缺氧缺血,脑%大鼠%米诺环素%白质软化病,脑室周围
缺氧缺血,腦%大鼠%米諾環素%白質軟化病,腦室週圍
결양결혈,뇌%대서%미낙배소%백질연화병,뇌실주위
Hypoxia-ischemia,brain%Rats%Minocycline%Leukomalacia,periventricular
目的 研究米诺环素(minocycline,MN)对未成熟新生大鼠缺氧缺血性脑损伤(hypoxic-ischemic brain damage,HIBD)的作用.方法 生后2 d(P2)Sprague-Dawley(SD)大鼠192只,随机分成正常对照组、假手术组、HIBD组、HIBD+MN组,每组48只.通过结扎左侧颈总动脉及8%氮氧混合气缺氧4 h,制作未成熟新生大鼠缺氧缺血性脑损伤模型.假手术组只分离左侧颈总动脉,正常组未予任何处理.HIBD+MN组动物于HI后腹腔注射米诺环素45 mg/kg,此后2 d每24小时腹腔注射1次.HI后3 d,1、2、4周,行常规HE染色及病理评分,HI后3 d及2周,行抗O4、抗O1免疫组化及髓鞘脂碱性蛋白(MBP)免疫组化.HI后4周行神经行为学检测.结果 HIBD组病理可见脑室周围白质损伤,HIBD+MN组病变较轻;O4阳性细胞计数:HIBD+MN组与正常组差异无统计学意义,P＞0.05,而HIBD组较正常组,假手术组、HIBD+MN组减少,差异有统计学意义(23.67±12.00 vs 52.89±10.68,39.28±11.78,41.63±8.41,P＜0.05);O1阳性细胞计数:正常组,假手术组、HIBD组、HIBD+MN组组间差异无统计学意义,P=0.093;MBP阳性纤维束计数:HIBD+MN组与正常组、假手术组差别有统计学意义,P＜0.05,而HIBD组较正常组、假手术组、HIBD+MN组减少,差异有统计学意义(14.71 ±7.42 vs 36.67±6.50,35.50±3.24,26.33±5.92,P＜0.05);HIBD组HI后4周出现明显远期行为学异常,在斜坡试验、悬吊试验及Cylinder试验中,HIBD+MN组与正常组、假手术组差别无统计学意义,P＞0.05;HIBD组与正常组、假手术组及HIBD+MN组差异有统计学意义,P＜0.05;在旷场试验中,HIBD+MN组与HIBD组差异无统计学意义,P=0.772,两组与正常组,假手术组差异均有统计学意义,P＜0.05.结论 米诺环素对未成熟新生大鼠缺氧缺血性脑损伤有早期及远期保护作用.
目的 研究米諾環素(minocycline,MN)對未成熟新生大鼠缺氧缺血性腦損傷(hypoxic-ischemic brain damage,HIBD)的作用.方法 生後2 d(P2)Sprague-Dawley(SD)大鼠192隻,隨機分成正常對照組、假手術組、HIBD組、HIBD+MN組,每組48隻.通過結扎左側頸總動脈及8%氮氧混閤氣缺氧4 h,製作未成熟新生大鼠缺氧缺血性腦損傷模型.假手術組隻分離左側頸總動脈,正常組未予任何處理.HIBD+MN組動物于HI後腹腔註射米諾環素45 mg/kg,此後2 d每24小時腹腔註射1次.HI後3 d,1、2、4週,行常規HE染色及病理評分,HI後3 d及2週,行抗O4、抗O1免疫組化及髓鞘脂堿性蛋白(MBP)免疫組化.HI後4週行神經行為學檢測.結果 HIBD組病理可見腦室週圍白質損傷,HIBD+MN組病變較輕;O4暘性細胞計數:HIBD+MN組與正常組差異無統計學意義,P＞0.05,而HIBD組較正常組,假手術組、HIBD+MN組減少,差異有統計學意義(23.67±12.00 vs 52.89±10.68,39.28±11.78,41.63±8.41,P＜0.05);O1暘性細胞計數:正常組,假手術組、HIBD組、HIBD+MN組組間差異無統計學意義,P=0.093;MBP暘性纖維束計數:HIBD+MN組與正常組、假手術組差彆有統計學意義,P＜0.05,而HIBD組較正常組、假手術組、HIBD+MN組減少,差異有統計學意義(14.71 ±7.42 vs 36.67±6.50,35.50±3.24,26.33±5.92,P＜0.05);HIBD組HI後4週齣現明顯遠期行為學異常,在斜坡試驗、懸弔試驗及Cylinder試驗中,HIBD+MN組與正常組、假手術組差彆無統計學意義,P＞0.05;HIBD組與正常組、假手術組及HIBD+MN組差異有統計學意義,P＜0.05;在曠場試驗中,HIBD+MN組與HIBD組差異無統計學意義,P=0.772,兩組與正常組,假手術組差異均有統計學意義,P＜0.05.結論 米諾環素對未成熟新生大鼠缺氧缺血性腦損傷有早期及遠期保護作用.
목적 연구미낙배소(minocycline,MN)대미성숙신생대서결양결혈성뇌손상(hypoxic-ischemic brain damage,HIBD)적작용.방법 생후2 d(P2)Sprague-Dawley(SD)대서192지,수궤분성정상대조조、가수술조、HIBD조、HIBD+MN조,매조48지.통과결찰좌측경총동맥급8%담양혼합기결양4 h,제작미성숙신생대서결양결혈성뇌손상모형.가수술조지분리좌측경총동맥,정상조미여임하처리.HIBD+MN조동물우HI후복강주사미낙배소45 mg/kg,차후2 d매24소시복강주사1차.HI후3 d,1、2、4주,행상규HE염색급병리평분,HI후3 d급2주,행항O4、항O1면역조화급수초지감성단백(MBP)면역조화.HI후4주행신경행위학검측.결과 HIBD조병리가견뇌실주위백질손상,HIBD+MN조병변교경;O4양성세포계수:HIBD+MN조여정상조차이무통계학의의,P＞0.05,이HIBD조교정상조,가수술조、HIBD+MN조감소,차이유통계학의의(23.67±12.00 vs 52.89±10.68,39.28±11.78,41.63±8.41,P＜0.05);O1양성세포계수:정상조,가수술조、HIBD조、HIBD+MN조조간차이무통계학의의,P=0.093;MBP양성섬유속계수:HIBD+MN조여정상조、가수술조차별유통계학의의,P＜0.05,이HIBD조교정상조、가수술조、HIBD+MN조감소,차이유통계학의의(14.71 ±7.42 vs 36.67±6.50,35.50±3.24,26.33±5.92,P＜0.05);HIBD조HI후4주출현명현원기행위학이상,재사파시험、현조시험급Cylinder시험중,HIBD+MN조여정상조、가수술조차별무통계학의의,P＞0.05;HIBD조여정상조、가수술조급HIBD+MN조차이유통계학의의,P＜0.05;재광장시험중,HIBD+MN조여HIBD조차이무통계학의의,P=0.772,량조여정상조,가수술조차이균유통계학의의,P＜0.05.결론 미낙배소대미성숙신생대서결양결혈성뇌손상유조기급원기보호작용.
Objective To establish a model of immature rat hypoxic-ischemic brain damage (HIBD) which was expected to be similar to periventricular leukomalacia in human preterm infants pathologically and neuroethologically, and to investigate the role of minocycline (MN) in this model.Method Totally 192 Sprague-Dawley rats ( postnatal day 2, P2), of either sex, were randomly divided into 4 groups: normal-group, sham operation group, HIBD-group, HIBD + MN group, each group had 48 rats. HIBD group and HIBD + MN group survived the left common carotid artery (CCA) ligation followed by 4h exposure to 8% O2. Rats in sham operation group only survived the left CCA isolation. Rats in normal group were not treated with anything. In HIBD + MN group, the rats were treated with intraperitoneal injection of minocycline 45 mg/kg, immediately after HI and every 24 h for 2 days. Brain tissues were collected on day 3, 1 week, 2 weeks, 4 weeks after HI, for hematoxylin-eosin staining and histological scoring. Frozen sections of the brains were stained with anti-O4, anti-O1 immunohistochemistry on day 3 after HI, and MBP immunohistochemistry 2 weeks after HI. Rats in the four groups underwent neuroethologic examination 4 weeks after HI. Result In the HIBD group, there were pathological changes in the periventricular white matter. The pathological changes were milder in HIBD + MN group; There was no statistically significan defference between the normal group and HIBD + MN group in the number of positively stained O4 cell (P ＞0.05 ). The number of positively stained O4 cell in the HIBD group was significantly reduced, compared with that of normal group, sham operation group, and HIBD + MN group (23.67 ± 12.00 vs. 52. 89 ± 10.68,39.28 ± 11.78,41.63 ± 8.41, P ＜ 0.05 ). The differences in the number of positively stained O1 cell among the normal group, sham operation group, HIBD group and HIBD + MN group had no statistical significance(P = 0.093). The numbers of myelin basic protein (MBP) positively immunostained fiber bundles in the HIBD + MN group were significantly less than that of the normal group and sham operation group (P＜0.05). The numbers of MBP positively immunostained fiber bundles in the HIBD group were significantly less than that of the normal group, sham operation group, and HIBD + MN group ( 14.71 ±7.42 vs. 36.67 ± 6.50, 35.50 ± 3.24, 26.33 ± 5.92, P ＜ 0.05 ). The HIBD group had long-term neuroethologic abnormality. There was no statistically significant difference in the inclined plane test,hanging test and cylinder test among the HIBD + MN group, normal group, and sham operation group (P ＞0.05). The scores of the HIBD group had statistical significance among the normal group, sham operation group and HIBD + MN group (P ＜ 0.05). In the open field test, there was no statistically significan difference between the HIBD group and HIBD + MN group (P =0.772), but there was significant difference between these two groups and the normal group, sham operation group (P ＜0.05). Conclusion Minocycline protects the pre-oligodendrocyte and has protective effects in terms of long-term neuroethology.
