DFMG reverses proliferation and migration of vascular smooth muscle cells induced by co-culture with injured vascular endothelial cells via suppression of the TLR4-mediated signaling pathway

Abstract

7-Difluoromethoxy-5,4′-dimethoxy-genistein (DFMG) is a novel chemical compound synthesized using genistein. Previous studies have indicated that DFMG can reverse the apoptosis of vascular endothelial cells (VECs) by regulating the mitochondrial apoptosis pathway. The present study aimed to investigate the activity and molecular mechanism underlying DFMG-mediated protection of vascular smooth muscle cell (VSMCs) using a non-contact co-culture model established by using Transwell insert. Secretion of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by ELISA. Proliferation and migration of VSMCs were assessed using a Cell Counting kit-8 and wound healing assays, respectively. Toll-like receptor 4 (TLR4) mRNA and protein levels were detected by reverse transcription-quantitative polymerase chain reaction and western blotting analyses, respectively. In the present study, lysophosphatidylcholine (LPC) significantly increased the secretion of IL-6 and TNF-α in VECs. VECs treated with LPC markedly increased proliferation and migration of VSMCs, which were inhibited by DFMG. Transfection of either TLR4 short hairpin RNA (shRNA) or TLR4 cDNA in VECs inhibited and increased proliferation and migration of VSMCs, respectively. Furthermore, transfection of VECs with TLR4 shRNA suppressed the proliferation and migration of VSMCs induced by co-culture with injured VECs, which was further enhanced by treatment with DFMG. By contrast, transfection of VECs with TLR4 cDNA enhanced proliferation and migration of VSMCs and this effect was inhibited by treatment with DFMG. Taken together, the results of the present study demonstrated that DFMG can reverse proliferation and migration of VSMCs induced by co-culture with injured VECs via suppression of the TLR4-mediated signaling pathway.

Metabolomic analysis coupled with extreme phenotype sampling identified that lysophosphatidylcholines are associated with multisite musculoskeletal pain

“… 56 , 61 In addition, it has been reported that lysoPC can promote the expression of IL-1β and secretion of IL-6 and TNF-α in endothelial cells in vitro. 8 , 36 However, we observed a negative correlation between IL-6 and lysoPC 26:0, contradicting what was observed in those in vitro studies. 8 , 37 In vivo studies are needed to verify the role of lysoPCs in regulating cytokine expression.…”

“… 8 , 36 However, we observed a negative correlation between IL-6 and lysoPC 26:0, contradicting what was observed in those in vitro studies. 8 , 37 In vivo studies are needed to verify the role of lysoPCs in regulating cytokine expression.…”

miR-27a regulates vascular remodeling by targeting endothelial cells' apoptosis and interaction with vascular smooth muscle cells in aortic dissection

“…ECs dysfunction promotes AD development through pro-inflammatory effect 14. VSMC dysfunction can be stimulated by endothelial injury signals, and participates in the pathogenesis of vascular diseases 49. This study revealed that EC apoptosis was essential for AD in response to suppression of miR-27a.…”

The Impact of Natural Antioxidants on the Regenerative Potential of Vascular Cells

“…There is limited literature available on flavonoids and their effects on vascular regeneration; however, one of the isoflavone present in legumes, genistein, can eliminate free radicals, and boost antioxidant enzyme activities when combined with 7-difluoromethoxy-5′4-dimethoxy (DFMG). This novel antioxidant compound effectively inhibits VSMCs proliferation and migration by regulating the Toll-like receptor-4 (TLR4) signaling pathway as well as reducing oxidative stress (59) and endothelial cell impairment (60). These studies suggest that genistein may possibly prevent vascular disorders by restoring vascular function and promoting vascular cell regeneration.…”