Zhilong Huoxue Tongyu Capsule Alleviated the Pyroptosis of Vascular Endothelial Cells Induced by ox-LDL through miR-30b-5p/NLRP3.

Mengnan Liu,Xiaoling Lai,Raoqiong Wang,Gang Luo,Wei Ren,Tianzhu Liu,Hua Zhou,Tingfu Yang,Ping Liu,Sijin Yang,Daniel Dias Rufino Arcanjo

doi:10.1155/2022/3981350

Abstract

Background Our previous studies have demonstrated a protective role of Zhilong Huoxue Tongyu capsule in atherosclerosis (AS); however, the molecular mechanisms are unclear. Methods Human coronary artery endothelial cells (HCAECs) were induced with oxidized low-density lipoprotein (ox-LDL) to obtain cellular AS models. Then, the medicated serum of Zhilong Huoxue Tongyu capsule was obtained and used for treatment with ox-LDL-induced HCAECs. The cell viability was detected by CCK-8 assay. Besides, the binding between miR-30b-5p and NLRP3 was determined by the dual-luciferase reporter gene system assay. Furthermore, ox-LDL-induced HCAECs were transfected with miR-30b-5p mimic or miR-30b-5p inhibitor. The pyroptosis of HCAECs was assessed by flow cytometry, LDH content detection, and qRT-PCR assays. Results 10% medicated serum of Zhilong Huoxue Tongyu capsule was the maximum nontoxic concentration and it was used in subsequent assays. The rate of pyroptosis, LDH content, and the mRNA expression level of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were prominently enhanced after HCAECs were induced by ox-LDL, which were markedly rescued with medicated serum of Zhilong Huoxue Tongyu capsule. In addition, the medicated serum of Zhilong Huoxue Tongyu capsule significantly enhanced the ox-LDL-induced reduction of miR-30b-5p level. NLRP3 could bind to miR-30b-5p and was negatively corrected with miR-30b-5p. Moreover, all the rates of pyroptosis, LDH content, and the mRNA expression levels of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were further observably decreased after ox-LDL-induced HCAECs treated with medicated serum were transfected with miR-30b-5p mimic, while these were significantly rescued with transfection of miR-30b-5p inhibitor. Conclusion Zhilong Huoxue Tongyu capsule alleviated the pyroptosis of vascular endothelial cells induced by ox-LDL through miR-30b-5p/NLRP3.

Highlights

Atherosclerosis (AS) is the main cause of a variety of cardiovascular and cerebrovascular diseases, including coronary heart disease and stroke, which has brought about huge medical and economic burdens to society and individuals [1]. e occurrence and development of AS involve multiple factors
We report that Zhilong Huoxue Tongyu capsule can improve the metabolism of serum lipids and reduce the formation of carotid atherosclerotic plaques in hyperlipidemia and carotid atherosclerosis rabbits, which may be associated with the inhibition of NLRP3 inflammasome activation and downstream inflammatory factors release by suppressing the phosphorylation of NF-κB. us, we concluded that Zhilong Huoxue Tongyu capsule exhibited the antiatherosclerosis effect through NLRP3 inflammasome signaling pathway [20]
QRTPCR results exhibited that all the mRNA expression levels of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were significantly increased after Human coronary artery endothelial cells (HCAECs) were induced by oxidized low-density lipoprotein (ox-LDL), which was markedly reversed with medicated serum of Zhilong Huoxue Tongyu capsule, not blank serum (Figure 2(d)). erefore, these results indicated that the medicated serum of Zhilong Huoxue Tongyu capsule relieved the pyroptosis of HCAECs induced by oxLDL in vitro

Summary

Background

Our previous studies have demonstrated a protective role of Zhilong Huoxue Tongyu capsule in atherosclerosis (AS); the molecular mechanisms are unclear. En, the medicated serum of Zhilong Huoxue Tongyu capsule was obtained and used for treatment with ox-LDL-induced HCAECs. e cell viability was detected by CCK-8 assay. E rate of pyroptosis, LDH content, and the mRNA expression level of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were prominently enhanced after HCAECs were induced by ox-LDL, which were markedly rescued with medicated serum of Zhilong Huoxue Tongyu capsule. All the rates of pyroptosis, LDH content, and the mRNA expression levels of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were further observably decreased after ox-LDL-induced HCAECs treated with medicated serum were transfected with miR-30b-5p mimic, while these were significantly rescued with transfection of miR-30b-5p inhibitor. Zhilong Huoxue Tongyu capsule alleviated the pyroptosis of vascular endothelial cells induced by ox-LDL through miR-30b-5p/NLRP3

Introduction

Materials and Methods

Results