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血竭醇提物对大鼠穿支皮瓣模型存活及PI3K/Akt/eNOS通路的影响
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| 篇名： | 血竭醇提物对大鼠穿支皮瓣模型存活及PI3K/Akt/eNOS通路的影响 |
| TITLE： | |
| 摘要： | 目的：研究血竭醇提物对大鼠穿支皮瓣模型存活及PI3K/Akt/eNOS通路的影响。方法：采用只保留穿支血管切断其他血管的方法复制大鼠穿支皮瓣模型，造模成功后，将大鼠分为模型组（外敷，生理盐水）和血竭醇提取物（EESD，血竭素含量为75.08 mg/g）组（外敷，0.21 g/cm2），每组10只，连续敷药7 d，每天1次。敷药7 d后测定各组大鼠穿支皮瓣存活率、皮瓣微血管密度。将人脐静脉内皮细胞（HUVEC）缺氧缺糖16 h后复氧复糖复制HUVEC缺氧缺糖/复氧复糖细胞模型，造模成功后，将细胞分为正常组、模型组和血竭素高、中、低浓度组（2.5、1.0、0.5 μg/mL），于复氧复糖培养24 h后，采用显微镜观察各组细胞的形态，采用MTT法和比色法分别测定各组细胞的活性和细胞中一氧化氮（NO）的含量，采用逆转录-聚合酶链式反应（RT-PCR）和Western blot法检测丝氨酸/苏氨酸激酶（Akt）、磷脂酰肌醇-3-激酶（PI3K）、内皮型一氧化氮合酶（eNOS）mRNA的表达水平，PI3K蛋白的表达以及Akt、eNOS蛋白的磷酸化程度。结果：在大鼠实验中，与模型组比较，EESD组大鼠穿支皮瓣存活率、微血管密度均显著增加（P＜0.01）。在细胞试验中，与正常组比较，模型组HUVEC细胞存活率、NO含量，PI3K、Akt、eNOS mRNA的表达水平，PI3K蛋白的表达以及Akt、eNOS蛋白的磷酸化程度均显著降低（P＜0.05或P＜0.01）；与模型组比较，血竭素高、中、低浓度组HUVEC细胞的存活率、NO含量，PI3K、Akt、eNOS mRNA的表达水平，PI3K蛋白的表达以及Akt、eNOS蛋白的磷酸化程度均显著升高（P＜0.05或P＜0.01）。结论：EESD可提高大鼠穿支皮瓣模型的存活率，其机制可能与激活PI3K/Akt/eNOS通路保护内皮细胞有关。 |
| 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. |
| 期刊： | 2019年第30卷第23期 |
| 作者： | 张丽，张扬，王绪平，黄孝闻，吴人杰，寿旦 |
| AUTHORS： | ZHANG Li，ZHANG Yang，WANG Xuping，HUANG Xiaowen，WU Renjie，SHOU Dan |
| 关键字： | 血竭醇提物；血竭素；穿支皮瓣；丝氨酸/苏氨酸激酶；磷脂酰肌醇-3-激酶；内皮型一氧化氮合酶 |
| KEYWORDS： | Ethanol extract of Sanguis Draconis； Dracorhodin； Perforating flap； Akt； PI3K； eNOS |
| 阅读数： | 267 次 |
| 本月下载数： | 2 次 |
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