Introduction: Culturing cerebrovascular smooth muscle cells (CVSMCs) in vitro can provide a model for studying many cerebrovascular diseases. This study describes a convenient and efficient method to obtain mouse CVSMCs by enzyme digestion. Methods: Mouse circle of Willis was isolated, digested, and cultured with platelet-derived growth factor-BB (PDGF-BB) to promote CVSMC growth, and CVSMCs were identified by morphology, immunofluorescence analysis, and flow cytometry. The effect of PDGF-BB on vascular smooth muscle cell (VSMC) proliferation was evaluated by cell counting kit (CCK)-8 assay, morphological observations, Western blotting, and flow cytometry. Results: CVSMCs cultured in a PDGF-BB-free culture medium had a typical peak-to-valley growth pattern after approximately 14 days. Immunofluorescence staining and flow cytometry detected strong positive expression of the cell type-specific markers alpha-smooth muscle actin (α-SMA), smooth muscle myosin heavy chain 11 (SMMHC), smooth muscle protein 22 (SM22), calponin, and desmin. In the CCK-8 assay and Western blotting, cells incubated with PDGF-BB had significantly enhanced proliferation compared to those without PDGF-BB. Conclusion: We obtained highly purified VSMCs from the mouse circle of Willis using simple methods, providing experimental materials for studying the pathogenesis and treatment of neurovascular diseases in vitro. Moreover, the experimental efficiency improved with PDGF-BB, shortening the cell cultivation period.
We evaluated the effect of acute hypobaric hypoxia (AHH) on the hippocampal region of the brain in early-stage spontaneously hypertensive male rats. The rats were classified into a control (ground level; ~ 400 m altitude) group and an AHH experimental group placed in an animal hypobaric chamber at a simulated altitude of 5500 m for 24 h. RNA-Seq analysis of the brains and hippocampi showed that differentially expressed genes (DEGs) were primarily associated with ossification, fibrillar collagen trimer, and platelet-derived growth factor binding. The DEGs were classified into functional categories including general function prediction, translation, ribosomal structure and biogenesis, replication, recombination, and repair. Pathway enrichment analysis revealed that the DEGs were primarily associated with relaxin signaling, PI3K-Akt signaling, and amoebiasis pathways. Protein–protein interaction network analysis indicated that 48 DEGs were involved in both inflammation and energy metabolism. Further, we performed validation experiments to show that nine DEGs were closely associated with inflammation and energy metabolism, of which two ( Vegfa and Angpt2 ) and seven ( Acta2, Nfkbia, Col1a1, Edn1, Itga1, Ngfr , and Sgk1 ) genes showed up and downregulated expression, respectively. Collectively, these results indicated that inflammation and energy metabolism-associated gene expression in the hippocampus was altered in early-stage hypertension upon AHH exposure.
哌托生特作为一种组胺H3受体拮抗剂/反向激动剂,其明确的促觉醒作用以及低药物依赖性使其迅速成为欧美治疗成人发作性睡病的一线用药.本文通过总结组胺H3受体分布及作用特点,阐明了哌托生特的作用靶点及其产生的相关效应;并通过总结哌托生特的药物化学特点以及将其与常见抗疲劳药物成瘾性之间进行对比,展望了哌托生特在特殊环境/条件下的应用潜能.本文为哌托生特在我国特殊环境/条件下,如军事、轮班、紧急任务等方面的合理化应用提供了理论依据.
目的 探究服用10 mg扎来普隆对主观视觉功能的影响,评估服药后的安全放飞时间.方法 15名健康男性被试者,采用双盲、单模拟和自身交叉对照研究方法,交叉服用10 mg扎来普隆和安慰剂(前后相隔3 d),服药后被试者均给予了 2.5 h的睡眠时间,于服药3、4、5 h后分别进行一次主观视觉功能评估.评估内容包括视力、主观快速暗适应、夜间视力、对比敏感度、立体视觉以及临界闪光融合频率.结果 服药3 h后扎来普隆组和安慰剂组0.6 c/d空间频率下对比敏感度为(15.49±2.92)dB vs(16.51±2.03)dB(P=0.035),差异虽有统计学意义,但其结果仍在正常范围内,扎来普隆并未导致该服药时段对比敏感度异常.服药5h后主观暗适应时间为(3.63±0.85)s vs(4.37±1.14)s(P=0.038),服用扎来普隆缩短了暗适应时间;服药后3、4、5 h扎来普隆组和安慰剂组的左、右眼视力,主观暗适应、夜间视力,对比敏感度、立体视觉以及临界闪光融合频率指标均无统计学差异(P>0.05).结论 服用10 mg扎来普隆自服药3 h起对主观视觉功能无不良影响.