CAJ | 학술논문

目的探讨雷帕霉素抑制Janus激酶/信号转导和转录激活子(JAK/STAT)通路对急性肝功能衰竭大鼠Toll样受体(TLR)-4基因表达的影响.方法采用腹腔注射D-氨基半乳糖(D-GalN)800 mg/kg和脂多糖(LPS)8 μg/只,建立急性肝功能衰竭大鼠模型,分别在注射D-GalN和LPS后2、6、12、24、48 h 5个时间点留取大鼠血及肝脏标本.SD大鼠分为对照组(n=6)、急性肝功能衰竭模型组(n=30)、STAT抑制剂雷帕霉素(RPM)干预组(n=30),在各不同时间点检测ALT、AST.ELISA法检测血清TNF-α、IL-6水平,RT-PCR法检测大鼠肝组织TLR-4 mRNA表达.数据行t检验.结果急性肝功能衰竭组大鼠在造模后2 h TNF-α、IL-6水平均显著升高,6 h达峰值,RPM可明显抑制TNF-α、IL-6水平.急性肝功能衰竭组大鼠6、12、24、48 h肝组织中TLR-4 mRNA分别为0.745±0.135、1.092±0.175、1.115±0.152和0.812±0.130,RPM干预后分别为0.545±0.118、0.798±0.124、0.857±0.109和0.595±0.152,各时间点两组比较,差异均有统计学意义(t值分别为2.726、3.349、3.382和2.567,均P＜0.05).TLR-4 mRNA表达与ALT、AST均呈正相关(r值分别为0.722、0.712,均P＜0.01).结论抑制JAK/STAT通路可明显下调急性肝功能衰竭大鼠肝组织TLR-4表达,JAK/STAT通路可能参与急性肝功能衰竭过程中TLR-4mRNA表达的调控.
목적 탐토뢰파매소억제Janus격매/신호전도화전록격활자(JAK/STAT)통로대급성간공능쇠갈대서Toll양수체(TLR)-4기인표체적영향.방법채용복강주사D-안기반유당(D-GalN)800 mg/kg화지다당(LPS)8 μg/지,건립급성간공능쇠갈대서모형,분별재주사D-GalN화LPS후2、6、12、24、48 h 5개시간점류취대서혈급간장표본.SD대서분위대조조(n=6)、급성간공능쇠갈모형조(n=30)、STAT억제제뢰파매소(RPM)간예조(n=30),재각불동시간점검측ALT、AST.ELISA법검측혈청TNF-α、IL-6수평,RT-PCR법검측대서간조직TLR-4 mRNA표체.수거행t검험.결과급성간공능쇠갈조대서재조모후2 h TNF-α、IL-6수평균현저승고,6 h체봉치,RPM가명현억제TNF-α、IL-6수평.급성간공능쇠갈조대서6、12、24、48 h간조직중TLR-4 mRNA분별위0.745±0.135、1.092±0.175、1.115±0.152화0.812±0.130,RPM간예후분별위0.545±0.118、0.798±0.124、0.857±0.109화0.595±0.152,각시간점량조비교,차이균유통계학의의(t치분별위2.726、3.349、3.382화2.567,균P＜0.05).TLR-4 mRNA표체여ALT、AST균정정상관(r치분별위0.722、0.712,균P＜0.01).결론억제JAK/STAT통로가명현하조급성간공능쇠갈대서간조직TLR-4표체,JAK/STAT통로가능삼여급성간공능쇠갈과정중TLR-4mRNA표체적조공.
Objective To investigate the effect of rapamycin(RPM) on the gene expression of Toll-like receptor (TLR)-4 by inhibiting Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway in rats with acute liver failure (ALF).Methods The ALF model of rats was induced by D-galactosamine (D-GalN) 800 mg/kg and lipopolysaccharide (LPS) 8 μg.The blood and liver tissue samples were collected at 2,6,12,24 and 48 h after D-GalN/LPS injection.SD rats were randomly divided into three groups:normal control group (n=6),ALF group (n=30) and RPM treatment group (n=30).The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) at each time points were tested.The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 were tested by enzyme-linked immunosorbent assay (ELISA),and the mRNA expressions of TLR-4 in liver tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR).The data analysis was performed using t test.Results The levels of TNF-α and IL-6 were increased markedly 2 h after GalN/LPS iniection and peaked at 6 h.Treatment with RPM could significantly inhibit the levels of TNF-α and IL-6.The TLR-4 mRNA expression levels in liver tissues at 6,12,24,48 h in ALF group were 0.745±0.135,1.092±0.175,1.115±0.152 and 0.812±0.13,respectively;those in RPM group were 0.545±0.118,0.798±0.124,0.857±0.109 and 0.595±0.152,respectively.The differences between two groups at each time points were all significant (t=2.726,3.349,3.382 and 2.567,respectively,all P＜0.05).In addition,TLR-4 mRNA expression was positively correlated with ALT and AST levels(r=0.722 and 0.712,respectively,both P＜0.01).Conclusions Inhibition of JAK/STAT pathway can markedly down regulate TLR-4 gene expression in liver tissue of rats with ALF,which indicates that JAK/STAT pathway may be involved in the regulation of TLR-4 mRNA expression during ALF.