中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2015年
7期
751-755
,共5页
微小RNA-214%小鼠%脓毒症%心肌损伤%炎症因子%心肌酶%凋亡
微小RNA-214%小鼠%膿毒癥%心肌損傷%炎癥因子%心肌酶%凋亡
미소RNA-214%소서%농독증%심기손상%염증인자%심기매%조망
MicroRNA%Sepsis%Myocardial injury%Inflammatory factor%Mycardial enzyme%Apoptosis
目的 探讨微小RNA-214 (miRNA-214)对脓毒症小鼠心肌损伤的作用.方法 清洁级雄性昆明小鼠66只,体质量25 ～30 g,随机(随机数字法)分为4组:假手术组(Sham组,n=18)、脓毒症组(CLP组,n=18)、miRNA-214前体+脓毒症组(前体组,n=12)、miRNA-214抑制剂+脓毒症组(抑制剂组,n=12),其余6只小鼠应用载有miRNA-214前体和抑制剂的腺病毒转染用于空载病对照及miRNA-214转染情况检测.采用盲肠结扎穿孔术(CLP)制备小鼠脓毒症模型.Sham组开腹寻找盲肠,不结扎穿孔;前体组、抑制剂组转染后行CLP术.Sham、CLP组小鼠分别于术后6、12、24h时取心肌组织采用实时荧光定量聚合酶链反应法(RT-PCR)测定miRNA-214表达.于术后12、24h时采用酶联免疫法(ELISA)测定各组心肌组织炎症因子和血清心肌酶浓度;术后24h时采用流式细胞技术测定心肌细胞凋亡率,光镜下观察心肌组织病理学结果.采用SPSS 13.0统计学软件进行分析,应用单因素方差分析进行组间差异比较.结果 (1)与Sham组比较,CLP组术后6、12、24 h miRNA-214表达分别升高2.28倍、1.89倍、1.53倍(F值分别为280.198,82.48,30.65;均P＜0.05);(2)与CLP组术后12 h比较,转染miRNA-214前体小鼠miRNA-214表达水平升高1.86倍,转染抑制剂小鼠miRNA-214表达下降84.5％ (F =394.773;P＜0.05);(3)与CLP组比较,前体组小鼠术后12、24h心肌组织上清液肿瘤坏死因子(TNF-α)、白介素-6(IL-6),血清肌钙蛋白(cTnI)及脑钠肽(BNP)水平降低,白介素-10 (IL-10)水平升高;反之,抑制剂组各指标水平升高,IL-10水平降低(12hF值分别为72.819,88.851,147.546,65.256,102.72;24hF值分别为50.836,76.167,100.290,60.763,52.580;均P＜0.05);(4)与CLP组比较,前体组小鼠术后24h心肌细胞凋亡率下降,抑制剂组升高(F=151.076;P＜0.05);(5)前体组小鼠心肌组织病理学损伤较CLP组、抑制剂组减轻.结论 在CLP所致的小鼠心肌损伤中miRNA-214表达增加,且miRNA-214对CLP诱导的心肌损伤有保护作用.
目的 探討微小RNA-214 (miRNA-214)對膿毒癥小鼠心肌損傷的作用.方法 清潔級雄性昆明小鼠66隻,體質量25 ～30 g,隨機(隨機數字法)分為4組:假手術組(Sham組,n=18)、膿毒癥組(CLP組,n=18)、miRNA-214前體+膿毒癥組(前體組,n=12)、miRNA-214抑製劑+膿毒癥組(抑製劑組,n=12),其餘6隻小鼠應用載有miRNA-214前體和抑製劑的腺病毒轉染用于空載病對照及miRNA-214轉染情況檢測.採用盲腸結扎穿孔術(CLP)製備小鼠膿毒癥模型.Sham組開腹尋找盲腸,不結扎穿孔;前體組、抑製劑組轉染後行CLP術.Sham、CLP組小鼠分彆于術後6、12、24h時取心肌組織採用實時熒光定量聚閤酶鏈反應法(RT-PCR)測定miRNA-214錶達.于術後12、24h時採用酶聯免疫法(ELISA)測定各組心肌組織炎癥因子和血清心肌酶濃度;術後24h時採用流式細胞技術測定心肌細胞凋亡率,光鏡下觀察心肌組織病理學結果.採用SPSS 13.0統計學軟件進行分析,應用單因素方差分析進行組間差異比較.結果 (1)與Sham組比較,CLP組術後6、12、24 h miRNA-214錶達分彆升高2.28倍、1.89倍、1.53倍(F值分彆為280.198,82.48,30.65;均P＜0.05);(2)與CLP組術後12 h比較,轉染miRNA-214前體小鼠miRNA-214錶達水平升高1.86倍,轉染抑製劑小鼠miRNA-214錶達下降84.5％ (F =394.773;P＜0.05);(3)與CLP組比較,前體組小鼠術後12、24h心肌組織上清液腫瘤壞死因子(TNF-α)、白介素-6(IL-6),血清肌鈣蛋白(cTnI)及腦鈉肽(BNP)水平降低,白介素-10 (IL-10)水平升高;反之,抑製劑組各指標水平升高,IL-10水平降低(12hF值分彆為72.819,88.851,147.546,65.256,102.72;24hF值分彆為50.836,76.167,100.290,60.763,52.580;均P＜0.05);(4)與CLP組比較,前體組小鼠術後24h心肌細胞凋亡率下降,抑製劑組升高(F=151.076;P＜0.05);(5)前體組小鼠心肌組織病理學損傷較CLP組、抑製劑組減輕.結論 在CLP所緻的小鼠心肌損傷中miRNA-214錶達增加,且miRNA-214對CLP誘導的心肌損傷有保護作用.
목적 탐토미소RNA-214 (miRNA-214)대농독증소서심기손상적작용.방법 청길급웅성곤명소서66지,체질량25 ～30 g,수궤(수궤수자법)분위4조:가수술조(Sham조,n=18)、농독증조(CLP조,n=18)、miRNA-214전체+농독증조(전체조,n=12)、miRNA-214억제제+농독증조(억제제조,n=12),기여6지소서응용재유miRNA-214전체화억제제적선병독전염용우공재병대조급miRNA-214전염정황검측.채용맹장결찰천공술(CLP)제비소서농독증모형.Sham조개복심조맹장,불결찰천공;전체조、억제제조전염후행CLP술.Sham、CLP조소서분별우술후6、12、24h시취심기조직채용실시형광정량취합매련반응법(RT-PCR)측정miRNA-214표체.우술후12、24h시채용매련면역법(ELISA)측정각조심기조직염증인자화혈청심기매농도;술후24h시채용류식세포기술측정심기세포조망솔,광경하관찰심기조직병이학결과.채용SPSS 13.0통계학연건진행분석,응용단인소방차분석진행조간차이비교.결과 (1)여Sham조비교,CLP조술후6、12、24 h miRNA-214표체분별승고2.28배、1.89배、1.53배(F치분별위280.198,82.48,30.65;균P＜0.05);(2)여CLP조술후12 h비교,전염miRNA-214전체소서miRNA-214표체수평승고1.86배,전염억제제소서miRNA-214표체하강84.5％ (F =394.773;P＜0.05);(3)여CLP조비교,전체조소서술후12、24h심기조직상청액종류배사인자(TNF-α)、백개소-6(IL-6),혈청기개단백(cTnI)급뇌납태(BNP)수평강저,백개소-10 (IL-10)수평승고;반지,억제제조각지표수평승고,IL-10수평강저(12hF치분별위72.819,88.851,147.546,65.256,102.72;24hF치분별위50.836,76.167,100.290,60.763,52.580;균P＜0.05);(4)여CLP조비교,전체조소서술후24h심기세포조망솔하강,억제제조승고(F=151.076;P＜0.05);(5)전체조소서심기조직병이학손상교CLP조、억제제조감경.결론 재CLP소치적소서심기손상중miRNA-214표체증가,차miRNA-214대CLP유도적심기손상유보호작용.
Objective To evaluate the role of miRNA-214 in myocardial injury in septic mice.Methods Sixty-six healthy male Kunming mice,weighing 25-30 g,were randomLy (random number) divided into4 groups:sham operation group (Sham group,n =18),cecal ligation and puncture group (CLP group,n =18),CLP + miRNA-214 precursor treated group (pre-miR-214 group,n =12),CLP + miRNA-214 inhibitor treated (anti-miR-214 group,n =12),and the rest six mice were treated with miRNA-214 precursor or inhibitor intravenously.Sepsis was induced by CLP,pre-miR-214 group and anti-miR-214 group were respectively treated with miRNA-214 precursor or miRNA-214 inhibitor before the CLP,Sham group were exposed to the cecum only.Mice were sacrificed and hearts were removed for determination of miRNA-214 expression by RT-PCR in Sham and CLP group at 6 h,12 h,and 24 h.Blood samples were collected from inferior vena cava at 12 h and 24 h after CLP for determination of the B-type natriuretic peptide (BNP) and cardiac troponin-I (cTnI) concentration by ELISA,then the mice were sacrificed and hearts were removed for determination of the inflammatory factor concentration by ELISA,cardiac myocyte apoptosis rate by flow cytometry and histopathological changes by microscopic examination.Statistical analysis was carried out with SPSS 13.0 software for One-way ANOVA.Results (1) The miRNA-214 expression was higher in CLP group than that in Sham group at 6 h,12 h and 24 h;(2) Compared with CLP group,the miRNA-214 expression in pre-miR-214 group was increased,but decreased in anti-miR-214 group at 12 h;(3) Compared with CLP group,the concentrations of the serum BNP,cTNI,tumor necrosis factor-alpha (TNF-α) and the interleukin-6 (IL-6) were lower than those in pre-miR-214 group,whereas the interleukin-10 (IL-10) was increased.However,the levels of these variables changed just in opposite direction in anti-miR-214 group at 12 h,24 h;(4) Compared with CLP group,the myocardial cell apoptosis rate was decreased in pre-miR-214 group,but increased in anti-miR-214 group at 24 h;(5) The microscopic examination showed that myocardial injury was attenuated in pre-miR-214 group compared with CLP group.Conclusions The miRNA-214 expression was increased in myocardial injury in CLP mice,suggesting miRNA-214 expression relating to myocardial protection in sepsis.
