ABSTRACT
Objective To investigate the protective mechanism of hypoxia-inducible factor-1α (HIF-1α) in vascular endothelial cells under hypoxia.Methods 1.After a hemorrhagic shock model was established in mice,the vascular endothelial cells were sorted in a shock group (n =3) and a sham operation group (n =3) for RNA-sequencing to analyze the main differential molecules.2.The expression of HIF-1α and glucose transporter-1 (GLUT1) was measured in human umbilical vein endothelial cells (HUVECs) and the mitochondrial membrane potentials were detected in a control group (normal culture,n =3) and a hypoxia group (hypoxia culture for 6 h,n =3).3.The HUVECs cells were transfected with HIF-1α siRNA for 48 h to interfere with HIF-1 α expression,and the control group(n =3) was transfected with control siRNA.The expression of HIF-1α was detected to determine the interference effect.The mRNA and protein expression of GLUT1 was detected in the interference and the control groups after 6 h of hypoxia culture.The mitochondrial membrane potential was detected by fluorescent probe method.Results 1.The transcriptome sequencing in the vascular endothelial cells in the shock and sham operation groups indicated 25 genes with significant differences.The HIF-1 α expression was significantly increased in the shock group (111.70 ± 15.97) than in the sham operation group (53.49 ± 3.26) (P =0.023).2.The expression of HIF-1 α and GLUT1 in the HUVECs cells was significantly increased in the hypoxia group compared with the control group (P ＜ 0.05).The mitochondrial membrane potential was significantly lower in the hypoxia group (0.781 ± 0.023) than in the control group (1.177 ± 0.062) (P ＜ 0.05),indicating aggravated injury.3.There was no significant difference in the mitochondrial membrane potential between the interference group (1.011 ± 0.076) and the control group (1.151 ± 0.031) (P ＞ 0.05).However,after hypoxia culture for 6 h,there was a significant difference in the mitochondrial membrane potential between the interference group (0.514 ± 0.018) and the control group (0.769 ± 0.044) (P ＜ 0.05),also indicating aggravated damage.Conclusions The expression of HIF-1 α may be significantly increased in hemorrhagic shock.In HUVECs under hypoxia,HIF-1 α may up-regulate the expression of GLUT1,promote glucose transport,improve mitochondrial damage and protect vascular endothelial cells.Thus,targeting HIF-1 α may contribute to the treatment of hemorrhagic shock.
ABSTRACT
Objective To explore the curative effect of fibular flap with limbs composite soft-tissue.Methods From February,2013 to February,2016,13 cases with body severe trauma patients were treated,which including 5 cases of upper limbs and 8 cases of lower limbs,and all existed bone defect,soft tissue defect and trunk vessel defect.Three cases with limbs distal non blood supply were emergency treated with debridment and flow-through fibular flap transplantation renovation,peroneal artery repairing defective blood vesscls to rcstorc limbs distal blood supply,fibular flap repairing bone defect,skin flap repairing soft tissue defect.The limb blood supply for other 10 cases were in good condition,but one case with main artery defect did the second phase of fibular flap transplantation and repaired defective blood vessels,bone and skin soft tissue synchronously according to wound condition.According to the postoperative observation for flap survival and appearance,X-ray films to observe fracture healing after 6 weeks,three months and 6 months of operation as well as evaluating limb function recovery,then analyzed the results.Results Flaps survived successfully for 11 cases,and flaps for the other 2 cases were partial necrosis.One Case was edge flap necrosis,heal scabby after dressing,and the other case was necrosis for 1/3 of the area,but the deep fascia survival,and the skin graft healing after dressing.One case with forearm rolling was in vascular crisis after operation,but tbe crisis was relieved after detection,and fingers blood supply was recovered.All the patients were followed up for 6 to 36 months(mean,14 months).All flaps were survived,fractures healed well and limbs distal blood supply was good.Bone healing time was 8 to 24 weeks,and patients with lower limbs injury could bear load after 3 to 8 months.Lower limbs restored walking function.Upper limbs and hands restored rotation function.Transplant flapshad good elasticity and satisfactory appearance.Conclusion Using fibular flap to repair defective blood vessels,bone and soft tissue synchronously,not only can rescue the limbs on the verge of amputation,but also can repair defective composite tissue and get a good prognosis.It is an effective method for open injuries severely treatment in clinic.