丹参酮ⅡA通过介导PI3K/Akt-eNOS信号通路改善野百合碱所致大鼠肺动脉高压  

Tanshinone ⅡA alleviates monocrotaline-induced pulmonary hypertension in rats through the PI3K/Akt-eNOS signaling pathway

作　　者：张喜民 刘思佳[1,2] 孙亚彬 李国锋[1,2,3] ZHANG Ximin;LIU Sijia;SUN Yabin;LI Guofeng(Department of Pharmacy,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Rational Medication Evaluation and Drug Delivery Technology Lab,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Guangdong Provincial Key Laboratory of New Drug Screening,School of Pharmaceutical Sciences,Southern Medical University,Guangzhou 510515,China;Guangzhou Civil Aviation College,Guangzhou 510403,China;Hutchison Whampoa Guangzhou Baiyunshan Chinese Medicine Co.,Ltd,Guangzhou 510515,China)

机构地区：[1]南方医科大学南方医院药学部,广东广州510515 [2]南方医科大学南方医院合理用药评价与药物递送发展实验室,广东广州510515 [3]南方医科大学药学院广东省新药筛选重点实验室,广东广州510515 [4]广州民航职业技术学院,广东广州510403 [5]广州白云山和记黄埔中药有限公司,广东广州510515

出　　处：《南方医科大学学报》2022年第5期718-723,共6页Journal of Southern Medical University

基　　金：广州市科技计划项目珠江科技新星专项(201806010066);广东省自然科学基金(2020A1515010396)。

摘　　要：目的 探究丹参酮ⅡA治疗野百合碱所致大鼠肺动脉高压(PAH)的作用机制。方法 选取100只SD雄性大鼠,按随机数字表法分为5组(20只/组):模型组(MCT)、丹参酮ⅡA组(TⅡA)、丹参酮ⅡA+PI3K抑制剂组(TP)、PI3K抑制剂组(PI)、对照组。除对照组颈背部注射等体积生理盐水外,剩余大鼠均颈背部注射野百合碱(MCT)诱导肺动脉高压模型。2周后,丹参酮ⅡA组腹腔注射丹参酮ⅡA10 mg/kg进行治疗,丹参酮ⅡA+PI3K抑制剂组注射丹参酮ⅡA10 mg/kg+PI3K抑制剂1 mg/kg,PI3K抑制剂组注射PI3K抑制剂1 mg/kg,对照组和模型组均腹腔注射等体积生理盐水。治疗2周后,采用苏木精-伊红(HE)染色实验和α-SMA免疫荧光染色实验评估大鼠肺血管形态;Western blot实验评估大鼠肺组织中磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(PKB/Akt)、内皮型一氧化氮合酶(eNOS)的磷酸化水平;ELISA法测定eNOS、NO水平。结果 HE染色实验和α-SMA免疫荧光染色证实丹参酮ⅡA能有效抑制PAH大鼠肺血管中膜厚度和血管肌化水平(P<0.01);Western blot实验结果显示丹参酮ⅡA能够显著提高PAH大鼠肺组织中PI3K、Akt和eNOS蛋白的磷酸化水平;ELISA实验结果显示模型组eNOS、NO水平降低(P<0.01)。结论 丹参酮ⅡA能够通过介导PI3K/Akt-eNOS信号通路改善野百合碱所致大鼠肺动脉高压。Objective To explore the therapeutic mechanism of tanshinone ⅡA in the treatment of pulmonary arterial hypertension(PAH) in rats. Methods A total of 100 male SD rats were randomized into 5 groups(n=20), and except for those in the control group with saline injection, all the rats were injected with monocrotaline(MCT) on the back of the neck to establish models of pulmonary hypertension. Two weeks after the injection, the rat models received intraperitoneal injections of tanshinone ⅡA(10 mg/kg), phosphatidylinositol 3 kinase(PI3K) inhibitor(1 mg/kg), both tanshinone ⅡA and PI3K inhibitor, or saline(model group) on a daily basis. After 2 weeks of treatment, HE staining and α-SMA immunofluorescence staining were used to evaluate the morphology of the pulmonary vessels of the rats. The phosphorylation levels of PI3K, protein kinase B(PKB/Akt) and endothelial nitric oxide synthase(eNOS) in the lung tissue were determined with Western blotting;the levels of eNOS and NO were measured using enzyme-linked immunosorbent assay(ELISA). Results The results of HE staining and α-SMA immunofluorescence staining showed that tanshinone ⅡA effectively inhibited MCT-induced pulmonary artery intimamedia thickening and muscularization of the pulmonary arterioles(P<0.01). The results of Western blotting showed that treatment with tanshinone ⅡA significantly increased the phosphorylation levels of PI3K, Akt and eNOS proteins in the lung tissue of PAH rats;ELISA results showed that the levels of eNOS and NO were significantly decreased in the rat models after tanshinone ⅡA treatment(P<0.01). Conclusion Treatment with tanshinone ⅡA can improve MCT-induced pulmonary hypertension in rats through the PI3K/Akt-eNOS signaling pathway.

关 键 词：肺动脉高压 PI3K/Akt-eNOS 丹参酮ⅡA 野百合碱 信号通路 

分 类 号：R285.5[医药卫生—中药学]