中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2015年
8期
839-844
,共6页
李芮%崔云%张育才%任玉倩%李江%吕鑫%祝益民
李芮%崔雲%張育纔%任玉倩%李江%呂鑫%祝益民
리예%최운%장육재%임옥천%리강%려흠%축익민
miR-155抑制剂%内毒素血症%肺损伤%JAK/STAT1信号通路%小鼠
miR-155抑製劑%內毒素血癥%肺損傷%JAK/STAT1信號通路%小鼠
miR-155억제제%내독소혈증%폐손상%JAK/STAT1신호통로%소서
MicroRNA-155 inhibitor%Endotoxemia%Lung injury%JAK/STAT1 signaling pathways%Mice
目的 探讨microRNA-155 (miR-155)抑制剂对内毒素血症小鼠肺脏JAK/STAT1信号通路的影响,以及miR-155在内毒素血症诱导肺损伤中的作用.方法 120只BALB/c小鼠按随机数字表法分为内毒素血症组(LPS组)、miR-155抑制组(inhibitor+LPS组)和对照组,每组40只.内毒素血症组腹腔注射LPS 20 mg/kg;miR-155抑制组尾静脉注射miR-155抑制物80 mg/kg 24h后,腹腔注射LPS 20 mg/kg;对照组注射等容量生理盐水.注射LPS6、12、24、48 h后,每组各处死10只小鼠,留取肺脏标本.实时荧光定量聚合酶链反应(qRT-PCR)检测肺组织中miR-155、SOCSI mRNA、STAT1 mRNA表达;酶联免疫吸附试验(EHSA)检测白细胞介素-10(IL-10)和肿瘤坏死因子-α(TNF-α)含量;苏木素-伊红(HE)染色观察肺组织病理学改变.结果 小鼠腹腔注射LPS后,肺组织miR-155表达在24 h内呈升高趋势,48 h时已下降.LPS组各时间点miR-155表达分别为6h (8.52±1.12)、12 h(11.04±0.99)、24h (15.84±0.80)、48 h(4.03 ±2.55).inhibitor+ LPS组miR-155表达量较LPS组低,12 h(t=6.08,P＜0.01)、24h (t=23.64,P＜0.01)差异具有统计学意义.STAT1 mRNA和SOCS1 mRNA表达均在6h最高,后逐渐下降.LPS组STAT1 mRNA高于inhibitor+ LPS组,6h(t=4.41,P＜0.01)、12h(t=2.69,P＜0.05)、24h(=3.62,P＜0.01)差异具有统计学意义.inhibitor+ LPS组SOCS1 mRNA表达高于LPS组,6h(t=4.55,P＜0.01)、12 h(t=4.12,P＜0.01)、24h(t=2.38,P＜0.05)差异具有统计学意义.TNF-α含量于6h最高,IL-10含量于48 h最高.LPS组含量高于对照组和inhibitor+LPS组,6、12、24、48 h差异均有统计学意义(P＜0.01).光镜下观察到inhibitor+ LPS组肺组织病变轻于LPS组.结论 miR-155在内毒素血症小鼠肺组织中表达升高,抑制miR-155能下调JAK/STAT1信号通路,减轻内毒素血症小鼠肺损伤.
目的 探討microRNA-155 (miR-155)抑製劑對內毒素血癥小鼠肺髒JAK/STAT1信號通路的影響,以及miR-155在內毒素血癥誘導肺損傷中的作用.方法 120隻BALB/c小鼠按隨機數字錶法分為內毒素血癥組(LPS組)、miR-155抑製組(inhibitor+LPS組)和對照組,每組40隻.內毒素血癥組腹腔註射LPS 20 mg/kg;miR-155抑製組尾靜脈註射miR-155抑製物80 mg/kg 24h後,腹腔註射LPS 20 mg/kg;對照組註射等容量生理鹽水.註射LPS6、12、24、48 h後,每組各處死10隻小鼠,留取肺髒標本.實時熒光定量聚閤酶鏈反應(qRT-PCR)檢測肺組織中miR-155、SOCSI mRNA、STAT1 mRNA錶達;酶聯免疫吸附試驗(EHSA)檢測白細胞介素-10(IL-10)和腫瘤壞死因子-α(TNF-α)含量;囌木素-伊紅(HE)染色觀察肺組織病理學改變.結果 小鼠腹腔註射LPS後,肺組織miR-155錶達在24 h內呈升高趨勢,48 h時已下降.LPS組各時間點miR-155錶達分彆為6h (8.52±1.12)、12 h(11.04±0.99)、24h (15.84±0.80)、48 h(4.03 ±2.55).inhibitor+ LPS組miR-155錶達量較LPS組低,12 h(t=6.08,P＜0.01)、24h (t=23.64,P＜0.01)差異具有統計學意義.STAT1 mRNA和SOCS1 mRNA錶達均在6h最高,後逐漸下降.LPS組STAT1 mRNA高于inhibitor+ LPS組,6h(t=4.41,P＜0.01)、12h(t=2.69,P＜0.05)、24h(=3.62,P＜0.01)差異具有統計學意義.inhibitor+ LPS組SOCS1 mRNA錶達高于LPS組,6h(t=4.55,P＜0.01)、12 h(t=4.12,P＜0.01)、24h(t=2.38,P＜0.05)差異具有統計學意義.TNF-α含量于6h最高,IL-10含量于48 h最高.LPS組含量高于對照組和inhibitor+LPS組,6、12、24、48 h差異均有統計學意義(P＜0.01).光鏡下觀察到inhibitor+ LPS組肺組織病變輕于LPS組.結論 miR-155在內毒素血癥小鼠肺組織中錶達升高,抑製miR-155能下調JAK/STAT1信號通路,減輕內毒素血癥小鼠肺損傷.
목적 탐토microRNA-155 (miR-155)억제제대내독소혈증소서폐장JAK/STAT1신호통로적영향,이급miR-155재내독소혈증유도폐손상중적작용.방법 120지BALB/c소서안수궤수자표법분위내독소혈증조(LPS조)、miR-155억제조(inhibitor+LPS조)화대조조,매조40지.내독소혈증조복강주사LPS 20 mg/kg;miR-155억제조미정맥주사miR-155억제물80 mg/kg 24h후,복강주사LPS 20 mg/kg;대조조주사등용량생리염수.주사LPS6、12、24、48 h후,매조각처사10지소서,류취폐장표본.실시형광정량취합매련반응(qRT-PCR)검측폐조직중miR-155、SOCSI mRNA、STAT1 mRNA표체;매련면역흡부시험(EHSA)검측백세포개소-10(IL-10)화종류배사인자-α(TNF-α)함량;소목소-이홍(HE)염색관찰폐조직병이학개변.결과 소서복강주사LPS후,폐조직miR-155표체재24 h내정승고추세,48 h시이하강.LPS조각시간점miR-155표체분별위6h (8.52±1.12)、12 h(11.04±0.99)、24h (15.84±0.80)、48 h(4.03 ±2.55).inhibitor+ LPS조miR-155표체량교LPS조저,12 h(t=6.08,P＜0.01)、24h (t=23.64,P＜0.01)차이구유통계학의의.STAT1 mRNA화SOCS1 mRNA표체균재6h최고,후축점하강.LPS조STAT1 mRNA고우inhibitor+ LPS조,6h(t=4.41,P＜0.01)、12h(t=2.69,P＜0.05)、24h(=3.62,P＜0.01)차이구유통계학의의.inhibitor+ LPS조SOCS1 mRNA표체고우LPS조,6h(t=4.55,P＜0.01)、12 h(t=4.12,P＜0.01)、24h(t=2.38,P＜0.05)차이구유통계학의의.TNF-α함량우6h최고,IL-10함량우48 h최고.LPS조함량고우대조조화inhibitor+LPS조,6、12、24、48 h차이균유통계학의의(P＜0.01).광경하관찰도inhibitor+ LPS조폐조직병변경우LPS조.결론 miR-155재내독소혈증소서폐조직중표체승고,억제miR-155능하조JAK/STAT1신호통로,감경내독소혈증소서폐손상.
Objective To investigate the effects of microRNA-155 (miR-155) inhibitor on JAK/STAT1 (Janus kinase/signal transducer and activator transcription 1) signaling pathways in the injured lung tissue induced by lipopolysaccharide (LPS).Methods One hundred and twenty BALB/c mice were randomly divided into control group (n =40),LPS group (n =40),and inhibitor + LPS group (n =40).LPS group and inhibitor + LPS group were made by injection of LPS 20 mg/kg intra-peritonealy,whereas equivalent volume of normal saline was given instead in the control group.The 80 mg/kg of miR-155 inhibitor was injected into caudal vein 24 h before LPS injection in inhibitor + LPS group.Mice were sacrificed at 6 h,12 h,24 h,and 48h separately after LPS injection,and lung tissue were collected.The levels of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) of lung tissue were measured using the enzyme-linked immunosorbent assay (ELISA).Using histopathological examination,the injury of lung tissue was evaluated.The expressions of miR-155,STAT1 mRNA,SOCS1 mRNA in lung tissue were assayed by fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results The miR-155 expression induced by LPS increased at 6 h,12 h,24 h and decreased at 48 h.The miR-155 expressions in LPS group were (8.52 ± 1.12) at 6 h,(11.04 ±0.99) at 12 h,(15.84 ±0.80) at 24 h and (4.03 ± 2.55) at 48 h.In the inhibitor + LPS group,the expressions of miR-155 were lower compared with LPS group,showing significant differences at 12 h (t =6.08,P ＜ 0.01),and at 24 h (t =23.64,P ＜ 0.01).STAT1 mRNA and SOCS1 mRNA both reached peak levels at 6 h after LPS injection.The levels of STAT1 mRNA in LPS group were higher than those in inhibitor + LPS group,showing significant differencesat6h (t=4.41,P＜0.01),12h(t=2.69,P＜0.05),and24h (t=3.62,P＜0.01).The levels of SOCS1 mRNA in inhibitor + LPS group were higher than those in LPS group,showing significant differences at 6 h (t =4.55,P ＜0.01),12 h (t =4.12,P ＜0.01),24 h (t =2.38,P ＜ 0.05).TNF-α reached its peak value at 6 hours and IL-10 reached its peak value at 48 hours.Both TNF-α and IL-10 were higher in LPS group than those in inhibitor + LPS group showing significant differences at 6 h,12 h,24 h (P ＜0.01).The pathologic examination indicated the lung injury in inhibitor + LPS group was milder than that in LPS group.Conclusion The miR-155 increased in lung tissue of endotoxemic mice.miR-155 inhibitor may suppress JAK/STAT1 signaling pathway and protect the lung tissue.
