中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2014年
17期
1325-1330
,共6页
徐发林%郭佳佳%王彩红%朱慧英%张彦华%宋娟%段佳佳
徐髮林%郭佳佳%王綵紅%硃慧英%張彥華%宋娟%段佳佳
서발림%곽가가%왕채홍%주혜영%장언화%송연%단가가
促红细胞生成素%促红细胞生成素受体%髓鞘碱性蛋白%胶质纤维酸性蛋白%大鼠,新生
促紅細胞生成素%促紅細胞生成素受體%髓鞘堿性蛋白%膠質纖維痠性蛋白%大鼠,新生
촉홍세포생성소%촉홍세포생성소수체%수초감성단백%효질섬유산성단백%대서,신생
Erythropoietin%Erythropoietin receptor%Myelin basic protein%Glial fibrillary acidic protein%Neonatal rat
目的 探讨促红细胞生成素(EPO)对感染致新生大鼠脑损伤保护作用的最佳应用时机及其相关机制.方法 2日龄(P2)新生SD大鼠按随机数字表法分为4组,分别为对照组(A组)、脂多糖(LPS)感染组(B组)、早期EPO干预组(C组)、晚期EPO干预组(D组).A、B、C组P2新生大鼠连续5d(P2-P6)分别腹腔注射相应药物:等容积9 g/L盐水+等容积EPO空白对照品、0.6 mg/kg LPS+等容积EPO空白对照品、0.6mg/kg LPS+5 000 IU/kg EPO;D组P2新生大鼠连续5d(P2-P6)腹腔注射0.6 mg/kg LPS,P7开始连续腹腔注射5 000 IU/kg EPO 5 d(即P7-P11).A、B2组分别于P2(腹腔注射第1次药物后6h)、P7、P12,各随机数字表法抽取10只新生大鼠取脑,以矢状缝为标志分为左右半脑,右侧脑应用酶联免疫吸附法检测脑组织EPO受体(EPOR)水平,左侧脑应用反转录-聚合酶链反应(RT-PCR)方法检测EPOR mRNA水平;A、B、C3组于P7随机数字表法选取10只新生大鼠灌注取脑,余续养,4组均于P12灌注取脑,应用免疫组织化学方法检测髓鞘碱性蛋白(MBP)、胶质纤维酸性蛋白(GFAP)及EPOR的表达,采用HE染色观察各组大鼠脑组织的病理改变.结果 1.HE染色示B组海马锥体细胞界限不清、层次紊乱,细胞数目减少,脑室扩张,周围白质有囊性软化区域形成;EPO干预组较B组病理改变减轻,早期干预组更明显.2.B组较A组EPOR蛋白及mRNA表达增加,随日龄的增加EPOR与EPOR mRNA表达有下降趋势.3.B组MBP表达(107.46±3.65)较A组(146.78±3.13)明显减少(P＜0.05),EPO干预组较B组表达增加,且C组(126.25±4.42)较D组(117.35±3.42)增加更明显(P＜0.05).4.B组GFAP表达(P7、P12分别为141.46±11.92、149.48±13.59)较A组(P7、P12分别为120.63±13.32、119.74±12.48)增加(P＜0.05),P12时EPO干预组表达较B组降低,C组(134.59±12.19)与D组(137.27±13.87)差异无统计学意义(P＞0.05).结论 EPO对出生后感染所致的脑白质损伤有保护作用,且甲期应用优于晚期,其机制可能与感染可使新生大鼠脑组织EPOR表达增加及EPOR表达随日龄增加而降低有关.
目的 探討促紅細胞生成素(EPO)對感染緻新生大鼠腦損傷保護作用的最佳應用時機及其相關機製.方法 2日齡(P2)新生SD大鼠按隨機數字錶法分為4組,分彆為對照組(A組)、脂多糖(LPS)感染組(B組)、早期EPO榦預組(C組)、晚期EPO榦預組(D組).A、B、C組P2新生大鼠連續5d(P2-P6)分彆腹腔註射相應藥物:等容積9 g/L鹽水+等容積EPO空白對照品、0.6 mg/kg LPS+等容積EPO空白對照品、0.6mg/kg LPS+5 000 IU/kg EPO;D組P2新生大鼠連續5d(P2-P6)腹腔註射0.6 mg/kg LPS,P7開始連續腹腔註射5 000 IU/kg EPO 5 d(即P7-P11).A、B2組分彆于P2(腹腔註射第1次藥物後6h)、P7、P12,各隨機數字錶法抽取10隻新生大鼠取腦,以矢狀縫為標誌分為左右半腦,右側腦應用酶聯免疫吸附法檢測腦組織EPO受體(EPOR)水平,左側腦應用反轉錄-聚閤酶鏈反應(RT-PCR)方法檢測EPOR mRNA水平;A、B、C3組于P7隨機數字錶法選取10隻新生大鼠灌註取腦,餘續養,4組均于P12灌註取腦,應用免疫組織化學方法檢測髓鞘堿性蛋白(MBP)、膠質纖維痠性蛋白(GFAP)及EPOR的錶達,採用HE染色觀察各組大鼠腦組織的病理改變.結果 1.HE染色示B組海馬錐體細胞界限不清、層次紊亂,細胞數目減少,腦室擴張,週圍白質有囊性軟化區域形成;EPO榦預組較B組病理改變減輕,早期榦預組更明顯.2.B組較A組EPOR蛋白及mRNA錶達增加,隨日齡的增加EPOR與EPOR mRNA錶達有下降趨勢.3.B組MBP錶達(107.46±3.65)較A組(146.78±3.13)明顯減少(P＜0.05),EPO榦預組較B組錶達增加,且C組(126.25±4.42)較D組(117.35±3.42)增加更明顯(P＜0.05).4.B組GFAP錶達(P7、P12分彆為141.46±11.92、149.48±13.59)較A組(P7、P12分彆為120.63±13.32、119.74±12.48)增加(P＜0.05),P12時EPO榦預組錶達較B組降低,C組(134.59±12.19)與D組(137.27±13.87)差異無統計學意義(P＞0.05).結論 EPO對齣生後感染所緻的腦白質損傷有保護作用,且甲期應用優于晚期,其機製可能與感染可使新生大鼠腦組織EPOR錶達增加及EPOR錶達隨日齡增加而降低有關.
목적 탐토촉홍세포생성소(EPO)대감염치신생대서뇌손상보호작용적최가응용시궤급기상관궤제.방법 2일령(P2)신생SD대서안수궤수자표법분위4조,분별위대조조(A조)、지다당(LPS)감염조(B조)、조기EPO간예조(C조)、만기EPO간예조(D조).A、B、C조P2신생대서련속5d(P2-P6)분별복강주사상응약물:등용적9 g/L염수+등용적EPO공백대조품、0.6 mg/kg LPS+등용적EPO공백대조품、0.6mg/kg LPS+5 000 IU/kg EPO;D조P2신생대서련속5d(P2-P6)복강주사0.6 mg/kg LPS,P7개시련속복강주사5 000 IU/kg EPO 5 d(즉P7-P11).A、B2조분별우P2(복강주사제1차약물후6h)、P7、P12,각수궤수자표법추취10지신생대서취뇌,이시상봉위표지분위좌우반뇌,우측뇌응용매련면역흡부법검측뇌조직EPO수체(EPOR)수평,좌측뇌응용반전록-취합매련반응(RT-PCR)방법검측EPOR mRNA수평;A、B、C3조우P7수궤수자표법선취10지신생대서관주취뇌,여속양,4조균우P12관주취뇌,응용면역조직화학방법검측수초감성단백(MBP)、효질섬유산성단백(GFAP)급EPOR적표체,채용HE염색관찰각조대서뇌조직적병리개변.결과 1.HE염색시B조해마추체세포계한불청、층차문란,세포수목감소,뇌실확장,주위백질유낭성연화구역형성;EPO간예조교B조병리개변감경,조기간예조경명현.2.B조교A조EPOR단백급mRNA표체증가,수일령적증가EPOR여EPOR mRNA표체유하강추세.3.B조MBP표체(107.46±3.65)교A조(146.78±3.13)명현감소(P＜0.05),EPO간예조교B조표체증가,차C조(126.25±4.42)교D조(117.35±3.42)증가경명현(P＜0.05).4.B조GFAP표체(P7、P12분별위141.46±11.92、149.48±13.59)교A조(P7、P12분별위120.63±13.32、119.74±12.48)증가(P＜0.05),P12시EPO간예조표체교B조강저,C조(134.59±12.19)여D조(137.27±13.87)차이무통계학의의(P＞0.05).결론 EPO대출생후감염소치적뇌백질손상유보호작용,차갑기응용우우만기,기궤제가능여감염가사신생대서뇌조직EPOR표체증가급EPOR표체수일령증가이강저유관.
Objective To explore the influence of erythropoietin (EPO) on infection induced neonatal rat brain injury at different starting time and its related mechanism.Methods Postnatal day 2 (P2) newborn SD rats were randomly divided into 4 groups:control group (group A),lipopolysaccharide (LPS) group (group B),the early EPO group(group C)and the later EPO group(group D).Pups in group A,B and C were injected different drugs intraperitoneally(group A for saline,group B for 0.6 mg/kg of LPS,and group C for 0.6 mg/kg of LPS and 5 000 U/kg of EPO) once a day for consecutive 5 days(P2-P6).LPS in group D were injected 0.6 mg/kg of LPS intraperitoneally once a day for consecutive 5 days(P2 P6),and with 5 000 U/kg of EPO once a day for consecutive 5 days(P7-P1 1).Rats in each group were given different drugs starting at corresponding time by intraperitoneal injection for 5 consecutive days.Every 10 newborn rats in group A and B were selected randomly on P2(6 h after intraperitoneal injection of drugs for the first time),P7 and P12,the brains were divided into the left and the right hemispheres marked by sagittal suture,using enzyme-linked immunosorbent assay method to evaluate the erythropoietin receptor(EPOR) protein level with the right cerebral hemisphere and reverse transcription-polymerase chain reaction (RT-PCR) method was used to investigate EPOR mRNA level of the left cerebral hemisphere.Immunohistochemical method was adopted to evaluate the expression of myelin basic protein(MBP),glial fibrillary acidic protein(GFAP) and EPOR at specified time point,and HE dyeing for the pathological changes of brain damage in different groups.Results HE staining of the group A presented the normal structure in the neonatal rat brain.Reduced numbers of hippocampal pyramidal cells,expansion of the lateral ventricles and periventricular leukomalacia were found in group B.No leukomalacia or lateral ventricles's expansion in EPO administrated groups and it was more obvious in group C.The EPOR protein and mRNA of group B was increased compared with the group A.The EPOR protein and mRNA levels had a tendency to decline with the increase of age.The MBP expression of group B(107.46 ±3.65)was significantly reduced compared with the group A(146.78 ± 3.13) (P ＜ 0.05),and the expression of EPO groups increased in contrast to the group B,moreover,the group C (126.25 ± 4.42) increased more obviously than that of group D(117.35 ± 3.42) (P ＜ 0.05).The GFAP expression of group B(141.46 ± 11.92 at P7 and 149.48 ± 13.59 at P12) increased significantly than group A(120.63 ± 13.32 at P7 and 119.74 ± 12.48 at P12) (P ＜0.05),the EPO group expressed lower than group B at the P12,and the group C (134.59 ± 12.19) decreased than the group D(137.27 ± 13.87) (P ＞ 0.05).Conclusions EPO shows a protective effect on the cerebral white matter injury caused by postpartum infection,it is superior to administer EPO at early time than later time.The mechanism of the protective effect may be connected with the fact that the infection can induce the expression of brain EPOR and the EPOR expression level has a tendency to decline with the increase of age.
