First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan 250355, China; Central Laboratory, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250014, China.
First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan 250355, China.
J Ethnopharmacol. 2021 Feb 10;266:113453. doi: 10.1016/j.jep.2020.113453. Epub 2020 Oct 8.
Banxia Baizhu Tianma decoction (BBTD) is a classical representative prescription for expelling phlegm, extinguishing wind, strengthening the spleen and dissipating excessive fluid in traditional Chinese medicine (TCM). According to both TCM theory and about 300 years of clinical practice, BBTD is especially suitable for hypertensive patients of abdominal obesity and lacking physical activity.
The present study tried to interpret the pharmacology of the ancient formula of BBTD. Herein, we focused on the plasma metabonomics of BBTD and evaluated the effect and targets of BBTD on endothelial protective effect.
Obesity-related hypertensive mice were induced by high-fat diet for 20 weeks. BBTD (17.8 g/kg) was administered intragastrically for 8 weeks, and telmisartan group (12.5 mg/kg) was used as positive drug. Body weight, blood pressure, triglyceride and cholesterol were recorded to evaluate the efficacy of BBTD in vivo. Lipid deposition in aortic roots was assessed by oil red O staining, while morphology of aortas was observed by HE staining. Ultra performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) was performed to study the plasma non-targeted metabonomics. According to the data of metabonomics, human aortic endothelial cells (HAECs) were treated by oxidized low-density lipoprotein (ox-LDL, 50 μg/mL) with/without BBTD (2, 1 or 0.5 mg/mL). Apoptosis rate (Annexin V-FITC/PI), migration (Transwell), cytoskeleton (Phalloidin) and density of VE-cadherin (Immunofluorescence staining) were used to investigate the effect of BBTD in vitro. Transcriptome sequencing was performed (2 mg/mL BBTD vs ox-LDL) to screen the possible targets of BBTD in endothelial protection against ox-LDL.
BBTD effectively reduced the body weight and total cholesterol, and decreased 12.1 mmHg in SBP and 10.5 mmHg in DBP of obesity-related hypertensive mice (P < 0.05). BBTD attenuated lipid deposition in arterial roots and improved the morphology of aortas in vivo. Plasma metabolite profiles identified 94 differential metabolites and suggested BBTD mainly affected glycerophospholipids and fatty acyls. Bioinformatics analysis indicated sphingolipid metabolism and fluid shear stress and atherosclerosis were main pathways. Therefore, we focused on endothelial protective effect of BBTD against ox-LDL. In vitro, BBTD demonstrated endothelial protective effects, decreasing apoptosis rate, improving cell migration in dose-dependent manner and maintaining cell morphology. Transcriptome sequencing identified 251 downregulated and 603 upregulated mRNAs after 24h-BBTD treatment, which reversed 51.8% change in mRNAs (393 DE mRNAs) induced by ox-LDL. Bioinformatics analysis supported the potential of BBTD in hypertension and suggested that BBTD improved endothelial cells by targeting mainly on p53 and PPAR signaling pathways.
BBTD attenuates obesity-related hypertension by regulating metabolism of glycerophospholipids and endothelial protection.
半夏白术天麻汤（BBTD）是一种经典的祛痰、熄风、健脾、消饮的中药方剂。根据中医理论和 300 多年的临床实践，BBTD 特别适用于腹部肥胖和缺乏身体活动的高血压患者。
本研究试图解释 BBTD 古代配方的药理学。在此，我们专注于 BBTD 的血浆代谢组学，并评估 BBTD 对内皮保护作用的效果和靶点。
通过高脂肪饮食诱导肥胖相关高血压小鼠 20 周。BBTD（17.8g/kg）灌胃 8 周，替米沙坦组（12.5mg/kg）作为阳性药物。记录体重、血压、甘油三酯和胆固醇，以评估 BBTD 的体内疗效。用油红 O 染色评估主动脉根部的脂质沉积，用 HE 染色观察主动脉形态。采用超高效液相色谱/串联质谱（UPLC-MS/MS）进行血浆非靶向代谢组学研究。根据代谢组学数据，用氧化型低密度脂蛋白（ox-LDL，50μg/mL）处理人主动脉内皮细胞（HAECs），并加入 BBTD（2、1 或 0.5mg/mL）。用 Annexin V-FITC/PI 检测细胞凋亡率，用 Transwell 检测细胞迁移，用鬼笔环肽检测细胞骨架，用免疫荧光染色检测 VE-钙黏蛋白密度，以研究 BBTD 的体外作用。用转录组测序（2mg/mL BBTD 与 ox-LDL）筛选 BBTD 在内皮细胞保护作用中对 ox-LDL 的可能靶点。
BBTD 有效降低肥胖相关高血压小鼠的体重和总胆固醇，分别降低收缩压 12.1mmHg 和舒张压 10.5mmHg（P<0.05）。BBTD 减轻了动脉根部的脂质沉积，改善了体内主动脉的形态。血浆代谢物谱鉴定出 94 种差异代谢物，并提示 BBTD 主要影响甘油磷脂和脂肪酸酰基。生物信息学分析表明，鞘脂代谢和血流切应力与动脉粥样硬化是主要途径。因此，我们专注于 BBTD 对 ox-LDL 的内皮保护作用。在体外，BBTD 表现出内皮保护作用，呈剂量依赖性降低细胞凋亡率，提高细胞迁移能力，维持细胞形态。转录组测序显示，ox-LDL 处理 24 小时后有 251 个下调和 603 个上调的 mRNAs，ox-LDL 诱导的 393 个差异表达 mRNAs（DE mRNAs）中有 51.8%得到逆转。生物信息学分析支持 BBTD 在高血压中的潜在作用，并表明 BBTD 通过主要针对 p53 和 PPAR 信号通路来改善内皮细胞。
BBTD 通过调节甘油磷脂代谢和内皮保护作用来减轻肥胖相关高血压。
