Background. Deep vein thrombosis (DVT) is the third most prevalent vascular disease worldwide. MicroRNAs (miRNAs) play regulatory roles in functions of endothelial progenitor cells (EPCs), which is becoming a promising therapeutic choice for thrombus resolution. Nevertheless, the role of miR-206 in EPCs is unclear. Methods. EPCs were isolated from the peripheral blood of patients with DVT. In DVT mouse models, DVT was induced by stenosis of the inferior vena cava (IVC). The levels of miR-206 and gap junction protein alpha 1 (GJA1) in EPCs and vascular tissues of DVT mice were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The proliferation, migration, apoptosis, and angiogenesis were tested by cell counting kit-8 (CCK-8) assay, Transwell assay, flow cytometry analysis, and in vitro tube formation assay. The levels of autophagy-related proteins as well as the level of GJA1 in EPCs and vascular tissues were evaluated by western blotting. DVT formation in vivo was observed through hematoxylin-eosin (HE) staining. The expression of thrombus resolution markers, CD34 molecule (CD34) and matrix metallopeptidase 2 (MMP2), in the thrombi was measured by immunofluorescence staining. Results. miR-206 overexpression inhibited proliferation, migration, and angiogenesis and promoted apoptosis of EPCs, while miR-206 knockdown exerted an opposite effect on EPC phenotypes. Downregulation of GJA1, the target of miR-206, abolished the influence of miR-206 on EPC phenotypes. Furthermore, silencing of miR-206 suppressed the autophagy of EPCs via upregulating GJA1. miR-206 knockdown repressed thrombus formation, enhanced the homing ability of EPCs to the thrombosis site, and facilitated thrombus resolution in DVT mouse models. Additionally, miR-206 was upregulated while GJA1 was downregulated in vascular tissues of DVT mice. miR-206 knockdown elevated GJA1 expression in vascular tissues of DVT mice. The expression of miR-206 was negatively correlated with that of GJA1 in DVT mice. Conclusion. miR-206 knockdown upregulates GJA1 to inhibit autophagy of EPCs and then promote EPC proliferation, migration, and angiogenesis, thereby enhancing EPC homing to thrombi and facilitating thrombus resolution.

The purpose of this research was to evaluate HSYA' effects and mechanisms to improve Scars Induced by Anticoagulant Injection by vivo and vitro study. New Zealand rabbits were divided into Normal control (NC), Anticoagulant, Anticoagulant+HSYA-Low, Anticoagulant+HSYA-Middle and Anticoagulant+HSYA-High. Measuring TGF-β1 and IL-1β concentration by Elisa assay; evaluating pathology and fibrosis level by HE and Masson staining, measuring collage I, collage III, TLR4 and NF-κB(p65) protein expression by IHC assay. Relative gene expression (Collage I, Collage III, TLR4 and NF-κB(p65)) were evaluated by RT-qPCR assay. Relative proteins expression (Collage I, Collage III, TLR4 and NF-κB(p65)) were evaluated by WB assay. And using TGF-β1 to stimulate cell to make cell model. Compared with NC group, TGF-β1 and IL-1β concentration were significantly increased (P < 0.001); The pathology and fibrosis level were significantly deteriorated, meanwhile, Collage I, Collage III, TLR4 and NF-κB(p65) proteins and gene expression were significantly up-regulation in Anticoagulant group (P < 0.001). With HSYA supplement, TGF-β1 and IL-1β concentration were significantly depressed, Pathology and fibrosis levels were significantly improved, Collage I, Collage III, TLR4 and NF-κB(p65) proteins and gene expressions were significantly improved with dosedependent (P < 0.05). HSYA could improve anticoagulant injury induced subcutaneous scar via regulation TLR4/NF-κB(p65).

Aim: This research was to investigate the effects and mechanisms of HSYA in vascular endothelial injury by vitro study. Methods: Dividing HUVECs as Normal Control (NC), Model (LPS treated) group, HSYA-L, HSYA-M and HSYA-H groups. Cells in the HSYA treatment groups were treated with LPS, followed by 40 mg/ml, 80 mg/ml, and 120 mg/ml HSYA intervention (HSYA-L, HSYA-M, and HSYA -H groups), respectively. Measuring the cell proliferation, apoptosis, relative proteins and mRNA (TLR4, MyD88 and NF-κB(p65)) expressions by MTT, Flow cytometry, WB and RT-qPCR assay. Using cellular immunofluorescence to evaluate NF-κB(p65) nuclear volume of difference groups. Results: With HSYA supplement, the cell proliferation rates were significantly up-regulation with cell apoptosis significantly down-regulation with TLR4 relatived mRNA and proteins and NF-κB(p65) nuclear significantly depressed with dose-dependent (P <0.05, respectively). Conclusion: HSYA improved vascular endothelial injury induced by LPS via TLR4 pathway In Vitro.