CAJ | 학술논문

目的探讨肝组织中微小RNA-155(miR-155)表达在脓毒症小鼠肝损伤中的作用.方法按随机数字表法将120只BALB/c小鼠分为脓毒症组和正常对照组,每组60只.腹腔注射脂多糖(LPS)20 mg/kg复制脓毒症模型,术后0、2、6、12、24、48 h每组各取10只小鼠的血及肝组织标本.用酶联免疫吸附试验(ELISA)检测血清和肝组织匀浆中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-6、IL-10)及血清丙氨酸转氨酶(ALT)含量,并观察肝组织病理改变；用实时荧光定量逆转录-聚合酶链反应(RT-PCR)检测肝组织中miR-155的表达.结果脓毒症组血清和肝组织匀浆中TNF-α、IL-6、IL-10水平均随制模时间延长呈升高趋势,TNF-α、IL-6达高峰后逐渐降低,但仍高于对照组水平；TNF-α(ng/L)于2h达高峰(血清:1538.46±102.12比64.52±18.44,肝:255.26±41.23比60.21±13.55),IL-6(μg/L)于6h达高峰(血清:875.33±102.37比153.72±20.67,肝:9.22±0.82比3.35±0.36),IL-10(ng/L)于48 h达高峰(血清:520.13±88.52比23.43±3.01,肝:260.12±50.38比16.37±3.71),与正常对照组比较差异均有统计学意义(均P＜0.05).脓毒症组血清ALT活性(U/L)随制模时间延长逐渐增加,至48 h达高峰(603.26±70.21比45.84±5.64),与正常对照组比较差异有统计学意义(P＜0.05).注射LPS后12h,肝组织结构紊乱,大量炎性细胞浸润,肝细胞水肿、坏死；肝组织中miR-155相对表达量在注射LPS后2h开始升高,12h达高峰,为正常对照组的(72.96±9.34)倍(P＜0.05).结论 MiR-155表达增加在脓毒症肝损伤机制中起重要作用.
목적 탐토간조직중미소RNA-155(miR-155)표체재농독증소서간손상중적작용.방법 안수궤수자표법장120지BALB/c소서분위농독증조화정상대조조,매조60지.복강주사지다당(LPS)20 mg/kg복제농독증모형,술후0、2、6、12、24、48 h매조각취10지소서적혈급간조직표본.용매련면역흡부시험(ELISA)검측혈청화간조직균장중종류배사인자-α(TNF-α)、백세포개소(IL-6、IL-10)급혈청병안산전안매(ALT)함량,병관찰간조직병리개변；용실시형광정량역전록-취합매련반응(RT-PCR)검측간조직중miR-155적표체.결과 농독증조혈청화간조직균장중TNF-α、IL-6、IL-10수평균수제모시간연장정승고추세,TNF-α、IL-6체고봉후축점강저,단잉고우대조조수평；TNF-α(ng/L)우2h체고봉(혈청:1538.46±102.12비64.52±18.44,간:255.26±41.23비60.21±13.55),IL-6(μg/L)우6h체고봉(혈청:875.33±102.37비153.72±20.67,간:9.22±0.82비3.35±0.36),IL-10(ng/L)우48 h체고봉(혈청:520.13±88.52비23.43±3.01,간:260.12±50.38비16.37±3.71),여정상대조조비교차이균유통계학의의(균P＜0.05).농독증조혈청ALT활성(U/L)수제모시간연장축점증가,지48 h체고봉(603.26±70.21비45.84±5.64),여정상대조조비교차이유통계학의의(P＜0.05).주사LPS후12h,간조직결구문란,대량염성세포침윤,간세포수종、배사；간조직중miR-155상대표체량재주사LPS후2h개시승고,12h체고봉,위정상대조조적(72.96±9.34)배(P＜0.05).결론 MiR-155표체증가재농독증간손상궤제중기중요작용.
Objective To evaluate the effects of microRNA-155(miR-155)on liver injury in mice with sepsis.Methods One hundred and twenty BALB/c mice were randomly divided into two groups of equal number according to random number table.Sepsis was induced by intraperitoneal injection of lipopolysaccharide(LPS,20 mg/kg).The mice were sacrificed at the time-points of 0,2,6,12,24,48 hours.Blood and liver tissue were collected,and the levels of tumor necrosis factor-α(TNF-α),interleukin(IL-6,IL-10)in serum and liver homogenate and alanine transaminase(ALT)in serum were determined by enzyme linked immunosorbent assay (ELISA).The injury of liver tissue was evaluated by histopathology.The expression of miR-155 in liver tissue was assessed by fluorescent quantitation reverse transcription-polymerase chain reaction(RT-PCR).Results The levels of TNF-α,IL-6 and IL-10 in serum and liver homogenate of septic mice increased with passage of time,and then the levels of TNF-α and IL-6 lowered after reaching the peak value,but remained higher than that of control group.TNF-α(ng/L)reached the peak value at 2 hours post-LPS-injection(serum:1538.46 ± 102.12 vs.64.52 ± 18.44,liver homogenate:255.26 ± 41.23 vs.60.21 ± 13.55,both P＜0.05).The level of IL-6(μg/L)reached the peak value at 6 hours post-LPS-injection(serum:875.33 ± 102.37 vs.153.72 ± 20.67,liver homogenate:9.22 ± 0.82 vs.3.35 ±0.36,both P＜0.05),and that of IL-10(ng/L)reached the peak value at 48 hours post-LPS-injection(serum:520.13 ± 88.52 vs.23.43 ±3.01,liver homogenate:260.12 ±50.38 vs.16.37 ±3.71,both P＜0.05).There were significant differences in above indexes between septic and control group(all P＜0.05).The serum level of ALT(U/L)rose with passage of time,reaching the peak value at 48 hours post-LPS-injection(603.26 ± 70.21 vs.45.84 ± 5.64,P＜0.05).The values showed significant differences between septic and control group(P＜0.05).A large number of leucocytic infiltration was found in liver.Hepatic tissue showed architectural distortion.Hepatocyte vacuolation and nodular necrosis were obvious at 12 hours post-LPS-injection.Relative expression of miR-155 was found to be increased at 2 hours post-LPS-injection,reaching its peak value at 12 hours post-LPS-injection[(72.96 ± 9.34)-fold of control group,P＜0.05].Conclusion The increase in miR-155 expression might play an important role in the mechanism of liver injury during sepsis.