论文部分内容阅读
该研究通过观察冰片是否具有促进梓醇、葛根素透过局灶性脑缺血模型大鼠血脑屏障的作用,及其是否与激活β_2肾上腺素受体-eNOS-NO途径有关,从医学生物学作用角度阐释冰片“通窍引经”的传统功效及其机制。该研究采用大鼠大脑中动脉阻塞局灶性脑缺血模型,5 d后神经功能障碍评分达5分者为造模成功。将模型成功大鼠随机分为7组,用药前经统计学检验各组间无显著性差异后,灌服给予相应药物[梓醇45 mg·kg~(-1)、葛根素200 mg·kg~(-1)、冰片200 mg·kg~(-1)、布他沙明(Butoxamine,BTX)1.5 mg·kg~(-1)]:模型组(M组,溶剂)、梓葛组(ZG组)、梓葛冰组(ZGB组)、梓冰组(ZB组)、葛冰组(GB组)、β_2受体阻断剂+梓葛组(BTX+ZG组)、β_2受体阻断剂+梓葛冰组(BTX+ZGB组),另设假手术组(S组,溶剂),每组10只。各组给药10 min后取材,采用UPLC-MS测定脑脊液中梓醇和葛根素含量,Western blot法测定脑组织中eNOS含量、β_2受体表达,Griess法检测脑组织中NO含量。结果显示,与假手术组相比,造模大鼠神经功能得分均值均在5分以上,具有显著差异(P<0.05),显示造模成功。除假手术组外,经单因素方差分析,给药前各组之间的神经功能评分均无显著性差异。给药后与模型组相比,ZG组梓醇质量分数为26.673μg·L~(-1),葛根素含量在检测限以下,脑组织中eNOS表达和NO含量、β_2受体表达没有显著差异;与ZG组相比,ZGB组、ZB组、GB组脑脊液中梓醇质量分数增加至40~43μg·L~(-1)、葛根素质量分数增加至72~75μg·L~(-1),脑组织中eNOS表达和NO含量、β_2受体表达均有显著升高(P<0.05),显示冰片有促进作用;与ZG组相比,BTX+ZG组大鼠脑脊液中梓醇质量分数为21.401μg·L~(-1),葛根素检测不出,2组无显著差异,显示BTX无抑制作用;与ZGB组相比,BTX+ZGB组脑脊液中梓醇质量分数32.826μg·L~(-1)、葛根素质量分数47.820μg·L~(-1),脑组织中eNOS表达和NO含量均有显著下降(P<0.05),显示BTX有抑制作用。该研究说明冰片能够显著促进梓醇、葛根素透过局灶性脑缺血大鼠血脑屏障,其作用可被β_2肾上腺素受体阻断剂布他沙明所阻断,其机制可能与上调β_2肾上腺素受体从而促进eNOS表达,进而使NO含量升高有关。
The study by observing whether or not borneol promotes the action of catalpol and puerarin on the blood-brain barrier in focal cerebral ischemia model rats and whether it is related to the activation of β 2 -adrenoceptor-eNOS-NO pathway, The Role of Learning to Explain the Traditional Function and Mechanism of Borneol “Tong Qiao Yin Jing”. This study used a model of middle cerebral artery occlusion in rats with focal cerebral ischemia. After 5 days, the score of neurological dysfunction reached 5 points was successful. The successful model rats were randomly divided into 7 groups. Before statistical analysis, there was no significant difference among the groups. After administration of the corresponding drugs [catalpol 45 mg · kg -1, puerarin 200 mg · kg -1 (-1), borneol 200 mg · kg -1, butoxamine 1.5 mg · kg -1]: model group (M group, solvent), Zi Ge group ZG group, ZGB group, Zixing group, GB group, β_2 receptor blocker group, BTX + ZG group, β_2 receptor blocker Disinfectant + Zi Gebing group (BTX + ZGB group), the other a sham operation group (S group, solvent), each group of 10. The contents of catalpol and puerarin in cerebrospinal fluid were measured by UPLC-MS. The eNOS content and β 2 receptor expression in brain tissue were measured by Western blot, and the content of NO in brain tissue was detected by Griess method. The results showed that compared with the sham-operated group, the mean scores of neurological function of the model rats were all above 5 points (P <0.05), indicating successful modeling. In addition to the sham-operated group, there was no significant difference in neurological function scores among the groups before administration after single-factor analysis of variance. Compared with model group, the content of catalpol in ZG group was 26.673μg · L -1 after administration, the content of puerarin was below the detection limit, the expression of eNOS and NO, and the expression of β 2 receptor in brain tissue were not significantly different ; Compared with ZG group, the contents of catalpol in cerebrospinal fluid of ZGB group, ZB group and GB group increased to 40 ~ 43μg · L -1 and the content of puerarin increased to 72 ~ 75μg · L -1, , ENOS expression, NO content and β 2 receptor expression in brain tissue were significantly increased (P <0.05), indicating that borneol has a promoting effect. Compared with ZG group, the content of catalpol in CSF of BTX + ZG group was 21.401μg · L -1, puerarin was not detected, no significant difference between the two groups, showing no inhibitory effect of BTX; Compared with ZGB group, BTX + ZGB group cerebrospinal fluid catalpol concentration 32.826μg · L ~ ( -1), puerarin content 47.820μg · L -1, eNOS expression and NO content in brain tissue were significantly decreased (P <0.05), indicating that BTX has inhibitory effect. This study indicates that borneol can significantly promote catalpol and puerarin through the blood-brain barrier of focal cerebral ischemia rats, which can be blocked by β 2 adrenergic receptor antagonist bursalpine, which may be related to Upregulation of β_2 adrenergic receptor to promote eNOS expression, thereby increasing the NO content.