木犀草素增强亚剂量卡托普利降压作用并减少咳嗽的研究

发布时间：2019-05-15 22:11

【摘要】：背景和目的:血管紧张素转化酶抑制剂(ACEI)在抗高血压和保护靶器官方面的效应已得到公认,但其缺点是引起刺激性干咳的发生率高,因此,降低ACEI类药物咳嗽发生率具有重要的临床意义。鉴于木犀草素不但具有降压作用,而且还具有较强的抗炎作用,能降低气道高反应性等药理活性,本研究拟评估木犀草素能否增强亚剂量卡托普利降压作用并减少其咳嗽的不良反应,并探讨相互作用机制。实验方法:1)构建肾性高血压大鼠模型,检测大鼠尾动脉血压变化。2)二氧化硫引咳法研究对小鼠咳嗽的影响。3)雾化吸入卵白蛋白建立气道高反应性小鼠模型,ELISA法测定支气管肺泡灌洗液(BALF)中IL-4、IL-5及IFN-γ的含量。4)MTT法检测小鼠巨噬细胞代谢活力。5)ELISA法和NO检测试剂盒检测药物对巨噬细胞分泌IL-1 β、IL-10、TNF-a和NO的影响。6)激光共聚焦扫描显微镜检测药物对LPS诱导小鼠巨噬细胞内Ca2+浓度和pH变化的影响。实验结果:1)高血压大鼠分组给药并检测血压,结果显示,从第7天开始,给予卡托普利和木犀草素的动物血压明显下降。第14天,亚剂量卡托普利和木犀草素联用时降压效果基本达到常剂量卡托普利的效果。2)小鼠咳嗽随卡托普利用量增加而增加,联用木犀草素可以降低小鼠咳嗽发生率。3)木犀草素明显降低高气道反应性小鼠模型BALF中IL-4和IL-5水平,卡托普利无明显抑制作用。两者亚剂量联用能抑制IL-4和IL-5分泌,升高IFN-γ水平。4)木犀草素明显抑制巨噬细胞代谢活力,降低TNF-a,IL-1β和IL-10分泌水平,而卡托普利在高浓度时有明显抑制作用,低浓度卡托普利则无明显抑制作用。两者联用能抑制TNF-a,IL-1β和IL-10的分泌,无协同效应,但有一定相加效应。另外卡托普利和木犀草素均能抑制NO的分泌,但也无协同效应。5)木犀草素和卡托普利明显降低胞内Ca2+浓度,升高胞内pH,但两者联用对胞内Ca2+浓度和pH变化无明显协同效应。结论:1)亚剂量木犀草素和亚剂量卡托普利联用基本能达到常剂量卡托普利的降压效果。2)亚剂量卡托普利和木犀草素联用能减少前者导致的小鼠咳嗽发生率,其机制可能与卡托普利剂量下降以及木犀草素能抑制IL-4、IL-5分泌,增加IFN-丫水平,调节TH1和TH2细胞比例有关。3)体外卡托普利和木犀草素联用虽无明显协同抗炎作用,但有一定相加效应,木犀草素的抗炎作用对减少卡托普利咳嗽发生频率可能有帮助,其抗炎机制可能通过降低细胞内Ca2+浓度、升高pH值、抑制巨噬细胞分泌促炎细胞因子等功能有关。
[Abstract]:Background & objective: the effect of angiotensin converting enzyme inhibitor (ACEI) on antihypertension and protection of target organs has been recognized, but its disadvantage is that the incidence of irritating dry cough is high, so, It is of great clinical significance to reduce the incidence of cough with ACEI drugs. In view of the fact that luteolin not only has antihypertensive effect, but also has strong anti-inflammatory effect, which can reduce airway hyperresponsiveness and other pharmacological activities. The purpose of this study was to evaluate whether luteolin can enhance the antihypertensive effect of subdose captopril and reduce the adverse reactions of cough, and to explore the mechanism of interaction. Methods: 1) the rat model of renal hypertension was established, and the changes of blood pressure in tail artery of rats were detected. 2) the effect of sulfur dioxide on cough in mice was studied. 3) the model of airway hyperresponsiveness was established by atomization inhalation of ovalbumin. The contents of IL-4,IL-5 and IFN- 纬 in (BALF) of bronchoalveolar lavage fluid were determined by ELISA. 4) the metabolic activity of mouse macrophages was detected by MTT. 5) ELISA assay and NO kit were used to detect the secretion of IL-1 尾 by macrophages. The effects of IL-10,TNF-a and NO were detected by confocal laser scanning microscope. 6) the effects of drugs on the concentration of Ca2 and the change of pH in mouse macrophages induced by LPS were detected by laser confocal scanning microscope. The results were as follows: 1) Hypertension rats were divided into groups and blood pressure was measured. The results showed that the blood pressure of animals given captopril and rhinocerol decreased significantly from the 7th day. On the 14th day, the antihypertensive effect of subdose captopril combined with luteolin basically reached the effect of constant dose captopril. 2) the cough of mice increased with the increase of captopril dosage. Combined with lignogenin could reduce the incidence of cough in mice. 3) luteolin significantly decreased the levels of IL-4 and IL-5 in BALF model of hyperairway reactive mice, but captopril had no significant inhibitory effect. The combination of the two subdoses could inhibit the secretion of IL-4 and IL-5 and increase the level of IFN- 纬. 4) the metabolic activity of macrophages and the secretion of TNF-a,IL-1 尾 and IL-10 were significantly decreased by luteolin. Captopril had obvious inhibitory effect at high concentration, but no significant inhibitory effect at low concentration of captopril. The combination of them could inhibit the secretion of TNF-a,IL-1 尾 and IL-10 without synergistic effect, but had a certain additive effect. In addition, captopril and luteolin could inhibit the secretion of NO, but there was no synergistic effect. 5) luteolin and captopril significantly decreased intracellular Ca2 concentration and increased intracellular pH,. However, there was no significant synergistic effect on intracellular Ca2 concentration and pH. Conclusion: 1) the combination of subdose luteolin and subdose captopril can basically achieve the antihypertensive effect of constant dose captopril. 2) the combination of subdose captopril and luteolin can reduce the incidence of cough in mice caused by the former. The mechanism may be related to the decrease of captopril dose and the inhibition of IL-4,IL-5 secretion and the increase of IFN- level. Regulating the proportion of TH1 and TH2 cells is related. 3) although the combination of captopril and luteolin in vitro has no obvious synergistic anti-inflammatory effect, it has a certain additive effect. The anti-inflammatory effect of luteolin may be helpful to reduce the frequency of captopril cough. The anti-inflammatory mechanism may be related to the decrease of intracellular Ca2 concentration, the increase of pH value and the inhibition of macrophage secretion of pro-inflammatory cytokines.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R544.1

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