Suppr超能文献

腺病毒介导的靶向细胞质磷脂酶 A2α 的 RNA 干扰减轻小鼠局灶性脑缺血损伤。

Adenoviruses-mediated RNA interference targeting cytosolic phospholipase A2α attenuates focal ischemic brain damage in mice.

机构信息

Department of Neurology, Cangzhou Central Hospital, Cangzhou, Hebei 061001, P.R. China.

出版信息

Mol Med Rep. 2018 Apr;17(4):5601-5610. doi: 10.3892/mmr.2018.8610. Epub 2018 Feb 15.

Abstract

Cerebral ischemia injury is a clinical, frequently occurring disease, which causes a heavy burden on society and families. It has been demonstrated that cytosolic phospholipase A2α (cPLA2α) is significant in neurological injury caused by ischemic brain injury, and inhibition of cPLA2α may reduce stroke injury. In the present study, the role of cPLA2α was investigated in a mouse model of middle cerebral artery occlusion and/or reperfusion (MCAO/R) using an effective cPLA2α inhibitor and adenoviruses‑mediated RNA interference. The most effective recombinant adenovirus encoding cPLA2α small interfering RNA (pAd‑siRNA‑cPLA2α) was constructed and selected. MCAO/R surgery is used to construct the model of focal ischemic brain damage in mice. Adenoviruses‑mediated RNA interference targeting cPLA2α was administered by stereotactic surgery 2 h before the MCAO/R. The expression/activity of cPLA2α and cPLA2α‑derived injurious lipid mediators was assessed. pAd‑siRNA‑cPLA2α‑treated animals (RNA interference; RNAi group) were compared with pAd-siRNA-control-treated animals (negative group) with regard to neurological deficit, motor function, pathological changes, apoptosis, and infarct volume. The RNAi group animals reduced the expression level of cPLA2α, as determined by western blotting and reverse transcription‑quantitative polymerase chain reaction, the improvement of locomotor function was evaluated by rotarod test, and the decrease of apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end‑labeling staining. The decreased infarct areas were evaluated by 2,3,5‑triphenyltetrazolium chloride staining. The expression levels of prostaglandin E2, leukotrienes B4, lysophosphatidylcholine and free fatty acids were reduced in the RNAi group when compared with the negative control group. Thus, the data indicates that the expression level of cPLA2α was effectively controlled by pAd‑siRNA‑cPLA2α treatment. pAd‑siRNA‑cPLA2α treatment, in reducing the levels of inflammatory factors, neurological deficit and tissue damage, represents an effective potential therapeutic strategy. pAd‑siRNA‑cPLA2α reduces cPLA2α expression levels with long‑term efficacy, thereby improving functional deficits and effectively attenuating ischemic brain damage. Thus, pAd‑siRNA‑cPLA2α shows potential value for therapeutic evaluation in ischemic brain damage.

摘要

脑缺血损伤是一种临床常见疾病,给社会和家庭带来沉重负担。已有研究表明,胞质型磷脂酶 A2α(cPLA2α)在缺血性脑损伤引起的神经损伤中起重要作用,抑制 cPLA2α 可能减轻中风损伤。本研究采用有效的 cPLA2α 抑制剂和腺病毒介导的 RNA 干扰,在大脑中动脉闭塞和/或再灌注(middle cerebral artery occlusion and/or reperfusion,MCAO/R)的小鼠模型中探讨 cPLA2α 的作用。构建并筛选出最有效的编码 cPLA2α 小干扰 RNA 的重组腺病毒(pAd-siRNA-cPLA2α)。采用立体定向手术在 MCAO/R 前 2 h 给予腺病毒介导的靶向 cPLA2α 的 RNA 干扰。评估 cPLA2α 及其衍生的损伤性脂质介质的表达/活性。用 pAd-siRNA-cPLA2α 处理的动物(RNA 干扰;RNAi 组)与用 pAd-siRNA 对照处理的动物(阴性组)进行比较,比较指标包括神经功能缺损、运动功能、病理变化、细胞凋亡和梗死体积。Western blot 和逆转录定量聚合酶链反应检测到 RNAi 组动物 cPLA2α 的表达水平降低,旋转棒试验评估运动功能改善,末端脱氧核苷酸转移酶 dUTP 缺口末端标记染色评估细胞凋亡减少。2,3,5-三苯基四氮唑氯化物染色评估梗死面积减少。与阴性对照组相比,RNAi 组前列腺素 E2、白三烯 B4、溶血磷脂胆碱和游离脂肪酸的表达水平降低。因此,数据表明 pAd-siRNA-cPLA2α 治疗可有效控制 cPLA2α 的表达水平。pAd-siRNA-cPLA2α 治疗可降低炎症因子水平、神经功能缺损和组织损伤,是一种有效的潜在治疗策略。pAd-siRNA-cPLA2α 通过降低 cPLA2α 的表达水平来改善功能缺陷,并有效减轻缺血性脑损伤。因此,pAd-siRNA-cPLA2α 在缺血性脑损伤的治疗评价中具有潜在价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9e10/5866000/f45686d6d3b1/MMR-17-04-5601-g00.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验