背景:早产是一个高发病率和高死亡率的重大的全球性健康问题.白质损伤是早产儿最常见的脑损伤.丹参是一种传统的草药,在亚洲国家最常用于治疗心脑血管疾病. 目的:探讨丹参在早产儿脑白质损伤方面的治疗作用. 方法:选取3 d龄新生雄性SD大鼠18 只,随机分为正常组、白质损伤组、白质损伤+丹参组,后两组通过永久性结扎右侧颈总动脉建立早产白质损伤大鼠模型,正常组不造模.白质损伤+丹参组自造模第1天起给予丹参5 mg/(kg·d)腹腔注射,正常组及白质损伤组给予同剂量PBS进行干预,连续给药7 d.造模14 d后处死大鼠,采用苏木精-伊红染色显微镜下观察大鼠脑组织病理情况,免疫荧光法观察髓鞘碱性蛋白和CC1在脑组织中的表达水平,应用液相色谱-质谱联用仪分析丹参发挥作用的可能途径. 结果与结论:①白质损伤组大鼠胼胝体结构不规则,细胞肿胀坏死,髓鞘形成显著减少,神经纤维排列不规则且松散,髓鞘显著减少;与白质损伤组相比,白质损伤+丹参组大鼠脑组织细胞肿胀减轻,损伤减轻,髓鞘增多;②髓鞘碱性蛋白与髓鞘的形成密切相关,CC1是髓鞘少突胶质细胞的标志物,白质损伤组大鼠脑组织中髓鞘碱性蛋白及CC1表达显著降低;白质损伤+丹参组大鼠脑组织中髓鞘碱性蛋白及CC1的表达显著增加(P<0.000 1),表明丹参减轻了白质损伤;③液相色谱-质谱联用仪分析发现白质损伤+丹参组和白质损伤组之间差异最显著的蛋白与补体和凝血级联反应有关;④研究表明丹参具有治疗早产白质损伤的潜在运用价值.
BACKGROUND:Premature birth is a major global health problem associated with high mortality and morbidity.White matter injury is the most common brain injury in preterm infants.Salvia miltiorrhiza is a traditional herbal plant that is commonly used to treat cardiovascular and cerebrovascular diseases in Asian countries. OBJECTIVE:To investigate the therapeutic effect of Salvia miltiorrhiza on white matter injury in preterm infants. METHODS:Eighteen neonatal male Sprague-Dawley rats at 3-day gestational age were selected and randomized into normal group,white matter injury group,and Salvia miltiorrhiza group.Animal models of preterm white matter injury were established by permanent ligation of the right common carotid artery in the latter two groups.Rats in the Salvia miltiorrhiza group were given intraperitoneal injection of Salvia miltiorrhiza(5 mg/kg·d)for 7 consecutive days.Normal group and white matter injury group were given the same volume of PBS for intervention.On the 14th day after modeling,the rats were sacrificed.Brains were pathologically observed by hematoxylin-eosin staining under microscope,and the expression levels of myelin basic protein and CC1 in brain tissue were visualized using immunofluorescence.Furthermore,liquid chromatography-tandem mass spectrometry was used to analyze possible pathways for the action of Salvia miltiorrhiza. RESULTS AND CONCLUSION:In the white matter injury group,the structure of the corpus callosum was irregular and the cells appeared swollen and necrotic.In addition,induction of white matter injury resulted in significantly reduced myelin formation,with irregular and loosely arranged nerve fibers and significantly decreased myelin sheaths.Interestingly,white matter injury rats treated with Salvia miltiorrhiza had reduced cellular swelling,reduced lesions,and increased myelin sheaths.The expression of myelin basic protein was closely related to myelin formation,and CC1 was a marker of myelin oligodendrocytes.Salvia miltiorrhiza significantly up-regulated the expressions of myelin basic protein and CC1 in white matter injury rats(P<0.000 1),indicating that Salvia miltiorrhiza alleviated white matter injury.Liquid chromatography-tandem mass spectrometry analysis showed that the therapeutic effect of Salvia miltiorrhiza in the rat model of white matter injury was closely related to the regulation of complement and coagulation cascades.To conclude,Salvia miltiorrhiza may be a potential therapeutic agent for treating preterm white matter injury.