黄芪甲苷通过调控TLR-4/NF-κB信号通路对大鼠皮瓣缺血再灌注损伤的影响  

Effect of Astragaloside on Ischemia-reperfusion Injuryin Rat Skin Flap by Regulating TLR-4/NF-κB Signaling Pathway

作　　者：王飞[1] 田阳 徐骁然 王德华[1] 李庭超 WANG Fei;TIAN Yang;XU Xiaoran;WANG Dehua;LI Tingchao(Department of Burns and Plastic Surgery,the First People's Hospital of Zunyi,the Third Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China;Department of Pathology,the First People's Hospital of Zunyi,the Third Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义市第一人民医院,遵义医科大学直属第三附属医院烧伤整形外科,贵州遵义563000 [2]遵义市第一人民医院,遵义医科大学直属第三附属医院病理科,贵州遵义563000

出　　处：《中国皮肤性病学杂志》2021年第5期497-503,共7页The Chinese Journal of Dermatovenereology

摘　　要：目的探讨黄芪甲苷(astragaloside,AS)对大鼠皮瓣缺血再灌注损伤的影响及其可能机制。方法按随机数字表法将SD大鼠分为假手术组(Sham)、模型组(Model)、AS低剂量组(AS-L,20 mg/kg)和AS高剂量组(AS-H,80 mg/kg),每组12只。术后立即给予药物灌胃处理,1次/d,连续7 d。观察术后2、4、6 d各组大鼠皮瓣变化情况,并于术后7 d测定各组大鼠皮瓣成活率;HE染色观察大鼠皮瓣组织病理学变化;ELISA实验检测大鼠血清炎症因子TNF-α、IL-6和IL-1β表达水平;比色法测定大鼠组织SOD活性和MDA含量;qRT-PCR检测大鼠皮瓣组织TLR4和NF-κB p65 mRNA表达水平;Western blot检测大鼠皮瓣组织全蛋白TLR4、MyD88以及核蛋白NF-κB p65表达水平。结果Model组大鼠腹部皮瓣出现严重水肿,有炎症渗出,皮瓣发硬,毛发生长不良;不同剂量AS处理后能有效缓解皮瓣缺血再灌注引起的水肿和炎性反应。与Sham组比较,Model组大鼠皮瓣存活率和皮瓣组织中SOD活性显著降低(P<0.05),而血清TNF-α、IL-6和IL-1β表达水平、皮瓣组织中MDA含量、TLR4和NF-κB p65 mRNA表达水平以及全蛋白TLR4、MyD88和核蛋白NF-κB p65表达水平均显著增加(P<0.05);与Model组比较,AS-L组和AS-H组大鼠皮瓣存活率和皮瓣组织中SOD活性均显著提高(P<0.05),而血清TNF-α、IL-6和IL-1β表达水平、皮瓣组织中MDA含量、TLR4和NF-κB p65 mRNA表达水平以及全蛋白TLR4、MyD88和核蛋白NF-κB p65表达水平均显著降低(P<0.05)。结论黄芪甲苷能改善大鼠皮瓣缺血再灌注损伤,其机制可能与抑制TLR-4/NF-κB信号通路介导的炎症反应和氧化损伤有关。Objective To investigate the effect of Astragaloside(AS)on ischemia-reperfusion injury in rat skin flap and its possible mechanism.Methods According to the random number table method,SD rats were divided into sham operation group(Sham),model group(Model),low-dose AS group(AS-L,20 mg/kg)and high-dose AS group(AS-H,80 mg/kg),with 12 rats in each group.The drug was given by intragastic administration immediately after operation,once a day for 7 consecutive days.The changes of rat skin flaps in each group at 2,4,and 6 d after operation were observed,and the survival rate of rat skin flaps in each group at 7 d after operation was measured.HE staining was used to observe the pathological changes of rat skin flaps.ELISA was used to detect the levels of serum inflammatory factors TNF-α,IL-6 and IL-1βin rats.Colorimetric method was used to detect SOD activity and MDA levels of rat tissue.The expression of rat skin flap tissue TLR4 and NF-κB p65 mRNA was measured by qRT-PCR method.The expression levels of total protein TLR4,MyD88 and nuclear protein NF-κB p65 was detected by Western blot method in rat skin flap tissue.Results The abdominal skin flap in the model group showed severe edema,inflammatory exudation,hard skin flap,and poor hair growth.Different dose of AS treatment could effectively alleviate the edema and inflammatory reaction caused by ischemia-reperfusion in skin flap.Compared with the sham group,the skin flap survival rate and the SOD activity of the skin flap tissue in the model group were significantly reduced(P<0.05),while the serum TNF-α,IL-6 and IL-1βexpression levels,the flap tissue MDA level,TLR4 and NF-κB p65 mRNA expression levels,the expression of the whole protein TLR4,MyD88 and nuclear protein NF-κB p65 were increased significantly(P<0.05).Compared with the model group,rat skin flap survival rate and flap tissue SOD activity were increased significantly(P<0.05),while serum TNF-α,IL-6 and IL-1βexpression levels,and skin flap tissue MDA level,the expression of TLR4 and NF-κB p65 mRNA and

关 键 词：黄芪甲苷 缺血再灌注损伤 皮瓣 TLR-4/NF-κB信号通路 炎症反应 氧化损伤 

分 类 号：R751[医药卫生—皮肤病学与性病学]