摘要
目的建立新生大鼠高胆红素血症模型，探讨静脉注入胆红素对脾髓样分化因子88（myeloiddifferentiationfactor88，MyD88）和白细胞介素1受体相关激酶-4（interleukin-1 receptor associ-ated kinase-4，IRAK-4）蛋白表达的影响。方法选用清洁级7～8d新生SD大鼠，雌雄不限，体重12．0～15．0g，采用随机抽签法随机分为空白对照组（I）、脂多糖（1ipopolysaccharide，LPS）对照组（II）、15mg／kg胆红素对照组（11I）、15mr／kg胆红素＋LPS组（1Va）、30mg／kg胆红素＋LPS组（IVb）和50mr／蝇胆红素＋LPS组（IVC），共6组，每组设置2h、5h、24h三个时间点，每个时间点8只。颈静脉注射不同剂量胆红素（15mr／kg、30mr／kg和50mr／kg）建立新生SD大鼠高胆红素血症模型，1h时腹腔注入LPS1mg／kg。在注入胆红素后2h、5h、24h取脾脏，采用免疫组织化学法检测脾MyD88和IRAK-4蛋白的表达。结果（1）LPS可促进脾细胞MyD88和IRAK-4表达（qMyD882h=49．89，qMyD885h=139．54，qIRAK-42h=7．93，qIRK45h=24．30，qIRAK-424h=6．97，P〈0．01）。（2）低浓度胆红素单独作用可促进脾细胞MyD88表达，抑制IRAK-4表达（qMyD882h=0．76，qMyD885h=5．05，qIRAK42h=6．43，qIRAK45h=22．37，qIRAK-424h=1．50，P〈0．01）。（3）低浓度胆红素对LPS刺激MyD88表达的影响不明显（q2h=1．48，q5h=1．45，q24h=0．10，P〉0．05），中、高浓度胆红素对LPS刺激MyD88表达有抑制作用（qⅣb2h=42．87，qⅣc2h=51．38，qⅣb5h=103．61，qⅣc5h=115．44，qⅣb24h=1．18，qⅣc24h=11．66，P〈0．01）。（4）低、中、高浓度胆红素对LPS刺激IRAK-4表达均有抑制作用（qⅣa2h=9．52，qⅣb2h=14．39，qⅣc2h=25．55，qⅣa5h=38．83，qⅣb5h=54．62，qⅣc5h=60．51，qⅣa24h=2．41，qⅣb24h=1．47，qⅣc24h=7．61，P〈0．01）。（5）胆红素对MyD88、IRAK-4的抑制作用，2h时即可观察到，5h时作用明显，24h时低、中浓度胆红素的抑制�
Objective To explore the effects of bilirubin on myeloid differentiation factor 88 （ MyD88 ） and interleukin-1 receptor associated kinase-4 （IRAK-4）. Methods Seven-day-old Sprague Daw- ley rats （clean grade）, male or female, weighing 12. 0 to 15.0 g, were randomly assigned to 6 groups. There were normal saline group（ I ） ,lipopolysaccharide（LPS） control group （LPS, Ⅱ ）, 15 mg/kg bilirubin con- trol （free-LPS） group （Ⅲ） , 15 mg/kg group （Ⅳa） ,30 mg/kg group （Ⅳb） and 50 mg/kg group （Ⅳc） , and then subsequently divided into 2 h,5 h and 24 h subgroups in each groups. Some of the 200 newborn rats died amid the experiment. Finally a total of 144 were involved in the analysis of results, and 8 rats in each subgroups. Newborn Sprague Dawley rats were administered at various doses of bilirubin （15 mg/kg, 30 mg/kg and 50 mg/kg respectively）intravenously; 1 h after injection, the rats were administered LPS intrap-eritoneally at a dose of 1 mg/kg;MyD88 and IRAK-4 were detected by immunohistochemistry at 2 h,5 h and 24 h after the injection of bilirubin. Results （ 1 ） LPS could stimulate the expression of MyD88 and IRAK-4 in spleen cells （ qMyD88 2h = 49. 89, qMyD88 5 h = 139. 54, qlRAK-42h = 7.93, qIRAK-45 h = 24. 30, qlRAK-424h = 6. 97 ,P 〈 0.01 ）. （2） Low concentration of bilirubin could promote the expression of MyD88 and inhibit the ex- pression of IRAK-4 （ qMyD882 h = 0. 76, qMyo885 h = 5. 05, qmAK-42 h = 6. 43, qIRAK-45 h = 22. 37, qmgK-424 h = 1.50, P 〈 0. 01 ）. （3） LPS stimulation MyD88 affected by low concentration of bilirubin was not obvious （ q2 h = 1.48, q5 h = 1.45, q24 h = 0. 10, P 〉 0. 05 ）. Effects of medium and high concentration of bilirubin on LPS stim- ulation MyD88 were inhibitory （ qⅣb2h =42. 87,qⅣc2h = 51.38,qⅣb5h = 103.61 ,qⅣcSh = 115. 44, qⅣb24h = 1. 18, q rvc 24 h = 11. 66, P 〈 0.01 ）. （ 4 ） Effects of low, medium and high concentration of bilirubin on LPS stimulation IRAK-4 were inhibitory �
出处
《中国小儿急救医学》
CAS
2015年第10期699-703,共5页
Chinese Pediatric Emergency Medicine
基金
四川省科技厅科研基金资助项目（05JY029-054-2
07SG004-005）
