtilisin/kexin type 9, PCSK9)抑制剂则为缺血性卒中的治疗提供了新的策略。Among the myriad risk factors for ischemic stroke, hyperlipidemia, particularly hypercholesterolemia, consistently holds a pivotal role. The reduction of plasma cholesterol levels, notably low-density lipoprotein cholesterol (LDL-C), has been recognized as an effective strategy to mitigate the risk of ischemic stroke. This perspective has garnered widespread acceptance among neurology experts globally. Statins, as the mainstay of secondary prevention for ischemic stroke, are extensively utilized in clinical settings. However, statins are not without their inherent limitations. Firstly, the lipid-lowering efficacy of statins plateaus with increased dosage;doubling the dose results in a mere 6% increase in cholesterol reduction, making it challenging for many patients to achieve their LDL-C targets with statin monotherapy. Secondly, statin intolerance is not uncommon, with a pronounced incidence observed among Chinese patients undergoing high-dose statin therapy. In this context, the advent of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors, a novel class of lipid-lowering agents, offers a promising alternative for the management of ischemic stroke, presenting a new therapeutic horizon in the field.
卒中后抑郁是一种常见的卒中并发症,对患者的康复和生活质量有着显著影响。约三分一卒中患者伴有糖尿病,而糖尿病可加剧卒中后抑郁的风险,两者之间在病理生理学上存在相互促进的关系。本文将探讨糖尿病与卒中后抑郁之间的病理生理学联系,并从药物治疗的角度进行分析,旨在为合并糖尿病的卒中后抑郁患者提供预防和诊治参考。Post-stroke depression is a common complication of stroke, significantly affecting patients’ recovery and quality of life. Approximately one-third of stroke patients are diagnosed with diabetes, and diabetes can exacerbate the risk of post-stroke depression. There is a mutually reinforcing relationship between the two in terms of pathophysiology. This article will explore the pathophysiological connection between diabetes and post-stroke depression and analyze it from the perspective of drug therapy, aiming to provide prevention and treatment references for patients with post-stroke depression who also have diabetes.
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