Qingda Granule Attenuates Angiotensin II-Induced Blood Pressure and Inhibits Ca2+/ERK Signaling Pathway

Meizhu Wu; Xiangyan Wu; Ying Cheng; Zhiqing Shen; Xiaoping Chen; Qiurong Xie; Jianfeng Chu; Jiapeng Li; Liya Liu; Lihui Wei; Linzi Long; Qiaoyan Cai; Jun Peng; Aling Shen

doi:10.3389/fphar.2021.688877

Qingda Granule Attenuates Angiotensin II-Induced Blood Pressure and Inhibits Ca²⁺/ERK Signaling Pathway

Front Pharmacol. 2021 Jul 29:12:688877. doi: 10.3389/fphar.2021.688877. eCollection 2021.

Authors

Meizhu Wu^{1

2

3}, Xiangyan Wu^{1

2

3}, Ying Cheng^{1

2

3}, Zhiqing Shen^{1

2

3}, Xiaoping Chen^{1

2

3}, Qiurong Xie^{1

2

3}, Jianfeng Chu^{1

2

3}, Jiapeng Li⁴, Liya Liu^{1

2

3}, Lihui Wei^{1

2

3}, Linzi Long^{1

2

3

5}, Qiaoyan Cai^{1

2

3}, Jun Peng^{1

2

3}, Aling Shen^{1

2

3}

Affiliations

¹ Academy of Integrative Medicine, Fuzhou, China.
² Chen Keji Academic Thought Inheritance Studio, Fuzhou, China.
³ Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou, China.
⁴ Department of Physical Education, Fujian University of Traditional Chinese Medicine, Fuzhou, China.
⁵ Department of Geriatrics, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China.

Abstract

Objective: As a well-known traditional Chinese medicine formula prescribed by academician Ke-ji Chen, Qingda granule (QDG) lowered the blood pressure of spontaneously hypertensive rats and attenuated hypertensive cardiac remodeling and inflammation. However, its functional role and underlying mechanisms on hypertensive vascular function remain largely unclear. This study aims to assess the effects of QDG treatment on Angiotensin II- (AngII-) induced hypertension and vascular function and explore its underlying mechanisms both in vitro and in vivo. Methods: In an in vivo study, 25 male C57BL/6 mice were randomly divided into five groups, including Control, AngII, AngII + QDG-L, AngII + QDG-M, and AngII + QDG-H groups (n = 5 for each group). Mice in AngII and AngII + QDG-L/-M/-H groups were infused with AngII (500 ng/kg/min), while in the Control group, they were infused with saline. Mice in AngII + QDG were intragastrically given different concentrations of QDG (0.5725, 1.145, or 2.29 g/kg/day), while in Control and AngII groups, they were intragastrically given equal volumes of double distilled water for 2 weeks. Blood pressure was determined at 0, 1, and 2 weeks of treatment. Ultrasound was used to detect the pulse wave velocity (PWV) and HE staining to detect the pathological change of the abdominal aorta. RNA sequencing (RNA-seq) was performed to identify the differentially expressed transcripts (DETs) and related signaling pathways. IHC was used to detect the expression of p-ERK in the abdominal aorta. Primary isolated rat vascular smooth muscle cells (VSMCs) were used to assess the cellular Ca²⁺ release and activation of the ERK pathway by confocal microscope and western blotting analysis, respectively. Results: QDG treatment significantly alleviated the elevated blood pressure, the PWV, and the thickness of the abdominal aorta in AngII-induced hypertensive mice. RNA-seq and KEGG analyses identified 1,505 DETs and multiple enriched pathways (including vascular contraction and calcium signaling pathway) after QDG treatment. Furthermore, confocal microscope showed that QDG treatment partially attenuated the increase of Ca²⁺ release with the stimulation of AngII in cultured VSMCs. In addition, IHC and western blotting indicated that QDG treatment also partially alleviated the increase of phospho-ERK levels in abdominal aorta tissues of mice and cultured VSMCs after the infusion or stimulation of AngII. Conclusion: QDG treatment attenuated the elevation of blood pressure, abdominal aorta dysfunction, pathological changes, Ca²⁺ release, and activation of the ERK signaling pathway.

Keywords: Ca2+/ERK signaling pathway; Qingda granule; angiotensin II; hypertension; vascular smooth muscle cells.