摘要
目的:研究活化蛋白C(Activated protein C,APC)对大鼠皮瓣缺血再灌注损伤的作用及可能机制。方法:将80只SD雄性大鼠随机分为四组:对照组、药物对照组、模型组和治疗组,每组20只。观察术后72 h内模型大鼠皮瓣形态变化,HE染色观察大鼠皮瓣组织病理学变化,TUNEL法染色观察皮瓣组织细胞凋亡,ELISA法检测血清中TNF-α、IL-6水平,用黄嘌呤氧化酶法和硫代巴比妥酸TBA比色法分别测定皮瓣组织中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,Westernblot法检测皮瓣组织中Nrf-2、HO-1、γ-GCS蛋白表达水平。结果:与模型组比较,治疗组皮瓣红肿、坏死程度、病理损伤程度减弱;TUNEL法染色观察,与模型组比较,治疗组皮瓣组织细胞凋亡率减少(P<0.05);ELISA法检测发现,与模型组比较,治疗组血清中TNF-α、IL-6降低(P<0.05);与模型组比较,治疗组SOD活性升高(P<0.05),MDA含量降低(P<0.05);Westernblot法检测发现,与模型组比较,治疗组Nrf-2、HO-1、γ-GCS蛋白相对表达水平上升(P<0.05)。结论:APC能改善大鼠皮瓣缺血再灌注损伤,其机制可能与激活Nrf-2/HO-1信号通路,抑制氧化应激反应和减少细胞凋亡、炎症反应有关。
Objective To investigate the effect of activated protein C(APC)on rat skinflap ischemia reperfusion injury and its possible mechanism.Methods Eighty male SD rats were randomly divided into 4 groups:control group,model group,treatment group and drug control group,with 20 rats in each group.The morphological changes of skinflap of model rats were observed 72 h after operation.The histopathological changes of rat skinflaps were observed by HE staining.The apoptosis of skinflap tissue was observed by TUNEL staining.The levels of TNF-αand IL-6 in serum were determined by ELISA.SOD activity and MDA content of skinflap tissue were determined by xanthine oxidase and thiobarbituric acid(TBA)colorimetry.The protein expression levels of Nrf-2,HO-1 andγ-GCS in skinflap tissues were detected by Westernblot.Results Compared with the model group,the degree of flaps redness and swelling,necrosis and pathological injury in the treatment group decreased.TUNEL staining showed that compared with the model group,the apoptosis rate in the treatment group was decreased(P<0.05).Compared with the model group,TNF-αand IL-6 in the treatment group were decreased by ELISA(P<0.05).Compared with model group,SOD activity in treatment group was increased(P<0.05),while MDA content was decreased(P<0.05).Compared with the model group,the relative expression levels of Nrf-2,HO-1 andγ-GCS were increased in the treatment group by westernblot(P<0.05).Conclusion APC can improve the rat skin flap ischemia-reperfusion injury and its mechanism may be related to the activation of Nrf-2/HO-1 signaling pathway,inhibition of oxidative stress response,reduction cell apoptosis and inflammatory response.
作者
王浩
杨建强
张彬
柯友群
湛梅圣
WANGHao;YANGJianqiang;ZHANGBin;KEYouqun;ZHANMeisheng(Department of Orthopedics,Zaoyang First People's Hospital,Zaoyang 441200,Hubei,China)
出处
《中国美容医学》
CAS
2023年第4期66-70,共5页
Chinese Journal of Aesthetic Medicine