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微小RNA-155在脓毒症小鼠肝脏内的表达变化及作用研究

Expression and effects of microRNA-155 in the livers of septic mice

摘要:

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.

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abstracts:

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.

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