重组人粒细胞集落刺激因子对缺氧缺血性脑损伤新生大鼠脑组织Nogo受体表达的影响
중조인립세포집락자격인자대결양결혈성뇌손상신생대서뇌조직Nogo수체표체적영향
Effect of recombinant human granulocyte colony-stimulating factor on Nogo receptor expression in the brain tissues of neonatal rats after hypoxic-ischemic brain damage
  • 万方数据
    中华实用儿科临床杂志 중화실용인과림상잡지
    Journal of Applied Clinical Pediatrics
    2015年 6期 456-460 ,共5页

    陈蓉%张晓静%何志旭

    진용%장효정%하지욱

    重组人粒细胞集落刺激因子%缺氧缺血性脑损伤%Nogo受体 重組人粒細胞集落刺激因子%缺氧缺血性腦損傷%Nogo受體
    중조인립세포집락자격인자%결양결혈성뇌손상%Nogo수체
    Recombinant human granulocyte colony-stimulating factor%Hypoxic ischemic brain damage%Nogo receptor
    目的 观察不同时间点2种剂量重组人粒细胞集落刺激因子(rhG-CSF)干预对缺氧缺血性脑损伤(HIBD)新生大鼠脑组织Nogo受体(NgR)表达的影响,揭示rhG-CSF的神经保护作用.方法 将新生7日龄SD大鼠通过抽签法随机分成4组:假手术组、模型组、小剂量组、大剂量组,每组24只.每组分别于手术后1d、3d、7d、14 d处死大鼠,每个时间点各6只.小剂量组、大剂量组分别于造模后即刻颈部皮下注射rhG-CSF50 μg/kg、100 μg/kg,每日1次,连续7d;模型组造模后颈部皮下注射等量9g/L盐水;假手术组不予任何药物.各组新生大鼠于不同时间点取脑组织,采用免疫组织化学法及实时荧光定量PCR法检测脑组织NgR蛋白及NgR mRNA的表达.结果 免疫组织化学:假手术组各时间点大脑皮质均可见NgR蛋白呈基础性表达;与假手术组相比,模型组各时间点NgR蛋白的表达均明显增加(135.67 ±16.63、173.98±17.82、234.00±14.70、319.59±25.22),差异均有统计学意义(P均＜0.01);与模型组相比,小剂量组(134.35±8.89、109.04±12.62、75.99±13.39)和大剂量组(81.38 ±12.25、80.14±10.50、72.58 ±13.66)3 d、7d、14 d大脑皮质NgR蛋白表达均明显下降,差异均有统计学意义(P均＜0.01).大剂量组比小剂量组起效快,大剂量组3d及7d的NgR蛋白表达较小剂量组明显下降,差异均有统计学意义(P均＜0.05).实时荧光定量PCR:与假手术组相比,模型组各时间点NgR mRNA表达量呈增加趋势(1.34±0.24、1.88 ±0.27、2.88 ±0.84、4.26 ±0.86),差异均有统计学意义(P均＜0.05),小剂量组7 d(1.08±0.30)、14 d(0.93 ±0.26)及大剂量组3 d(0.61±0.10)、7 d(0.56 ±0.28)、14 d(0.47±0.12)与模型组相比差异均有统计学意义(P均＜0.05);小剂量组、大剂量组随时间的增加NgR mRNA表达量逐渐下降.大剂量组3d的NgR mRNA表达较小剂量组下降明显(P＜0.05).结论 rhG-CSF干预可降低新生大鼠HIBD模型脑组织NgR的表达,小剂量rhG-CSF干预也可发挥神经保护作用,但其作用可能较弱.
    목적 관찰불동시간점2충제량중조인립세포집락자격인자(rhG-CSF)간예대결양결혈성뇌손상(HIBD)신생대서뇌조직Nogo수체(NgR)표체적영향,게시rhG-CSF적신경보호작용.방법 장신생7일령SD대서통과추첨법수궤분성4조:가수술조、모형조、소제량조、대제량조,매조24지.매조분별우수술후1d、3d、7d、14 d처사대서,매개시간점각6지.소제량조、대제량조분별우조모후즉각경부피하주사rhG-CSF50 μg/kg、100 μg/kg,매일1차,련속7d;모형조조모후경부피하주사등량9g/L염수;가수술조불여임하약물.각조신생대서우불동시간점취뇌조직,채용면역조직화학법급실시형광정량PCR법검측뇌조직NgR단백급NgR mRNA적표체.결과 면역조직화학:가수술조각시간점대뇌피질균가견NgR단백정기출성표체;여가수술조상비,모형조각시간점NgR단백적표체균명현증가(135.67 ±16.63、173.98±17.82、234.00±14.70、319.59±25.22),차이균유통계학의의(P균＜0.01);여모형조상비,소제량조(134.35±8.89、109.04±12.62、75.99±13.39)화대제량조(81.38 ±12.25、80.14±10.50、72.58 ±13.66)3 d、7d、14 d대뇌피질NgR단백표체균명현하강,차이균유통계학의의(P균＜0.01).대제량조비소제량조기효쾌,대제량조3d급7d적NgR단백표체교소제량조명현하강,차이균유통계학의의(P균＜0.05).실시형광정량PCR:여가수술조상비,모형조각시간점NgR mRNA표체량정증가추세(1.34±0.24、1.88 ±0.27、2.88 ±0.84、4.26 ±0.86),차이균유통계학의의(P균＜0.05),소제량조7 d(1.08±0.30)、14 d(0.93 ±0.26)급대제량조3 d(0.61±0.10)、7 d(0.56 ±0.28)、14 d(0.47±0.12)여모형조상비차이균유통계학의의(P균＜0.05);소제량조、대제량조수시간적증가NgR mRNA표체량축점하강.대제량조3d적NgR mRNA표체교소제량조하강명현(P＜0.05).결론 rhG-CSF간예가강저신생대서HIBD모형뇌조직NgR적표체,소제량rhG-CSF간예야가발휘신경보호작용,단기작용가능교약.
    Objective To observe-the different effects of 2 doses recombinant human granulocyte colony-stimulating factors (rhG-CSF) on Nogo receptor(NgR) expression in the brain tissue of neonatal rats after hypoxic-ischemic brain damage(HIBD) at different times in order to reveal the neuroprotective effects of rhG-CSF.Methods Seven-day neonatal Sprague-Dawley(SD) rats were randomly divided into 4 groups by drawing method:sham operation group,model group,low-dose rhG-CSF group and high-dose rhG-CSF group,24 rats in each group.Then each group was divided into 4 subgroups (6 rats in each subgroup)and all rats were exterminated at different times after HIBD(1 d,3 d,7 d and 14 d).In the low-dose rhG-CSF group and high-dose rhG-CSF group,the rats were given daily doses of rhG-CSF 50 μg/kg,100 μg/kg respectively for 7 days by subcutaneous injection immediately after the molding(total 7 injections).In model group,rats received an injection of same amount of 9 g/L saline.In sham operation group,rats received no special treatment.Brain tissues of rats from each group were collected at different time points.The expressions of NgR protein and NgR mRNA in the left brain tissue were detected by immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction (PCR).Results Immunohistochemistry:NgR proteins were constitutively expressed in the cerebral cortex in sham operation group at each time point;compared with sham operation group,the expressions of NgR in model group were increased markedly at each time point (135.67 ± 16.63,173.98 ± 17.82,234.00 ± 14.70,319.59 ± 25.22),and the differences were statistically significant(all P ＜ 0.01);compared with model group,the expressions of NgR in the cerebral cortex in low-dose rhG-CSF group (134.35 ± 8.89,109.04 ± 12.62,75.99 ± 13.39) and high-dose rhG-CSF group (81.38 ± 12.25,80.14 ± 10.50,72.58 ± 13.66) on the 3rd,7th,14th day were reduced significantly (all P ＜ 0.01).Compared with low-dose rhG-CSF group,the protein expressions of NgR in the high-does rhG-CSF group were decreased faster,and had the marked difference on the 3rd,7th day (P ＜ 0.05).Real-time fluorescent quantitative PCR:compared with the sham operation group,the expressions of NgR mRNA increased gradually in the cerebral cortex in the model group (1.34 ± 0.24,1.88 ± 0.27,2.88 ± 0.84,4.26 ± 0.86),the differences in NgR mRNA expression were statistically significant at different times(all P ＜ 0.05) ; compared with model group,the expressions of NgR mRNA in low-dose rhG-CSF group on the 7th (1.08 ± 0.30),14th day (0.93 ± O.26) and high-dose rhG-CSF group on the 3rd (0.61 ± 0.10),7th (0.56 ± 0.28),14th day (0.47 ± 0.12) were significantly different (all P ＜ 0.05).The expressions of low-dose group and high-dose group were reduced gradually.The NgR mRNA expression reduced more quickly in the high-dose group than in the low-dose rhG-CSF group and had substantial difference between two groups in 3 days (P ＜ 0.05).Conclusions The findings suggest that rhG-CSF intervention can reduce the expressions of NgR in the brain tissues of neonatal rats after HIBD,and low-dose rhG-CSF also has neuroprotective effect,but it could be weaker than high-dose rhG-CSF.
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