Study on the Effects of Ethanol Extract of Sanguis Draconis on the Survival of Perforating Flap Model in Rats and PI 3K/Akt/ eNOS Pathway / 中国药房

ABSTRACT

OBJECTIVE： To study the effects of ethanol extract of Sanguis Draconis on the survival of perforating flap model in rats and PI3K/Akt/eNOS pathway. METHODS： Perforating flap model was established by cutting off surrounding vessels and keeping one perforator. After modeling， the rats were divided into model group （external use， normal saline） and ethanol extract of Sanguis Draconis （EESD， the content of dracorhodin was 75.08 mg/g） group （external use， 0.21 g/cm2）， with 10 rats in each group. They were given relevant medicine for consecutive 7 days， once a day. The flap survival rate and flap microvessel density were determined after given relevant medicine 7 days. Human umbilical vein endothelial cells （HUVECs） were reoxygenated and glycoconjugated 16 h after hypoxia and hypoglycemia to establish oxygen-glucose deprivation/oxygen-glucose recovery model of HUVECs. After modeling， model cells were divided into normal group， model group， dracorhodin high-concentration， medium- concentration and high-concentration groups （2.5， 1.0， 0.5 μg/mL）. After reoxygenated and glycoconjugated for 24 h， cells morphology was observed by microscope； cell viability and the content of NO were detected by MTT assay and colorimetry. mRNA expression of Akt， PI3K and eNOS， PI3K protein expression， the phosphorylation of Akt and eNOS protein were determined by RT-PCR and Western blot assay. RESULTS： In rat experiment， compared with model group， flap survival rate and microvessel density of rats were increased significantly in EESD group （P＜0.01）. In cell experiment， compared with normal group， the survival rate of HUVEC， NO content， mRNA expression of PI3K， Akt， eNOS，PI3K protein expression， the phosphorylation of Akt and eNOS protein were decreased significantly （P＜0.05 or P＜0.01）. Compared with model group， dracorhodin high-concentration， medium-concentration and high-concentration groups survival rate of HUVEC cells， NO content， mRNA expression of PI3K， Akt and eNOS， PI3K protein expression， the phosphorylation of Akt and eNOS protein were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The survival rate of perforating flap model in rat can be increased by treating with EESD， the mechanism of which may be associated with the activation of PI3K/Akt/eNOS pathway to protect endothelial cells.