ABSTRACT
OBJECTIVES@#To investigate the role and mechanism of circRNA-SR-related CTD associated factor 8 (SCAF8) in regulating endothelial cell pyroptosis in high glucose environment.@*METHODS@#Human umbilical vein endothelial cells (HUVECs) were cultured and divided into six groups. The normal control group and high glucose control group were cultured in cell culture medium with 5 and 33 mmol/L glucose, respectively. The RNA control group, circRNA-SCAF8 inhibition group, miR-93-5p overexpression group and miR-93-5p inhibition group were added with non-functional siRNA, circRNA-SCAF8 inhibitor, miR-93-5p overexpression molecule and miR-93-5p inhibitor in high glucose environment, respectively. Cell viability and pyroptosis were detected by cell counting kit-8 (CCK-8) assay, flow cytometry and Hoechst 33342/propidium iodide fluorescence double staining. Western blotting and enzyme-linked immunosorbent assay were used to detect the expression of pyroptosis-related factors including apoptosis-associated speck-like protein containing a CARD (ASC), cysteine aspartic acid specific protease-1 (caspase-1) and Gasdermin D (GSDMD), NOD like receptor protein 3 (NLRP-3), thioredoxin interacting proteins (TXNIP), IL-18 and IL-1β. The expression of circRNA-SCAF8, miR-93-5p and TXNIP was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Fluorescence in situ hybridization (FISH) was used to locate circRNA-SCAF8 and miR-93-5p. Dual luciferase assay was used to verify the targeted regulatory relationship between miR-93-5p and upstream and downstream molecules.@*RESULTS@#Compared with the RNA control group, the cell survival rate of circRNA-SCAF8 inhibition group and miR-93-5p overexpression group increased (both P<0.01), the pyroptosis decreased (both P<0.01), and the expressions of pyroptosis-related factors such as TXNIP, NLRP-3, caspase-1, GSDMD, ASC, IL-18 and IL-1β were significantly decreased (all P<0.05). The expression of miR-93-5p was significantly increased after inhibition of circRNA-SCAF8 (P<0.01), and the expression of circRNA-SCAF8 tended to decrease after overexpression of miR-93-5p, but with no statistical significance (P>0.05). Dual luciferase assay showed that miR-93-5p downre-gulated circRNA-SCAF8 expression by binding to the 3 ´ UTR region of circRNA-SCAF8, and miR-93-5p downregulated TXNIP expression by binding to the 3 ´ UTR region of TXNIP. FISH showed that circRNA-SCAF8 and miR-93-5p were both located in the cytoplasm and were highly associated in the cells. qRT-PCR showed that the relative expression of TXNIP increased or decreased after overexpression or inhibition of miR-93-5p compared with the RNA control group, respectively (both P<0.05), suggesting that miR-93-5p could regulate TXNIP gene expression.@*CONCLUSIONS@#CircRNA-SCAF8/miR-93-5p/TXNIP axis is involved in the regulation of pyroptosis in HUVECs under high glucose.
Subject(s)
Humans , Factor VIII , RNA, Circular , Endothelial Cells , Interleukin-18 , Pyroptosis , In Situ Hybridization, Fluorescence , Caspase 1 , MicroRNAs/genetics , Carrier Proteins/genetics , RNA-Binding ProteinsABSTRACT
Objective To investigate the clinical significance of compensatory angiogenesis and the angiosome-based revascularization strategy for endovascular procedures in treating infrapopliteal critical limb ischemia in elderly diabetic patients.Methods A retrospective study of 194 patients with diabetic feet (Fontaine Ⅳ) who had undergone endovascular procedures at our hospital from January 2005 to December 2014 was performed.Based on the angiosome concept,all subjects were classified into the direct revascularization group (DR group),the indirect revascularization through collaterals group (IR-tc group) and the indirect revascularization without collaterals group (IR-wc group).After one-year follow-up,the therapeutic effects were analyzed and compared between the three groups.Results During the one-year follow-up,the rate of unhealed ulcers,the re-intervention rate and the rate of major adverse limb events (MALE) were higher in the IR-wc group than in the DR and IR-tc groups (P=0.000 and 0.000,P=0.025 and 0.013,P=0.014 and 0.023,respectively).The IR-wc group presented a lower limb salvage rate as compared with those in the DR and IR-tc groups (P=0.011 and 0.027).Patients with a single recanalized branch had a higher rate of unhealed ulcers than those with multiple recanalized branches in the IR-wc group (93.9％ vs.71.0％,P=0.015).Conclusions The angiosome-based revascularization strategy has shown important value in guiding endovascular procedures for patients with diabetic foot disease and,in particular,the compensatory angiogenesis approach appears to be critical in generating favorable short-term clinical outcomes in the treatment of diabetic foot disease.
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Objective To study the treatment effect of multilevel atherosclerotic occlusive disease of lower extremity. Methods From July 2004 to January 2008,intraoperative iliac balloon angioplasty and stenting combined with blood vassel prosthesis or autogenous reversed great saphenous vein bypass were performed on 32 patients suffering from lower extremity multilevel atheresclerotic occlusive disease. Results Surgical procedures were technically successful in all patients. The effect was good,intermittent claudication disappear, and rest pain improved. Preoperative vs postoperative ABI was 0.28±0.14 vs 0.65±0.18 (P<0.05 ).Thirty patients were followed up,the mean following period was 18 months (range of 3-36 months).Conclusions Simultaneous intravaseular interventional therapy combined with vascular bypass are effective in the treatment for patients with severe and multilevel atheroselerotie occlusive disease of lower extremity, the operation is less traumatic and the procedures are easy to do.The result is satisfactory.