AUTHOR=Liu Yingqiu , Liu Tianyi , Ding Kevin , Liu Zengyuan , Li Yuanyuan , He Taotao , Zhang Weimin , Fan Yunpeng , Ma Wuren , Cui Li , Song Xiaoping TITLE=Phospholipase Cγ2 Signaling Cascade Contribute to the Antiplatelet Effect of Notoginsenoside Fc JOURNAL=Frontiers in Pharmacology VOLUME=9 YEAR=2018 URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2018.01293 DOI=10.3389/fphar.2018.01293 ISSN=1663-9812 ABSTRACT=

Scope: Bleeding, the main drawback of clinically used chemical anti-thrombotic drug is resulted from the unidirectional suppression of platelet activity. Therefore, dual-directional regulatory effect on platelet is the main preponderance of Panax notoginseng over these drugs. The dual-directional regulatory effect should be ascribed to the resourceful Panax notoginseng saponins (PNS). Clarifying the mechanism of main PNS in both inhibiting and promoting platelet aggregation will give a full outlook for the dual-directional regulatory effect. The present study is aimed at explaining the mechanism of Notoginsenoside Fc (Fc), a main PNS, in inhibiting platelet aggregation.

Methods: In the in vitro study, after incubating platelets with Fc and m-3M3FBS, platelet aggregation was triggered by thrombin, collagen or ADP. Platelet aggregation was measured by aggregometer. Phospholipase Cγ2 (PLCγ2) and protein kinase C (PKC) activities were studied by western blotting. Diacylglycerol (DAG), thromboxane B₂ (TXB₂) and 1,4,5-inositol trisphosphate (IP₃) concentrations were measured by corresponding ELISA kits. Calcium concentrations ([Ca²⁺]) were estimated through the fluorescence intensity emitted from Fluo-4. In the in vivo study, thrombus model was induced by FeCl₃. The effect of Fc on thrombosis was evaluated by measurement of protein content and observation of injured blood vessel.

Results: thrombin, collagen and ADP induced platelet aggregation were all suppressed by incubating platelets with Fc. Platelet PLCγ2 and subsequent DAG-PKC-TXA₂ and IP₃ were down-regulated by Fc as well. However, the basal [Ca²⁺] in platelet was not altered by Fc. Nevertheless, thrombin triggered activation of PLCγ2 and subsequent DAG-PKC-TXA₂ and IP₃-[Ca²⁺] were all abolished by Fc. Fc also attenuated platelet aggregation and PLCγ2 signaling activation induced by PLC activator, m-3M3FBS. In the in vivo study, FeCl₃ induced thrombosis in rat femoral artery was significantly alleviated by administration of Fc.

Conclusion: The results above suggested the antiplatelet and antithrombotic effects of Fc are carried out through oppression of PLCγ2 and subsequent DAG-PKC-TXA₂ and IP₃-[Ca²⁺]. The present study provided theoretical support for new anti-thrombotic drug exploitation by Panax notoginseng.