泻火化瘀通窍法对感音性耳聋大鼠HMGB1/RAGE信号通路的影响  

Effects of reducing fire and resolving stasis to dredge the orific on HMGB1/RAGE signaling pathway in sensorineural deafness rats

作　　者：徐锦[1] 杨悦[1] 邹婷 熊高云[1] XU Jin;YANG Yue;ZOU Ting;XIONG Gaoyun(Department of Otolaryngology,Tongde Hospital of Zhejiang Province,Hangzhou 310013,China)

机构地区：[1]浙江省立同德医院耳鼻喉科,浙江杭州310013

出　　处：《中国现代医生》2022年第21期17-22,共6页China Modern Doctor

基　　金：浙江省中医药科技计划项目(2018ZT002)。

摘　　要：目的探讨泻火化瘀通窍法对高迁移率族蛋白B1(highy mobility group proteinboxl,HMGB1)/晚期糖基化终末产物受体(advanced glycation end product receptor,RAGE)介导的耳蜗缺血再灌注损伤(cochlea ischemia reperfusion injury,CIRI)大鼠的作用效果及机制。方法36只健康SPF级SD大鼠,雌雄各半,平均体质量(220±20)g。采用随机数字表法将大鼠分为假手术组、模型组、泻火化瘀通窍低剂量组(12.5g/kg)、泻火化瘀通窍中剂量组(25.0g/kg)、泻火化瘀通窍高剂量组(37.5g/kg)和地塞米松组(20mg/kg),每组各6只。以血管阻断法建立CIRI大鼠模型,并给予相应药物干预。测定大鼠脑干听觉诱发电位(brainstem auditory evoked potential,BAEP)阈值,苏木精-伊红染色观察大鼠耳蜗组织病变并定量评分,酶联免疫吸附法测定耳蜗组织丙二醛(malondialdehyde,MDA)、过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)的含量及活性,蛋白质印迹法检测大鼠耳蜗组织HMGB1、RAGE、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、磷酸化核因子-κB p65(phosphorylation nuclear factor-κB p65,p-NF-κB p65)、胱天蛋白酶-3(caspase-3)及活化caspase-3(cleaved-caspase-3)的蛋白表达情况。结果与假手术组相比,模型组大鼠耳蜗毛细胞变形、缺损严重,苏木精-伊红半定量评分均显著增加(P<0.01),SOD、CAT活性含量均显著降低(P<0.01),而BAEP阈值、MDA含量均显著升高(P<0.01),HMGB1、RAGE、TNF-α、IL-1β、p-NF-κB p65、cleaved-caspase-3/caspase-3蛋白表达均显著增加(P<0.01);与模型组相比,泻火化瘀通窍各剂量组和地塞米松组大鼠的CAT活性含量均显著上升(P<0.01),泻火化瘀通窍中、高剂量组和地塞米松组大鼠的苏木精-伊红半定量评分、BAEP阈值、MDA含量、HMGB1和cleaved-caspase-3/caspase-3蛋白表达均显著降低(P<0.05),泻火化瘀通窍高剂量组和地塞米松组大鼠的RAGE、p-NF-κB p65Objective To explore the effects and mechanism ofhighy mobility group proteinbox1(HMGB1)/advanced glycation end product receptor(RAGE)-mediated effect of reducing fire and resolving stasis to dredge the orifice(RRDO)on cochlea ischemia reperfusion injury(CIRI)rats.Methods Thirty-six healthy SPF SD rats,half male and half female,with an average body weight of(220±20)g.Rats were divided into sham operation group,model group,low-dose RRDO group(12.5g/kg),medium-dose RRDO group(25.0g/kg),high-dose RRDO groups(37.5g/kg)and dexamethasone group(20mg/kg)by random number table method,6 rats in each group.Vascular occlusion method was used to establish CIRI rat model,and rats were treated with the aforementioned drugs.Brainstem auditory evoked potential(BAEP)threshold of the rats was measured,hematoxylin-eosin staining was used to observe the cochlea tissue lesions and quantitative scoring,enzyme-linked immunosorbent assaywas used to detect the content and activi-ty of malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)in cochlea tissue;Western blotting method was used to detect protein of HMGB1,RAGE,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),phosphorylation nuclear factor-κB p65(p-NF-κB p65),caspase-3 and cleaved-caspase-3 expression in cochlea tis-sue.Results Compared with the sham operation group,the cochlea hair cells were deformed and severely damaged,the hematoxylin-eosin staining semi-quantitative score significantly increased(P<0.01),the activities of SOD and CAT activity significantly decreased(P<0.01),the BAEP threshold value and MDA level significantly increased(P<0.01),and the expressions of HMGB1,RAGE,TNF-α,IL-1β,p-NF-κB p65,cleaved-caspase-3/caspase-3 pro-tein significantly increased(P<0.01)in the model group.Compared with the model group,the activity of CAT in RRDO groups and dexamethasone group significantly increased(P<0.01);the BAEP threshold value,MDA level,ex-pressions of HMGB1 and cleaved-caspase-3/caspase-3 protein significantly decreased in medium,high-dose RRDO groups

关 键 词：耳蜗缺血再灌注损伤 泻火化瘀通窍法 HMGB1/RAGE 氧化应激 炎症因子 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]