中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2013年
1期
1-6
,共6页
王伟静%安方梅%余东山%赵钢德%项晓刚%赖荣陶%桂红莲%郭斯敏%王晖%谢青
王偉靜%安方梅%餘東山%趙鋼德%項曉剛%賴榮陶%桂紅蓮%郭斯敏%王暉%謝青
왕위정%안방매%여동산%조강덕%항효강%뢰영도%계홍련%곽사민%왕휘%사청
肝功能衰竭,急性%小鼠,近交Balb/C%微小核糖核酸%半乳糖胺%脂多糖类%逆转录聚合酶链反应%微阵列分析
肝功能衰竭,急性%小鼠,近交Balb/C%微小覈糖覈痠%半乳糖胺%脂多糖類%逆轉錄聚閤酶鏈反應%微陣列分析
간공능쇠갈,급성%소서,근교Balb/C%미소핵당핵산%반유당알%지다당류%역전록취합매련반응%미진렬분석
Liver failure,acute%Mice,inbred Balb/C%MicroRNAs%Galactosamine%Lipopolysaccharides%Reverse transcriptase polymerase chain reaction Microarray analysis
目的 探讨D氨基半乳糖联合脂多糖诱导急性肝功能衰竭小鼠血清微小核糖核酸(miRNA)的表达变化以及与肝组织miRNA的相关性.方法 Balb/C清洁级小鼠40只分为两组,模型组32只,对照组8只.模型组应用D-氨基半乳糖联合脂多糖诱导Balb/C小鼠急性肝功能衰竭,对照组腹腔注射0.9％氯化钠溶液1 mL.模型组给药后0、3、5和7h分别处死小鼠各8只,采集血清及肝组织,观察小鼠给药后肝功能生物化学指标及肝组织病理学变化.抽提血清及肝组织miRNA,取0、5和7h肝组织进行锁核酸(LNA)-miRNA微阵列分析,并用实时定量反转录PCR检测miRNA.组间均数比较用单因素方差分析,相关性分析采用Pearson和Spearman相关分析.结果 随着小鼠急性肝功能衰竭的发生,肝组织miRNA的表达发生显著变化,共有21个miRNA明显上调,27个miRNA明显下调,其中miRNA-122和miRNA-1187表达下调,miRNA-146a、miRNA-155表达上调.经PCR验证发现小鼠肝组织miRNA 122和miRNA-1187表达逐渐下调,血清miRNA-122和miRNA-1187的表达则逐渐上调；炎性相关的miRNA 146a和miRNA-155在肝组织和血清中表达均明显增加.小鼠miRNA-122和miRNA-1187在组织与血清中的表达呈负相关(r值分别=-0.477和-0.420,P值分别=0.0089和0.0231),而miRNA-155在组织与血清中表达呈正相关(r=0.678,P=0.0001).小鼠血清miRNA 122(r值分别=0.571和0.554)、miRNA-1187(r值分别=0.471和0.542)的相对表达量与ALT和AST水平均呈正相关(均P＜0.05).血清miRNA 122和miRNA-1187在给药5h上升显著,早于血清转氨酶的变化.结论急性肝功能衰竭小鼠肝组织和血清miRNA-122和miRNA-1187的表达呈负相关,血清中miRNA-122、miRNA-1187的变化早于血清转氨酶的变化,有可能成为早期预测肝损伤的血清标志物.
目的 探討D氨基半乳糖聯閤脂多糖誘導急性肝功能衰竭小鼠血清微小覈糖覈痠(miRNA)的錶達變化以及與肝組織miRNA的相關性.方法 Balb/C清潔級小鼠40隻分為兩組,模型組32隻,對照組8隻.模型組應用D-氨基半乳糖聯閤脂多糖誘導Balb/C小鼠急性肝功能衰竭,對照組腹腔註射0.9％氯化鈉溶液1 mL.模型組給藥後0、3、5和7h分彆處死小鼠各8隻,採集血清及肝組織,觀察小鼠給藥後肝功能生物化學指標及肝組織病理學變化.抽提血清及肝組織miRNA,取0、5和7h肝組織進行鎖覈痠(LNA)-miRNA微陣列分析,併用實時定量反轉錄PCR檢測miRNA.組間均數比較用單因素方差分析,相關性分析採用Pearson和Spearman相關分析.結果 隨著小鼠急性肝功能衰竭的髮生,肝組織miRNA的錶達髮生顯著變化,共有21箇miRNA明顯上調,27箇miRNA明顯下調,其中miRNA-122和miRNA-1187錶達下調,miRNA-146a、miRNA-155錶達上調.經PCR驗證髮現小鼠肝組織miRNA 122和miRNA-1187錶達逐漸下調,血清miRNA-122和miRNA-1187的錶達則逐漸上調；炎性相關的miRNA 146a和miRNA-155在肝組織和血清中錶達均明顯增加.小鼠miRNA-122和miRNA-1187在組織與血清中的錶達呈負相關(r值分彆=-0.477和-0.420,P值分彆=0.0089和0.0231),而miRNA-155在組織與血清中錶達呈正相關(r=0.678,P=0.0001).小鼠血清miRNA 122(r值分彆=0.571和0.554)、miRNA-1187(r值分彆=0.471和0.542)的相對錶達量與ALT和AST水平均呈正相關(均P＜0.05).血清miRNA 122和miRNA-1187在給藥5h上升顯著,早于血清轉氨酶的變化.結論急性肝功能衰竭小鼠肝組織和血清miRNA-122和miRNA-1187的錶達呈負相關,血清中miRNA-122、miRNA-1187的變化早于血清轉氨酶的變化,有可能成為早期預測肝損傷的血清標誌物.
목적 탐토D안기반유당연합지다당유도급성간공능쇠갈소서혈청미소핵당핵산(miRNA)적표체변화이급여간조직miRNA적상관성.방법 Balb/C청길급소서40지분위량조,모형조32지,대조조8지.모형조응용D-안기반유당연합지다당유도Balb/C소서급성간공능쇠갈,대조조복강주사0.9％록화납용액1 mL.모형조급약후0、3、5화7h분별처사소서각8지,채집혈청급간조직,관찰소서급약후간공능생물화학지표급간조직병이학변화.추제혈청급간조직miRNA,취0、5화7h간조직진행쇄핵산(LNA)-miRNA미진렬분석,병용실시정량반전록PCR검측miRNA.조간균수비교용단인소방차분석,상관성분석채용Pearson화Spearman상관분석.결과 수착소서급성간공능쇠갈적발생,간조직miRNA적표체발생현저변화,공유21개miRNA명현상조,27개miRNA명현하조,기중miRNA-122화miRNA-1187표체하조,miRNA-146a、miRNA-155표체상조.경PCR험증발현소서간조직miRNA 122화miRNA-1187표체축점하조,혈청miRNA-122화miRNA-1187적표체칙축점상조；염성상관적miRNA 146a화miRNA-155재간조직화혈청중표체균명현증가.소서miRNA-122화miRNA-1187재조직여혈청중적표체정부상관(r치분별=-0.477화-0.420,P치분별=0.0089화0.0231),이miRNA-155재조직여혈청중표체정정상관(r=0.678,P=0.0001).소서혈청miRNA 122(r치분별=0.571화0.554)、miRNA-1187(r치분별=0.471화0.542)적상대표체량여ALT화AST수평균정정상관(균P＜0.05).혈청miRNA 122화miRNA-1187재급약5h상승현저,조우혈청전안매적변화.결론급성간공능쇠갈소서간조직화혈청miRNA-122화miRNA-1187적표체정부상관,혈청중miRNA-122、miRNA-1187적변화조우혈청전안매적변화,유가능성위조기예측간손상적혈청표지물.
Objective To explore the expressions of circulating microRNAs (miRNAs) in acute liver failure mice induced by D-galactosamine (GalN)/lipopolysaccharides (LPS) and the correlation with miRNAs in the liver.Methods Forty clean grade Balb/C mice,with 32 in the model group and 8 in the control group were enrolled in the study.Liver failure was induced by intraperitoneally injection of D-GalN and LPS in mice of the model group,while mice of the control group were intraperitoneally injected with 1 mL 0.9 ％ sodium chloride solution.Serum and liver samples were collected at 0,3,5,7 hours following administration,and eight mice should be supplied to each sample,and changes of alanine aminotransferase (ALT),aspartate aminotransferase (AST) and histopathology of the liver were observed.miRNA from both the serum and the liver was extracted,miRNA expression profile in the liver at 0,5,7 hours by locked nucleic acid (LNA)-miRNA microarray was analyzed and miRNA by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) was detected.Means of the two groups were compared using one-way ANOVA and correlation analyses were performed using Pearson and Spearman correlation.Results Expression of miRNAs in the liver tissue changed significantly over time with the occurrence of acute liver failure in the mice.Twenty-one miRNAs were up-regulated and 27 were down-regulated,among which miRNA-122 and miRNA-1187 were down-regulated while miRNA-146a and miRNA-155 were up-regulated.It was confirmed by the PCR assay that the expression of miRNA-122 and miRNA-1187 in the liver gradually decreased,while those in the serum were up-regulated over time.However,the expressions of inflammation associated miRNA-155 and miRNA-146a were up-regulated both in the serum and the liver after administration.The expressions of miRNA-122 and miRNA-1187 were negatively correlated between serum and liver (r=-0.477,P=0.0089,r=-0.420,P=0.231),while the expressions of miRNA-155 in serum and liver were positively correlated (r=0.678,P=0.0001).Moreover,the expressions of miRNA-122 (r=0.571,0.554) and miRNA-1187 (r=0.471,0.542) were also positively correlated with serum levels of ALT and AST (all P＜0.05).Liver and serum levels of miRNA-122 and miRNA-1187 changed significantly at 5 hours after administration,which preceded the changes of ALT/AST.Conclusions The expressions of miRNA-122 and miRNA-1187 in serum are well inversely correlated with the corresponding expressions in liver tissues during acute liver failure in mice.The changes of miRNA-122 and miRNA-1187 in the serum precede those of ALT/AST.These data suggest that serum miRNA-122 and miRNA-1187 might be the candidate serum biomarkers for early prediction of liver injury.
