Identification and functional characterization of a biflavone as a novel inhibitor of TRPV4‐dependent proatherogenic processes in macrophages

Abstract

Atherosclerosis, a chronic inflammatory disease, is the major cause of mortality and morbidity in the United States. Studies suggest that arterial stiffness is a marker and a risk factor for atherosclerosis. Recent reports from our laboratory showed that TRPV4 (transient receptor potential channel of the vanilloid subfamily 4), a mechanosensitive ion channel, plays a role in oxidized LDL (oxLDL)‐induced macrophage foam cell formation, a critical process in atherogenesis. We performed a high‐throughput screening for antagonists of TRPV4 from a library of 2000 natural compounds using a fluorometric imaging plate reader (FLIPR)‐based Ca2+ influx assay. We identified ginkgetin, a biflavone, as a novel small molecule chemical inhibitor of TRPV4. Primary and secondary screening was performed with mouse bone marrow derived macrophages (BMDMs) to assess the ability of ginkgetin and other candidates to inhibit TRPV4‐dependent Ca2+ influx. In our hand’s, ginkgetin demonstrated an IC50 of 0.5 μM for inhibition of TRPV4‐dependent Ca2+ influx. Furthermore, we found that antagonism of TRPV4 by ginkgetin blocks oxLDL‐induced macrophage foam cell formation. Moreover, we found that ginkgetin treatment 1) attenuated oxLDL uptake but not its cell surface binding in BMDMs, 2) inhibited lipopolysaccharide‐induced phosphorylation of JNK, and 3) inhibited oxLDL‐induced expression of proinflammatory cytokines. Taken together, our results show that ginkgetin is a novel inhibitor of TRPV4‐mediated proatherogenic processes in macrophages. Support or Funding Information This work was supported by NIH (1R01EB024556‐01) and NSF (CMMI‐1662776) grants to Shaik O. Rahaman.

Neuroprotective Potential of Biflavone Ginkgetin: A Review

“…It may also be useful for the treatment of cardiovascular disease. Cell-based studies showed its potential as an inhibitor of TRPV4-mediated proatherogenic processes in macrophages [ 50 ]. In addition, ginkgetin showed inhibitory effects on human thrombin, an important serine protease that regulates the blood coagulation cascade and processes of thrombosis [ 51 ], and pancreatic lipase, an important target that regulates lipid uptake [ 52 ].…”

“…Flavonoids are widely recognized as a molecule with good antioxidant activity and beneficial effects in the treatment of neurological disorders [ 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 ,…”