Synthesis of Novel Aryloxyethylamine Derivatives and Evaluation of Their in Vitro and in Vivo Neuroprotective Activities

A series of aryloxyethylamine derivatives were designed, synthesized and evaluated for their biological activity. Their structures were confirmed by ¹H‐NMR, ¹³C‐NMR, FT‐IR and HR‐ESI‐MS. The preliminary screening of neuroprotection of compounds in vitro was detected by MTT, and the anti‐ischemic activity in vivo was tested using bilateral common carotid artery occlusion in mice. Most of these compounds showed potential neuroprotective effects against the glutamate‐induced cell death in differentiated rat pheochromocytoma cells (PC12 cells), especially for (4‐fluorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}(4‐methoxyphenyl)methanone, (4‐bromophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐chlorophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone, (4‐chlorophenyl)(1‐{2‐[(naphthalen‐2‐yl)oxy]ethyl}piperidin‐4‐yl)methanone, (4‐chlorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone and {1‐[2‐(4‐bromophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone, which exhibited potent protection of PC12 cells at three doses (0.1, 1.0, 10 μM). Compounds (4‐fluorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, (4‐fluorophenyl){1‐[2‐(naphthalen‐2‐yloxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}(4‐methoxyphenyl)methanone and {1‐[2‐(4‐chlorophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone possessed the significant prolongation of the survival time of mice subjected to acute cerebral ischemia and decreased the mortality rate at all five doses tested (200, 100, 50, 25, 12.5 mg/kg) and had significant neuroprotective activity. In addition, (4‐fluorophenyl){1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}methanone, {1‐[2‐(4‐methoxyphenoxy)ethyl]piperidin‐4‐yl}(4‐methoxyphenyl)methanone and {1‐[2‐(4‐chlorophenoxy)ethyl]piperidin‐4‐yl}(4‐chlorophenyl)methanone possessed outstanding neuroprotection in vitro and in vivo. These compounds can be used as a promising neuroprotective agents for future development of new anti‐ischemic stroke agents. Basic structure–activity relationships are also presented.     

Multifrequency Microwave Imaging for Brain Stroke Detection

CT and MRI are often used in the diagnosis and monitoring of stroke. However, they are expensive, time-consuming, produce ionizing radiation (CT), and not suitable for continuous monitoring stroke. Microwave imaging (MI) has been extensively investigated for identifying several types of human organs, including breast, brain, lung, liver, and gastric. The authors recently developed a holographic microwave imaging (HMI) algorithm for biological object detection. However, this method has difficulty in providing accurate information on embedded small inclusions. This paper describes the feasibility of the use of a multifrequency HMI algorithm for brain stroke detection. A numerical system, including HMI data collection model and a realistic head model, was developed to demonstrate the proposed method for imaging of brain tissues. Various experiments were carried out to evaluate the performance of the proposed method. Results of experiments carried out using multifrequency HMI have been compared with the results obtained from single frequency HMI. Results showed that multifrequency HMI could detect strokes and provide more accurate results of size and location than the single frequency HMI algorithm.     

Melatonin against acute ischaemic stroke dependently via suppressing both inflammatory and oxidative stress downstream signallings

This study tested the hypothesis that melatonin (Mel) therapy preserved the brain architectural and functional integrity against ischaemic stroke (IS) dependently through suppressing the inflammatory/oxidative stress downstream signalling pathways. Adult male B6 (n = 6 per each B6 group) and TLR4 knockout (ie TLR4^?/?) (n = 6 per each TLR4^?/? group) mice were categorized into sham control (SC^B6), SC^TLR4?/?, IS^B6, IS^TLR4?/?, IS^B6 + Mel (i.p. daily administration) and IS^TLR4?/? + Mel (i.p. daily administration). By day 28 after IS, the protein expressions of inflammatory (HMBG1/TLR2/TLR4/MAL/MyD88/RAM TRIF/TRAF6/IKK‐α/p‐NF‐κB/nuclear‐NF‐κB/nuclear‐IRF‐3&7/IL‐1β/IL‐6/TNF‐α/IFN‐γ) and oxidative stress (NOX‐1/NOX‐2/ASK1/p‐MKK4&7/p‐JNK/p‐c‐JUN) downstream pathways as well as mitochondrial‐damaged markers (cytosolic cytochrome C/cyclophilin D/SRP1/autophagy) were highest in group IS^B6, lowest in groups SC^B6 and SC^TLR4?/?, lower in group IS^TLR4?/? + Mel than in groups IS^TLR4?/? and IS^B6 + Mel and lower in group IS^B6 + Mel than in group IS^TLR4?/? (all P < .0001). The brain infarct volume, brain infarct area and the number of inflammatory cells in brain (CD14/F4‐88) and in circulation (MPO+//Ly6C+/CD11b+//Ly6G+/CD11b+) exhibited an identical pattern, whereas the neurological function displayed an opposite pattern of inflammatory protein expression among the six groups (all P < .0001). In conclusion, TLR inflammatory and oxidative stress signallings played crucial roles for brain damage and impaired neurological function after IS that were significantly reversed by Mel therapy.     

阿替普酶联合依达拉奉治疗急性缺血性卒中的疗效及神经功能缺损与时间窗的关系研究

目的:探讨阿替普酶联合依达拉奉治疗急性缺血性卒中的疗效及神经功能缺损与时间窗的关系。方法:选取大连医科大学附属大连市中心医院于2016年3月~2018年10月间收治的急性缺血性卒中患者117例,根据随机数字表法将患者分为对照组(n=58,阿替普酶治疗)和研究组(n=59,阿替普酶联合依达拉奉治疗),比较两组患者临床疗效、神经功能缺损情况、基质金属蛋白酶-9(MMP-9)、白介素-6(IL-6)水平、头颅CT梗死面积,观察两组治疗期间不良反应发生情况。结果:研究组的总有效率为84.75%(50/59),高于对照组的63.79%(37/58)(P<0.05)。两组患者治疗2周后MMP-9、IL-6、美国国立卫生研究院卒中量表(NIHSS)评分均较治疗前降低,且研究组低于对照组(P<0.05)。研究组治疗24h、48h、72h的头颅CT梗死面积小于对照组(P<0.05)。治疗后研究组发病72h内、发病48h内患者NIHSS评分、头颅CT梗死面积高于发病24h内,且发病72h内高于发病48h内(P<0.05)。两组患者不良反应发生率比较无差异(P>0.05)。结论:阿替普酶联合依达拉奉治疗急性缺血性卒中,疗效确切,可有效改善患者过氧化损伤,降低细胞因子水平,且越早的时间窗内接受治疗的患者,其神经功能缺损、脑梗死面积改善效果越好。     

不同病情急性缺血性脑卒中患者血清过氧化还原蛋白1、钙调蛋白、触珠蛋白水平与预后的关系分析

摘要 目的：探讨不同病情急性缺血性脑卒中（AIS）患者血清过氧化还原蛋白1(PRDX1)、钙调蛋白(CAM)、触珠蛋白(HPT)水平与预后的关系。方法：收集2018年4月~2019年4月期间本院收治的127例AIS患者为研究对象,根据患者的病情分为轻度组(39例)、中度组(48例)、重度组(40例),根据患者的改良Rankin 量表( mRS)评分将患者分为预后良好组(73例)和预后不良组(54例),另选同期在我院进行健康检查的健康受试者50例为对照组。对比所有受试者PRDX1、CAM、HPT水平。对比不同预后患者的一般资料、PRDX1、CAM、HPT水平。分析PRDX1、CAM、HPT与NIHSS评分、mRS评分的关系。以多因素Logistic回归分析AIS患者预后的影响因素。结果：轻度组、中度组、中度组的PRDX1、CAM、HPT水平均高于对照组,且随着患者的病情加重,PRDX1、CAM、HPT水平依次升高,差异均有统计学意义(P<0.05)。预后不良组患者的年龄、PRDX1、CAM、HPT均高于预后良好组(P<0.05)。经Pearson检验,AIS患者的PRDX1、CAM、HPT与NIHSS、mRS评分均呈正相关(P<0.05)。经多因素Logistic回归分析可得,年龄较高、PRDX1水平升高、CAM水平升高、HPT水平升高是AIS患者预后不良的危险因素(P<0.05)。结论：AIS患者的PRDX1、CAM、HPT水平异常升高,且与其病情及预后呈现明显的相关性,年龄、PRDX1、CAM、HPT是患者预后的影响因素,对于病情的评估、预后的判断有一定的临床指导价值。     

吡拉西坦联合鼠神经生长因子对急性缺血性脑卒中患者的疗效及Hcy、PCT和皮质醇水平的影响

目的:探讨吡拉西坦联合鼠神经生长因子对急性缺血性脑卒中患者的疗效及对同型半胱氨酸(Hcy)、降钙素原(PCT)和皮质醇水平的影响。方法:回顾性分析我院2017年2月～2018年11月收治的73例急性缺血性脑卒中患者为研究对象,依据入院先后顺序分为对照组(n=35)和观察组(n=38)。对照组患者采用吡拉西坦治疗,观察组基于对照组加以鼠神经生长因子治疗。观察并比较两组临床疗效,治疗前后美国国立卫生研究院卒中量表(NIHSS)、日常生活能力(ADL),治疗前及治疗2周结束时用全自动生化分析仪测定血浆Hcy水平,用放射免疫学分析法测定PCT水平,用化学发光法测定皮质醇水平,用超声多普勒诊断仪测定基底动脉、左右大脑中动脉血流。结果:治疗后,观察组NIHSS评分低于对照组(8.96±1.21)vs(11.27±1.59)分,ADL评分高于对照组(74.21±9.75)vs(66. 04±8.03)分(P0.05)。观察组总有效率高于对照组89.47%vs 68.57%(P0.05)。治疗后,观察组Hcy、PCT及皮质醇水平低于对照组(14.27±2.01)vs (18.51±2.84)μmol/L、(0.25±0.03)vs (0.31±0.04)μg/L、(171.93±23.86)vs(228.75±30.27)nmol/L(P0.05)。治疗后,观察组脑血流学指标高于对照组基底动脉(43.81±6.84)vs(39.62±5.27)mL/min、左大脑中动脉血流(64.27±9.95)vs(57.03±7.52)mL/min、右大脑中动脉血流(62.85±9.01)vs(56.64±7.42)mL/min(P0.05)。结论:吡拉西坦联合鼠神经生长因子能够提高急性缺血性脑卒中的疗效,降低Hcy、PCT及皮质醇水平,促进神经功能的恢复。     

加味星蒌承气汤对急性缺血性脑卒中患者神经功能及血脂、血液流变学的影响

目的:探讨加味星蒌承气汤对急性缺血性脑卒中患者神经功能及血脂、血液流变学的影响。方法:选取2017年8月～2019年6月期间我院收治的急性缺血性脑卒中患者96例,将入选患者根据随机数字表法分为对照组(n=48)和研究组(n=48),对照组患者予以常规西医治疗,研究组患者在对照组基础上联合加味星蒌承气汤治疗,对比两组患者疗效、神经功能及血脂、血液流变学情况,记录两组患者治疗期间不良反应情况。结果:研究组治疗8 d后的临床总有效率为91.67%(44/48),显著高于对照组患者的72.92%(35/48)(P0.05)。两组治疗8 d后加拿大神经功能评分量表(CNS)、美国国立卫生研究所卒中量表(NIHSS)评分均下降(P0.05),且研究组低于对照组(P0.05)。两组治疗8 d后总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、全血黏度高切、全血黏度低切、血浆黏度、纤维蛋白原均下降(P0.05),且研究组低于对照组(P0.05);高密度脂蛋白胆固醇(HDL-C)升高(P0.05),且研究组高于对照组(P0.05)。两组不良反应发生率比较无明显差异(P0.05)。结论:急性缺血性脑卒中患者采用加味星蒌承气汤治疗,疗效显著,可有效改善患者神经功能及血脂、血液流变学,且安全性较好。     

Retracted: Upregulated microRNA-31 inhibits oxidative stress-induced neuronal injury through the JAK/STAT3 pathway by binding to PKD1 in mice with ischemic stroke

Ischemic stroke (IS), which is characterized by high morbidity, disability, and mortality, is recognized as a major cerebrovascular disease. MicroRNA-31 (miR-31) was reported to participate in the progression of brain disease. The present study was conducted in order to investigate the effect of miR-31 on oxidative stress-induced neuronal injury in IS mice with the involvement of protein kinase D1 (PKD1) and the JAK/STAT3 pathway. C57BL/6J mice were used to establish the middle cerebral artery occlusion (MCAO) model. Astrocytes were transfected with miR-31 mimic, miR-31 inhibitor, si-PKD1, or JAK-STAT3 pathway inhibitor. Following the establishment of an oxygen–glucose deprivation (OGD) model, the astrocytes were cocultured with neuronal OGD. Lower miR-31, higher PKD1 expressions, and activated JAK/STAT3 pathway were found in both the MCAO and OGD models. miR-31 could negatively target PKD1. In an MCAO model, overexpressing miR-31 and silencing PKD1 reduced neuronal injury, cerebral infarct volume, neuron loss, and oxidative stress injury, inhibited the activation of JAK/STAT3 pathway and the expressions of PKD1, interleukin (IL)-1β, IL-6, tumor necrosis factor-α, malondialdehyde, 4-HNE, 8-HOdG, caspase-3, and Bax, but increased the superoxide dismutase content. In the OGD model, overexpression of miR-31 and silencing of PKD1 attenuated oxidative stress-induced neuronal injury, and diminished the lactate dehydrogenase leakage and reactive oxygen species level, accompanied by elevated neuronal viability. These results indicate that miR-31 alleviates inflammatory response as well as an oxidative stress-induced neuronal injury in IS mice by downregulating PKD1 and JAK/STAT3 pathway.     

Retracted: Upregulation of acetylcholinesterase caused by downregulation of microRNA-132 is responsible for the development of dementia after ischemic stroke

MicroRNA-132 (miR-132) has been shown to participate in many diseases. This study aimed to understand the correlation between the level of miR-132 and the severity of dementia post-ischemic stroke. An online tool ( www.mirdb.org ) was used to find the miR-132 binding site in acetylcholinesterase (ACHE) 3′-untranslated region (UTR), followed by a luciferase reporter assay to validate ACHE as a miR-132 target. A similar relationship between miR-132 and ACHE was also established in cerebrospinal fluid samples collected from human subjects. A negative correlation was established between ACHE and miR-132 by measuring the relative luciferase activity. Meanwhile, Western blot analysis and real-time polymerase chain reaction were also conducted to compare the levels of ACHE messenger RNA and protein between two groups (dementia positive, n = 26 and dementia negative, n = 26) or among cells treated with miR-132 mimics, ACHE small interfering RNA, and miR-132 inhibitors. As shown in the results, miR-132 can reduce the expression of ACHE. Further experiments were also carried out to study the effect of miR-132 and ACHE on cell viability and apoptosis, and the results demonstrated that miR-132 enhanced cell viability while suppressing apoptosis. In addition, ACHE reduced cell viability while promoting apoptosis. miR-132 targeted ACHE and suppressed its expression. Additionally, miR-132 and ACHE have been shown to affect the cell viability and apoptosis in the central nervous system.     

缺血再灌卒中后抑郁小鼠模型的建立

目的探讨并建立卒中后抑郁小鼠动物模型。方法先利用结扎颈总动脉造成小鼠不完性全脑缺血再灌注病理损伤,利用悬尾和强迫游泳方法诱导小鼠抑郁模型,利用正交试验设计全面考察其影响因素,确定最佳造模条件。结果以缺血5min,再灌10min,再缺血5min所诱导抑郁模型最为明显,且死亡率较低。结论利用此方法所造卒中后抑郁模型具有操作简便、成功率较高且经济的特点,适用于该类药物筛选。