益气化痰方调节高血压大鼠脑血管适应性功能及炎症小体的研究

Study on Qi-Replenishing and Phlegm-Resolving Recipe Regulating Adaptive Function of Cerebral Vessels and Inflammasomes in Hypertension Rats

【目的】观察益气化痰方(QPR)对高血压大鼠脑血管适应性调节功能的影响。【方法】在实验前1周,QPR组和血管紧张素受体拮抗剂(ARB)组SD大鼠分别预先灌胃益气化痰方和替米沙坦,其余大鼠灌胃相同容量的磷酸盐缓冲溶液(PBS),设为PBS组。大鼠麻醉后,经颞窗持续灌流林格氏液,当平均动脉压(MAP)稳定时,测量基线大脑血流(CBF)值。分别经静脉泵入内皮素1(ET-1)或者血管紧张素Ⅱ(AngⅡ)制作2种高血压模型,以单独泵入林格氏液作为空白对照组。分别通过触须刺激或者局部灌流乙酰胆碱诱发功能性充血,再次测量CBF值。CBF的变化以相对于基线水平的升高百分比表示。采用蛋白免疫分析(Western Blot)法检测各组大鼠主动脉组织中的Nod样受体热蛋白结构域相关蛋白3(NLRP3)和剪切型白细胞介素1β(IL-1β)蛋白的表达。【结果】泵入AngⅡ后,大鼠CBF上升幅度较泵入林格氏液组明显降低(P<0.05)。AngⅡ+QPR组、AngⅡ+ARB组CBF上升幅度均高于单纯泵入AngⅡ组(P<0.05)。泵入ET-1后,大鼠CBF上升幅度较泵入林格氏液组降低(P<0.05),QPR组、ET-1+BQ123组CBF上升幅度均高于单纯泵入ET-1组(P<0.05)。以乙酰胆碱灌流检测脑血管内皮依赖性的充血反应,也获得类似的治疗结果。Western Blot结果显示,与PBS组比较,QPR组、ARB组大鼠主动脉组织的NLRP3和剪切型IL-1β蛋白的表达水平均明显下调(P<0.05)。【结论】QPR具有拮抗大脑皮层功能性充血反应和脑血管内皮依赖性充血反应的作用,可改善高血压大鼠的脑循环适应性调节功能,其作用机制可能与阻断炎症小体激活有关。

Objective To observe the effect of Qi-Replenishing and Phlegm-Resolving Recipe(QPR)on cerebral vessel-regulated adaptive function in rats with hypertension. Methods One week before surgery,the rats in QPR group and angiotensin Ⅱ(AngⅡ)receptor blocker(ARB)group were beforehand given intragastric administration of QPR and Telmisartan, respectively, and the other rats were given intragastric administration of phosphate buffer solution(PBS), served as PBS group. The value of cerebral blood flow(CBF) in anesthetized rats persistently perfused with Ringer’s solution through tempus window was monitored just at stabilization of mean arterial pressure(MAP). The 2 kinds of hypertension models were established by intravenous infusion pump with endothelin(ET-1)or AngⅡ,and the vehicle control group was set up by intravenous infusion of Ringer’s solution alone. And then CBF value was measured again after functional hyperemia induced by vibrissa stimulation or local perfusion with acetylcholine. CBF change was expressed as percentage of CBF increase relative to baseline level.The expression levels of Nod-like receptor protein 3(NLRP3)and cleaved interleukin 1 beta(IL-1β)in aorta tissue was determined by Western blotting assay. Results The increase amplitude of CBF in rats infused with AngⅡwas lower than that in the vehicle control group(P<0.05). The increase amplitude of CBF in the AngⅡ+ QPR group and AngⅡ+ ARB group was higher than that in the AngⅡalone infusion group(P<0.05). The increase amplitude of CBF in rats infused with ET-1 was lower than that in the vehicle control group(P<0.05). The increase amplitude of CBF in the ET-1 + QPR group and ET-1 + BQ123 group was higher than that in the ET-1 alone infusion group(P<0.05). However, the treatment results for functional hyperemia induced by local perfusion of acetylcholine were also obtained,being similar to the above results. Western Blotting results showed that the expression levels of NLRP3 and cleaved IL-1β in aorta tissue in QPR group and ARB group were reduced as compared with those in PBS group(P<0.05). Conclusion QPR has effects on resisting cerebrocortical functional hyperemia response and cerebrovascula endothelial-dependent hyperemia response,and ameliorating adaptive regulation of cerebral circulation in hypertension rats,the mechanism possible is related with blocking activation of inflammasome.