Extract from Rostellularia procumbens (L.) Nees Inhibits Thrombosis and Platelet Aggregation by Regulating Integrin β3 and MAPK Pathways

Ying Zhang; Zongchao Hong; Zixin Yuan; Tianshun Wang; Xingpan Wu; Bo Liu; Zhongzhu Ai; Hezhen Wu; Yanfang Yang

doi:10.1021/acsomega.0c05227

Extract from Rostellularia procumbens (L.) Nees Inhibits Thrombosis and Platelet Aggregation by Regulating Integrin β₃ and MAPK Pathways

ACS Omega. 2020 Dec 3;5(49):32123-32130. doi: 10.1021/acsomega.0c05227. eCollection 2020 Dec 15.

Authors

Ying Zhang¹, Zongchao Hong¹, Zixin Yuan¹, Tianshun Wang¹, Xingpan Wu¹, Bo Liu¹, Zhongzhu Ai¹, Hezhen Wu^{1

2

3}, Yanfang Yang^{1

2

3}

Affiliations

¹ Faculty of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China.
² Key Laboratory of Traditional Chinese Medicine Resources and Chemistry of Hubei Province, Hubei University of Chinese Medicine, Wuhan 430065, China.
³ Collaborative Innovation Center of Traditional Chinese Medicine of New Products for Geriatrics Hubei Province, Hubei University of Chinese Medicine, Wuhan 430065, China.

Abstract

Aim of study: The main objective of this study was to investigate the antithrombotic and antiplatelet effect of the extract from Rostellularia procumbenss (L.) Nees and understand the mechanisms by which it exerts its antithrombotic and antiplatelet mechanisms.

Materials and methods: The antithrombotic effective parts (RPE) were isolated using D101 macroporous adsorption resin and potential active ingredients (JAC) were isolated using the preparative liquid-phase method. The lactate dehydrogenase kit was used to determine the toxicity of RPE and JAC to platelets. The antiadhesion effect of RPE and JAC on platelets was observed by fluorescence microscopy with rhodamine phalloidin. Antithrombotic efficacy of RPE and JAC in vivo was evaluated by establishing a rat tail thrombosis model. Contents of p-selectin, TXB₂, and 6-keto-PGF_1α in rat serum were measured using an enzyme-linked immunosorbent (ELISA) assay, and the rat black tail rate was measured to prove the protective effect of RPE and JAC on the tail thrombus rat model. Western blot was used for detection of serum-related proteins in the tail thrombus rat model.

Results: The results showed that RPE had antithrombotic and antiplatelet effects. RPE and JAC have no toxicity to platelets. In vitro experiments showed that RPE and JAC had antiadhesion effects on platelets. In vivo experiments showed that RPE significantly inhibited the increase of p-selectin and TXB₂ and significantly increased the content of 6-keto-PGF_1α in the serum of rats. Western blot results demonstrated that RPE and JDB significantly inhibited the phosphorylation of the MAPK protein family in the platelets of rats, and RPE also significantly inhibited the phosphorylation of β₃ protein.

Conclusions: RPE has antithrombotic and antiplatelet activity in vivo and vitro. Its mechanism may be via preventing integrin α_IIbβ₃ activation, which in turn leads to the inhibition of the phosphorylation of the MAPK family and further suppresses TXA₂, which leads to the antithrombotic and antiplatelet effects.