Tsantan Sumtang attenuated chronic hypoxia-induced right ventricular structure remodeling and fibrosis by equilibrating local ACE-AngII-AT1R/ACE2-Ang1-7-Mas axis in rat
Tsantan Sumtang attenuated chronic hypoxia-induced right ventricular structure remodeling and fibrosis by equilibrating local ACE-AngII-AT1R/ACE2-Ang1-7-Mas axis in rat
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Tsantan Sumtang 通过平衡大鼠局部 ACE-AngII-AT1R/ ACE2-Ang1-7-Mas 轴减轻慢性缺氧诱导的右心室结构重塑和纤维化
DOI：
10.1016/j.jep.2019.112470
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发表时间：
2020-03-25
影响因子：
5.4
通讯作者：
Ge, Rili
中科院分区：
文献类型：
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作者：
Dang, Zhancui;Su, Shanshan;Ge, Rili
民族药理意义：Tsantan Sumtang，由Chocropondias axillaris(Roxb.)丹参藏药L，是藏药的一种传统常用方。赞丹苏糖源于四大坦陀罗，具有滋补心脏的功效，在青海、西藏、内蒙古等地被用作治疗心脑血管疾病和心力衰竭的民间药物。本研究的目的：为阐明其减轻慢性缺氧性肺动脉高压(HPAH)大鼠右室重构和纤维化的可能机制。材料与方法：雄性SD大鼠50只，体重170+/-20g，随机分为对照组、缺氧组和缺氧+三七汤组(1.0g·kg~(-1)·d~(-1))，每组10只。1.25g·kg(-1)天(-1)1.5g·kg(-1)天(-1))。低氧组、低氧+三坛苏糖组大鼠通过调节内压和含氧量模拟海拔4500m，在低压舱中维持28d。测定平均肺动脉压(MPAP)、右室肥厚指数(RVHI)、右室重量/胫骨长度比(RV/TL)、心率(HR)和右室收缩压(RVSP)。苏木精-伊红(HE)染色观察RV结构的组织形态变化。用胶原比例面积(CPA)、I型胶原、III型胶原和羟脯氨酸含量评价RV组织纤维化程度。天狼星红染色(PSR)获得CPA。免疫组织化学和免疫印迹法检测I型和III型胶原的表达。碱性水解法测定羟脯氨酸含量。采用酶联免疫吸附试验(EL ISA)检测RV组织中血管紧张素II(AngII)和血管紧张素1-7(Ang1-7)的水平。免疫组织化学和免疫印迹法检测血管紧张素转换酶(ACE)、血管紧张素转换酶(AngII)、血管紧张素Ⅱ1型受体(AT1R)、血管紧张素转换酶2(ACE2)、Mas受体(Mas)的蛋白表达。定量聚合酶链式反应检测血管紧张素转换酶(ACE)、血管紧张素Ⅱ受体(AT1R)、血管紧张素转换酶2(ACE2)、血管紧张素转换酶(Mas)的mRNA水平。结果：HPAH大鼠的RVHI、RV/TL和RVSP均显著增加。此外，缺氧条件下RV组织中I型胶原、III型胶原和羟脯氨酸水平均上调。我们发现，血管紧张素转换酶(ACE)、血管紧张素转换酶II(AngII)、血管紧张素Ⅱ受体(AT1R)表达增加，血管紧张素转换酶2(ACE2)、血管紧张素转换酶1-7(Ang1-7)、血管紧张素转换酶(Mas)表达降低，与RV肥大和纤维化有关。Tsantan Sumtang可通过上调RV组织中ACE2、Mas水平，下调ACE、AngII、AT1R水平，减轻RV重塑和纤维化。结论：前丹苏糖具有减轻慢性缺氧大鼠右室重构和纤维化的作用。前丹苏糖的药理作用是通过平衡血管紧张素转换酶(ACE)-血管紧张素Ⅱ-血管紧张素转换酶(AT1R)和血管紧张素转换酶-血管紧张素转换酶(ACE2-Ang1-7-Mas)轴来实现的。
Ethnopharmacological relevance: Tsantan Sumtang, which consists of Chocrospondias axillaris (Roxb.) Built et Hill, Myristica fragrans Houtt and Santalum album L, is a traditional and common prescription of Tibetan medicine. Tsantan Sumtang originates from Four Tantra with properties of nourishing heart and has been used as a folk medicine for cardiovascular diseases and heart failure in Qinghai, Tibet and Inner Mongolia. Our previous studies found that Tsantan Sumtang showed beneficial effects on right ventricular structure in hypoxia rats, while the underling mechanism remains unclear.Aim of the study: To elucidate the underlying mechanisms of Tsantan Sumtang attenuated right ventricular (RV) remodeling and fibrosis of chronic hypoxia-induced pulmonary arterial hypertension (HPAH) rats.Materials and methods: Fifty male Sprague Dawley (SD) rats (170 +/- 20 g) were randomly divided into control group, hypoxia group, and hypoxia + Tsantan Sum tang groups (1.0 g.kg(-1)day(-1), 1.25 g.kg(-1)day(-1) 1.5 g.kg(-1)day(-1)). Rats in the hypoxia group and hypoxia + Tsantan Sumtang groups were maintained in a hypobaric chamber by adjusting the inner pressure and oxygen content to simulate an altitude of 4500 m for 28 days. The mean pulmonary arterial pressure (mPAP), right ventricle hypertrophy index (RVHI), the ratio of RV weight to tibia length (TL) (RV/TL), heart rate (HR) and RV systolic pressure (RVSP) was determined. Histomorphological assay of RV structure was evaluated by hematoxylin and eosin (HE) staining. RV tissue fibrosis was assessed by collagen proportion area (CPA), collagen I, collagen III and hydroxyproline content. CPA was obtained by picro-sirius red staining (PSR). The expression of collagen I and collagen III were detected by immunohistochemistry and western blotting. The hydroxyproline content was detected by alkaline hydrolysis. In addition, the level of angiotensin II (AngII) and angiotensin 1-7 (Ang1-7) in RV tissue was tested by enzyme-linked immune sorbent assay (ELISA). Protein expression of angiotensin-converting enzyme (ACE), AngII, AngII type 1 receptor (AT1R), angiotensin-converting enzyme 2 (ACE2), Mas receptor (Mas) were determined by immunohistochemistry and western blotting. mRNA level of ACE, AT1R, ACE2, Mas were tested by qPCR. The chemical profile of Tsantan Sumtang was revealed by UHPLC-Q-Exactive hybrid quadrupole-orbitrap mass analysis.Results: Our results showed that RVHI, RV/TL and RVSP were significantly increased in HPAH rat. Furthermore, levels of collagen I, collagen III and hydroxyproline were up-regulated in RV tissue under hypoxia. We found that RV hypertrophy and fibrosis were associated with increased expression of ACE, AngII, AT1R as well as decreased expression of ACE2, Ang1-7 and Mas. RV remodeling and fibrosis were attenuated after Tsantan Sumtang administration by up-regulating ACE2 and Mas level as well as down-regulating ACE, AngII and AT1R levels in RV tissue. 35 constituents in Tsantan Sumtang were identified.Conclusion: Tsantan Sumtang attenuated RV remodeling and fibrosis in rat exposed to chronic hypoxia. The pharmacological effect of Tsantan Sumtang was based on equilibrating ACE-AngII-AT1R and ACE2-Ang1-7-Mas axis of RV tissue in HPAH rat.