清瘟败毒饮对脓毒症小鼠脾组织微小RNA表达的影响  

Effect of Qingwen Baidu Decoction on the expression of microRNA in spleen tissue of septic mice

作　　者：林名瑞[1] 文丹[2] 乐道平[3] 李玮[1] Lin Mingrui;Wen Dan;Le Daoping;Li Wei(Department of Intensive Care Unit,the People's Hospital of Fujian Traditional Medical University,Fuzhou 350004,Fujian,China;Department of Emergency,the People's Hospital of Fujian Traditional Medical University,Fuzhou 350004,Fujian,China;Department of Intensive Care Unit,Ningde Hospital,Ningde 352100,Fujian,China)

机构地区：[1]福建中医药大学附属人民医院重症医学科,福州350004 [2]福建中医药大学附属人民医院急诊科,福州350004 [3]宁德市医院重症医学科,福建宁德352100

出　　处：《中国中西医结合急救杂志》2023年第1期18-21,共4页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：福建省医疗“创双高”重点专科(2017-881);福建省卫生健康青年科研课题(2016-2-31)。

摘　　要：目的采用小RNA(sRNA)测序技术检测清瘟败毒饮干预的脓毒症小鼠24 h脾组织微小RNA(miRNA)的表达变化,从而从非编码基因层面分析清瘟败毒饮抗脓毒性脾功能损伤的可能机制。方法将45只SPF级健康BALB/C小鼠随机等分为对照组、脓毒症组和清瘟败毒饮组。清瘟败毒饮成分每剂含生石膏15 g、生地黄7.5 g、水牛角15 g、黄连2.5 g、栀子5 g、桔梗5 g、黄芩5 g、知母5 g、赤芍5 g、玄参5 g、连翘5 g、甘草5 g、丹皮5 g、竹叶5 g。脓毒症组和清瘟败毒饮组小鼠腹腔注射脂多糖(LPS)10 mg/kg构建脓毒症模型,对照组腹腔注射等量生理盐水。清瘟败毒饮组于制模前2 d开始给予清瘟败毒饮药液0.3 mL,每1 mL相当于生药1.5 g灌胃,术后继续胃饲,每日2次;对照组及脓毒症组给予等量生理盐水胃饲。术后24 h各组随机取5只小鼠脾组织,每组各取1只小鼠的脾组织于光镜下观察病理学改变,提取其余小鼠脾组织总RNA,采用sRNA测序技术进行miRNA检测,应用生物信息学软件分析脓毒症小鼠脾组织miRNA表达差异及靶基因的预测。结果光镜下显示,对照组小鼠脾组织结构正常;脓毒症组小鼠脾组织白髓总体结构稍紊乱,红白髓交界偏模糊,红髓充血明显;清瘟败毒饮组小鼠脾组织紊乱结构较前有所改善。与对照组比较,脓毒症组小鼠脾组织已知miRNA表达上调数为39个,下调数为16个;清瘟败毒饮组小鼠脾组织已知miRNA表达上调数为6个,下调数为24个。经LPS干预后,miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p表达明显上调;经清瘟败毒饮干预后,miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p表达出现一定程度下调。结论清瘟败毒饮可改善脓毒症小鼠脾组织结构改变,可能通过下调miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p基因表达起到保护脓毒症小鼠脾损伤的作用,可为将来进一步研究提供思路。Objective To detect the expression changes of microRNAs(miRNA)in spleen tissue of septic mice and 24 hours after treated with Qinwen Baidu Decoction by small RNA(sRNA)sequencing technology,and to analyze the possible mechanisms of the anti-septic splenic injury of Qinwen Baidu Decoction at the non-coding gene level.Methods Forty-five SPF healthy BALB/C mice were randomly divided into control group,sepsis group,and sepsis treatment with Qinwen Baidu Decoction group,with 15 mice in each group.Each dose of Qinwen Baidu Decoction contains 15 g of gypsum,7.5 g of rehmannia root,15 g of buffalo horn,2.5 g of coptis chinensis,5 g of gardenia,5 g of platycodon grandiflorum,5 g of scutellaria baicalensis,5 g of anemarrhena,5 g of red peony root,5 g of radix scrophulariae,5 g of forsythia,5 g of licorice,5 g of cortex moutan,and 5 g of bamboo leave.Mice in the sepsis group and sepsis treatment with Qinwen Baidu Decoction group were injected intraperitoneally with lipopolysaccharide(LPS)10 mg/kg to establish sepsis model,and the mice in control group were injected intraperitoneally with equal amounts of normal saline.Mice in the sepsis treatment with Qinwen Baidu Decoction group were received gastric gavage of Qinwen Baidu Decoction 0.3 mL(1 mL was equivalent to 1.5 g of crude drug)2 days before surgery,and twice a day after surgery.Mice in the control group and sepsis group were given equal amounts of normal saline.The spleen tissues of five mice were collected from each group at 24 hours after the operation.The spleen tissues of one mouse in each group were taken to observe the histopathological changes under the light microscope,and the total RNA of the spleen tissues of the remaining mice was extracted.The miRNA was detected by using the sRNA sequencing technology.The difference of miRNA expression and prediction of target genes in the spleen tissues were analyzed using bioinformatics software.Results Light microscope showed that the structure of spleen tissue in the control group was normal.In sepsis group,the overall

关 键 词：脓毒症 脂多糖 小鼠 清瘟败毒饮 微小RNA 

分 类 号：R285.5[医药卫生—中药学]