摘要
目的:评价血红素氧合酶-1(HO-1)在七氟烷改善小鼠脓毒症相关性脑病中的作用。方法:成年雄性C57BL/6J小鼠136只,体重20~25 g,6~8周龄,采用随机数字表法分为4组(n=34):假手术组(Sham组)、脓毒症相关性脑病组(SAE组)、脓毒症相关性脑病+七氟烷组(SAE+Sevo组)和脓毒症相关性脑病+七氟烷+HO-1抑制剂锌原卟啉Ⅸ组(SAE+Sevo+ZnPPⅨ组)。采用盲肠结扎穿孔法制备小鼠脓毒症相关性脑病模型。SAE+Sevo组和SAE+Sevo+ZnPPⅨ组分别于造模成功后立即吸入2%七氟烷-33%氧气2 h,Sham组和SAE组吸入33%氧气2 h。SAE+Sevo+ZnPPⅨ组于造模前30 min时腹腔注射HO-1抑制剂ZnPPⅨ25 mg/kg。于造模后6、12和24 h时处死6只小鼠,采用ELISA法检测皮层组织TNF-α、IL-1β、高迁移率族蛋白1(HMGB1)和HO-1水平,采用Western blot法检测皮层组织HO-1表达。于造模后24 h时处死6只小鼠,采用TUNEL染色检测皮层细胞凋亡情况,计算细胞凋亡指数。每组取10只小鼠,分别于造模后3、5、7和14 d时行Y迷宫实验,计算自发交替百分比。结果:与Sham组比较,SAE组、SAE+Sevo组及SAE+Sevo+ZnPPⅨ组皮层组织TNF-α、IL-1β、HMGB1水平、细胞凋亡指数、HO-1活性及其表达水平升高,自发交替百分比降低(P<0.05)。与SAE组比较,SAE+Sevo组皮层组织TNF-α、IL-1β、HMGB1水平和细胞凋亡指数降低,HO-1活性及其表达水平、自发交替百分比升高,SAE+Sevo+ZnPPⅨ组皮层组织TNF-α、IL-1β及HMGB1水平降低(P<0.05),其余指标差异无统计学意义(P>0.05)。与SAE+Sevo组比较,SAE+Sevo+ZnPPⅨ组皮层组织TNF-α、IL-1β、HMGB1水平和细胞凋亡指数升高,HO-1活性及其表达水平、自发交替百分比降低(P<0.05)。结论:七氟烷改善小鼠脓毒症相关性脑病的机制与提高皮层组织HO-1活性,减轻炎症反应有关。
Objective To evaluate the role of heme oxygenase-1(HO-1)in sevoflurane-induced improvement in sepsis-associated encephalopathy(SAE)in mice.Methods A total of 136 adult male mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups(n=34 each)using a random number table method:sham operation group(group Sham),SAE group,SAE+sevoflurane group(group SAE+Sevo)and SAE+sevoflurane+HO-1 inhibitor Zn ProtoporphyrinⅨ(ZnPPⅨ)group(group SAE+Sevo+ZnPPⅨ).The model of SAE was established by cecal ligation and puncture(SAE)in anesthetized mice.In SAE+Sevo and SAE+Sevo+ZnPPⅨgroups,2%sevoflurane-33%oxygen was inhaled for 2 h starting from the time point immediately after establishment of the model,while 33%oxygen was inhaled for 2 h in Sham and SAE groups.ZnPPⅨ25 mg/kg was intraperitoneally injected at 30 min before the model was established in group SAE+Sevo+ZnPPⅨ.Six mice were sacrificed at 6,12 and 24 h after establishment of the model for determination of levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),high mobility group protein B1(HMGB1)and HO-1 in cortical tissues(by enzyme-linked immunosorbent assay)and the expression of HO-1(by Western blot).Another 6 mice were sacrificed for determination of apoptosis in cortical tissue(by TUNEL staining),and apoptotic index(AI)was calcultated.Ten mice in each group were selected,Y maze test was performed at 3,5,7 and 14 days after establishment of the model,and the percentage of spontaneous alternation was calculated.Results Compared with Sham group,the levels of TNF-α,IL-1βand HMGB1,AI,HO-1 activity and its expression level in cortex were significantly increased,and the percentage of spontaneous alternation was decreased in SAE,SAE+Sevo and SAE+Sevo+ZnPPⅨgroups(P<0.05).Compared with group SAE,the levels of TNF-α,IL-1βand HMGB1 and AI were significantly decreased,and HO-1 activity and its expression level and the percentage of spontaneous alternation were increased in group SAE+Sevo,the levels of TNF-α,IL-1βand HMGB1 in cortex were decreased(P
作者
王亚欣
刘玲玲
谢克亮
陈君
Wang Yaxin;Liu Lingling;Xie Keliang;Chen Jun(Department of Anesthesiology,Tianjin Huanhu Hospital,Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases,Tianjin 300350,China;Department of Anesthesiology,Tianjin Medical University General Hospital,Tianjin Research Institute of Anesthesiology,Tianjin 300052,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2021年第5期616-620,共5页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(81772043)。
