川芎嗪对野百合碱诱导大鼠肺动脉高压的影响及机制

目的 观察川芎嗪(TMP)对野百合碱(MCT)诱导的肺动脉高压(PAH)大鼠模型的作用,并探讨相关机制。 方法 36只雄性SD大鼠随机分为对照组、模型组、TMP治疗组(TMP组),每组12只。模型组和TMP组在第1天给予腹腔注射MCT 50 mg/kg,建立PAH模型,28 d后TMP组给予川芎嗪80 mg/kg灌胃,每天1次,连续4周。实验结束后测大鼠右心室收缩压(RVSP)、右心室肥厚指数(RVHI);取右肺中叶行苏木素-伊红(HE)和Masson染色,计算管壁厚度百分比(WT%)和管壁面积百分比(WA%);透射电镜观察肺组织结构;免疫组化法和Western blotting检测肺组织中Hippo信号通路相关蛋白YAP1、WWTR1、TEAD1表达水平。 结果 与对照组相比,模型组大鼠RVSP、RVHI升高(P<0.001,P<0.001),肺中小动脉管腔狭窄,胶原纤维增生,肺血管YAP1、WWTR1、TEAD1蛋白表达水平升高(P=0.004,P<0.001,P=0.013)。与模型组相比,TMP组大鼠RVSP、RVHI降低(P=0.001,P<0.001),肺动脉重构及右室肥厚情况有所改善,纤维化程度减轻,肺组织YAP1、WWTR1、TEAD1蛋白表达水平降低(P=0.045,P=0.011,P=0.034)。相较于对照组,TMP组RVSP、RVHI升高(P=0.045,P=0.045),YAP1、WWTR1、TEAD1蛋白表达水平上升(P=0.080,P=0.013,P=0.468),但仅有WWTR1的蛋白表达水平差异有统计学意义。提示川芎嗪作用有限。 结论 TMP可对MCT诱导的PAH大鼠模型产生作用,其机制可能与抑制Hippo信号通路蛋白表达有关。

Effects and mechanism of tetramethylpyrazine on monocrotaline induced pulmonary arterial hypertension in rats

Objective To investigate the effects and mechanism of tetramethylpyrazine(TMP)on monocrotaline(MCT)induced pulmonary arterial hypertension(PAH)in rats. Methods A total of 36 male SD rats were randomly divided into control, model and TMP groups, with 12 rats in each group. The rats in the model and TMP groups were given intraperitoneal injection of MCT 50 mg/kg on the 1st day to establish PAH model. After 28 days, the TMP group was given tetramethylpyrazine 80 mg/kg by gavage once a day for 4 weeks. At the end of the experiment, the right ventricular systolic pressure(RVSP)was detected and right ventricular hypertrophy index(RVHI)was counted; the pathomorphological changes of lung tissue were observed with hematoxylin-eosin(HE)staining and Masson staining, and the percentage of wall thickness(WT%)and wall area(WA%)were calculated. The structure of lung tissue was observed with a transmission electron microscope. The expressions of Hippo signal pathway related proteins YAP1, WWTR1 and TEAD1 in lung tissue were detected with immunohistochemical staining and Western blotting. Results Compared with the control group, the model group had increased RVSP and RVHI(P<0.001, P<0.001), significantly narrowed lumen of pulmonary arteries, proliferated collagen fibers, and increased protein expressions of YAP1, WWTR1 and TEAD1(P=0.004, P<0.001, P=0.013). Compared with the model group, the TMP group had decreased RVSP and RVHI(P=0.001, P<0.001), improved pulmonary artery remodeling and right ventricular hypertrophy, reduced fibrosis, and decreased protein expressions of YAP1, WWTR1 and TEAD1(P=0.045, P=0.011, P=0.034). Compared with the control group, the TMP group had increased RVSP and RVHI(P=0.045, P=0.045), and increased protein expressions of YAP1, WWTR1 and TEAD1(P=0.080, P=0.013, P=0.468), in which, the difference of WWTR1 protein expression was significant, suggesting that TMP has limited effects. Conclusion TMP has effects on MCT-induced PAH rat model. The underlying mechanism may be related to the inhibition of expressions of Hippo signal pathway related proteins.