脑栓通预防寒潮促发高血压大鼠卒中的机制：差异蛋白质组学研究

Mechanisms of Naoshuantong capsule in the prevention of cold wave-induced stroke in hypertensive rats： a differential proteomics study

目的探讨脑栓通胶囊对人工寒潮促发高血压大鼠卒中的预防作用和机制。方法130只雄性SD大鼠随机分为假手术组（n＝30）、模型对照组（n＝50）和脑栓通治疗组（n＝50；脑栓通灌胃0.5 g/kg，2/d共7 d）。应用双肾双夹法制作大鼠肾血管性高血压模型，在造模12周后进行人工寒潮处理3 d（温度22 ℃、光照12 h，温度4 ℃、黑暗12 h，共3个循环）。在实验终点分别提取脑组织标本，采用差异蛋白组学技术对差异表达的蛋白质点进行鉴定、功能分类和初步分析，采用蛋白质印迹分析对部分关键蛋白质进行验证。结果假手术组大鼠未发生卒中，模型对照组卒中发生率（36.00%，18/50）显著高于脑栓通治疗组（18.00%，9/50；χ2＝4.110，P＝0.043）。双向凝胶电泳图谱从14个差异蛋白质点鉴定得到了6种差异表达蛋白质，其中表达上调的超氧化物歧化酶2（superoxide dismutase 2, SOD2）和表达下调的B细胞淋巴瘤因子10（B-cell lymphoma 10, Bcl-10）处于多种蛋白质相互作用的中心位置，并通过蛋白质印迹分析得到验证。 结论脑栓通治疗能显著降低人工寒潮诱发的肾血管性高血压大鼠卒中发生率。SOD2上调和Bcl-10下调可能参与了其预防寒潮促发高血压性大鼠卒中的机制。

ObjectiveTo investigate the preventive effect and mechanisms of Naoshuantong capsule in stroke induced by artificial cold wave in hypertensive rats.MethodsA total of 130 SD rats were randomly divided into a sham operation group （n=30）, a model control group （n=50） and a Naoshuantong treatment group （n=50; intragastric administration of Naoshuantong, 0.5 g/kg, 2/d for 7 days）. Renovascular hypertensive rats model were established by two-kidney, two clip method. At 13th week after operation, rats were exposed to artificial cold wave for 3 days （12 h light of 22 ℃ and 12 h dark of 4 ℃, 3 cycles）. The brain tissue samples were extracted at the end of the experiment. Differential protein proteomic techniques were used for the identification, functional classification and preliminary analysis of the differentially expressed protein spots, and Western blot was used for the validation of some key proteins.ResultsThere was no occurrence of stroke in the sham operation group, and the incidence of stroke in the model control group （36.00%, 18/50） was significantly higher than that in the Naoshuantong treatment group （18.00%, 9/50; χ2=4.110, P=0.043）. With the two-dimensional gel electrophoresis analysis, 6 different proteins were identified from 14 protein spots. Among them, the up-regulated superoxide dismutase 2 （SOD2） and the down-regulated B-cell lymphoma 10 （Bcl-10） were found to be at the central location of protein interactions, which has been verified by Western blot.ConclusionNaoshuantong can reduce the occurrence of stroke induced by artificial cold wave in renovascular hypertensive rats. SOD2 up-regulating and Bcl-10 down-regulating may be involved in the mechanisms of of Naoshuantong in the prevention of cold wave-induced stroke in hypertensive rats.