脑卒中后抑郁大鼠海马原位增殖的新生细胞存活和分化状态

目的 观察脑卒中后抑郁(post-stroke depression,PSD)大鼠海马原位增殖新生细胞的存活和分化.方法 采用左侧大脑中动脉阻塞(MCAO)联合慢性不可预见温和应激刺激(chronicmild stress,CMS)及孤养法建立PSD模型,将雄性SD大鼠分为假手术、脑卒中、CMS和PSD组.每组均为18只.采取免疫组织化学、荧光双标染色及共聚焦成像动态检测,比较各研究组大鼠左侧海马齿状回溴脱氧尿苷嘧啶(BrdU)及其与神经元特异性核蛋白(NeuN)或胶质纤维酸性蛋白(GFAP)共表达.结果 与脑卒中组(232.2±8.6、123.7±2.6、136.2±2.6)相比,PSD组大鼠左侧(伤侧)海马齿状回BrdU+细胞数在脑梗死后第21(156.2±2.5)、30(70.2±2.0)和45天(81.2±1.1)均明显减少(t=28.83、52.2、62.08,均P<0.01),但仍高于假手术组.与脑卒中组(79.3%±2.8%、87.7%±4.6%)相比,PSD组大鼠左侧(伤侧)海马齿状回BrdU+/NeuN+细胞比例在脑梗死后第30(69.0%±3.4%)和45天(78.3%±2.4%)均明显减少(t=5.871、4.403,均P<0.01).BrdU+/GFAP+细胞比例在脑梗死后30和45 d均明显增加(t=4.226、8.945,P<0.01).结论 PSD大鼠脑卒中后海马齿状回原位增殖的新生细胞存活降低,分化为神经元的比例下降,胶质细胞比例增加.     

卡马西平对匹罗卡品诱导成年癫间疒大鼠海马齿状回神经前体细胞增殖的影响

目的研究卡马西平对成年癫大鼠海马齿状回内源性神经前体细胞增殖的影响。方法采用氯化锂和匹罗卡品联合诱导大鼠癫模型,将癫大鼠随机分为癫对照组和癫卡马西平组,正常大鼠随机分为正常对照组和正常卡马西平组。癫对照组和正常对照组给以蒸馏水灌胃,同时癫卡马西平组和正常卡马西平组给予卡马西平灌胃。于灌胃后第6d腹腔注射5-溴脱氧尿苷嘧啶(BrdU)标记海马齿状回的内源性神经前体细胞的增殖情况;用免疫组化方法观察各组大鼠在注射BrdU后第1d、第7d齿状回BrdU阳性细胞数量的表达。结果①注射BrdU后第1d,癫对照组大鼠海马齿状回BrdU阳性细胞数较正常对照组明显增加(P<0.01),癫卡马西平组大鼠海马齿状回BrdU阳性细胞数较癫对照组减少(P<0.05);②注射BrdU后第7d,癫对照组大鼠海马齿状回BrdU阳性细胞数较正常对照组明显增加(P<0.01),癫卡马西平组大鼠海马齿状回BrdU阳性细胞数较癫对照组明显减少(P<0.05)。结论卡马西平抑制癫大鼠海马齿状回内源性神经前体细胞增殖。     

复智散对阿尔茨海默病大鼠海马内源性神经干细胞增殖的影响

目的观察复智散（FZS）对Alzheimer病（AD）模型大鼠海马齿状回内源性神经干细胞增殖的影响。方法采用β淀粉样蛋白25—35（Aβ25—35）侧脑室注射制作AD大鼠模型。采用Morris水迷宫检测大鼠的学习记忆能力,免疫荧光检测大鼠海马5-溴脱氧尿嘧啶（BrdU）阳性细胞表达,免疫组化检测海马增殖细胞核抗原（PCNA）阳性细胞表达,并对海马齿状回下颗粒层、海马门、分子层分别进行BrdU、PCNA阳性细胞计数。结果与模型组比较,健康对照组、假手术组和FZS治疗组大鼠水迷宫实验中的平均逃避潜伏期缩短（P〈0．05）,齿状回颗粒细胞下层BrdU、PCNA阳性细胞数量明显增加（P〈0．05）,但后三组之间比较差异无统计学意义。结论FZS可促进AD模型大鼠海马齿状回内源性神经干细胞增殖。     

不同程度的性发作对成年大鼠空间学习记忆影响的研究

目的研究不同程度的性发作对成年大鼠空间学习记忆的影响。方法采用氯化锂和匹罗卡品联合诱导大鼠不同程度的癫模型(轻型和重型)。于造模后第6d给所有大鼠腹腔注射5-溴脱氧尿苷嘧啶(BrdU+)标记海马齿状回增殖的内源性神经前体细胞;用免疫组化方法观察各组大鼠注射BrdU+后第1d和第28d齿状回BrdU+阳性细胞数以及第28d的BrdU+/神经元核性蛋白(NeuN+)阳性细胞数及分布情况;利用Morris水迷宫评价大鼠的学习记忆功能。结果与正常组及轻型组比较,在各个时间点重型组海马齿状回BrdU+细胞数均增加(P<0.05),28d时BrdU+/NeuN+细胞数相应增多,但其占BrdU+细胞数的比例明显下降(P<0.05)。28d时重型组大鼠的学习记忆功能较正常组及轻型组明显下降(P<0.05)。结论严重的癫发作造成大鼠对空间学习记忆功能的损害,可能与其刺激大鼠海马齿状回内源性神经前体细胞增殖水平,抑制其分化为新生的成熟神经元有关。     

老年大鼠脑出血后海马齿状回神经干细胞的增殖与分化

目的 观察老年大鼠脑出血后海马齿状回神经干细胞(NSCs)的增殖与分化,探讨脑出血后NSCs的变化规律.方法 制作老年大鼠脑出血模型,5-溴脱氧尿核苷(BrdU)腹腔注射标记增殖细胞,用免疫组化法检测大鼠海马齿状回BrdU、神经元核抗原(NeuN)、胶质纤维酸性蛋白(GFAP)阳性细胞数的变化.结果 正常组和假手术组老年大鼠海马齿状回均有少量BrdU阳性细胞,脑出血后大鼠各时间段的BrdU阳性细胞数目均较正常组和假手术组明显增加,7d组达到峰值后逐渐下降,28d组仍高于正常组和假手术组.正常老年大鼠海马齿状回可见少量BrdU/NeuN和BrdU/GFAP双标阳性细胞,脑出血后双标阳性细胞数较正常组明显增加.结论 脑出血后老年大鼠海马齿状回NSCs增殖明显,且可以向神经元和神经胶质细胞分化.     

rTMS促进大鼠局灶性脑缺血海马内源性神经干细胞分化的研究

目的探讨重复经颅磁刺激（rTMS）对局灶性脑缺血大鼠海马内源性神经干细胞分化的影响。方法线栓法制备大鼠大脑中动脉闭塞（MCAO）模型,随机分为脑缺血自然恢复组和rTMS治疗组,用荧光显微镜和共聚焦显微镜观察缺血14d、28d后各组大鼠海马中5-溴脱氧尿核苷（BrdU）与神经元特异核蛋白（NeuN）、神经胶质酸性蛋白（GFAP）共同标记的阳性细胞,并在高倍荧光显微镜下对双标阳性细胞计数。结果脑缺血后14d、28d,rTMS治疗组大鼠海马BrdU／NeuN双标阳性细胞数量分别为17．12±2．91、23．20±5．97,较相应自然恢复组12．96±2．79、15．92±2．52明显增加,两组同一时间点组间比较有统计学差异（P〈0．01）。而脑缺血后14d、28d,rTMS治疗组大鼠海马BrdU／GFAP双标阳性细胞数量分别为30．48±4．58、36．48±4．90,较相应自然恢复组37．44±3．58、43．60±5．96减少,两者相比有统计学差异（P〈0．05）。结论局灶性脑缺血大鼠海马增殖的内源性神经干细胞,可分化为神经元或神经胶质细胞,而rTMS可促进海马内源性神经干细胞向神经元的分化。     

Orexin-1受体拮抗剂对慢性点燃癫大鼠学习记忆的影响及海马神经细胞的增殖变化

目的:研究orexin-1受体(OX1R)拮抗剂(SB334867,SB)对戊四氮(PTZ)慢性点燃癫大鼠空间学习记忆能力及海马齿状回神经细胞增殖的影响。方法:Wistar大鼠随机分为①对照组[腹腔和侧脑室均注射生理盐水(NS)];②PTZ组(腹腔注射PTZ+侧脑室注射NS);③PTZ+orexin-A(OXA)组(腹腔注射PTZ+侧脑室注射OXA);④PTZ+SB组(腹腔注射PTZ+侧脑室注射SB);⑤PTZ+SB+OXA组(腹腔注射PTZ+侧脑室注射SB和OXA)。观察各组大鼠的空间学习记忆能力及海马齿状回区BrdU+和BrdU+/NeuN+细胞的表达。结果:与PTZ+OXA组比较,PTZ+SB+OXA组大鼠逃避潜伏期延长、穿越平台象限的次数减少(P<0.05)。免疫荧光显示,PTZ+OXA组大鼠齿状回区BrdU+和BrdU+/NeuN+细胞表达增多(P<0.01),而PTZ+SB+OXA组大鼠齿状回区BrdU+/NeuN+细胞表达比PTZ+OXA组减少(P<0.01)。结论:OXA通过OX1R能改善癫大鼠的空间学习记忆能力,可能与OX1R介导的海马齿状回神经细胞增殖与分化作用有关。     

抑郁状态下大鼠海马Notch1信号系统的改变

目的 了解大鼠抑郁模型中海马神经重塑障碍与Notch1信号系统功能改变的关系.方法 54只大鼠随机分为CUMS 14 d组、CUMS 28 d组和对照组,前两组接受慢性不可预知温和应激和孤养(chronic unpredictable mild stress,CUMS)14 d和28 d建立抑郁模型.采用免疫组化、免疫荧光、RT-PCR和Western blot 法.测定大鼠海马神经干细胞的增殖、存活和分化以及Notch1信号通路各个因子的基因及蛋白表达水平的改变.结果 与对照组比较,CUMS 14 d组和CUMS 28 d组大鼠海马神经干细胞增殖与存活明显减少(P<0.001).CUMS 28 d组大鼠海马神经干细胞分化NeuN/BrdU、GFAP/BrdU比例无明显差异(P>0.05).与对照组比较.CUMS 14 d组和CUMS 28 d组Notch1信号通路各因子(NICD、Hes1、Hes5和Jag1)基因表达和蛋白水平明显降低(P<0.01).结论 抑郁大鼠海马齿状回神经干细胞增殖和存活受到抑制,但分化无改变;同时,大鼠海马Notch1功能下调.提示Notch1信号系统可能与抑郁症海马神经再生障碍有关.     

慢性强迫游泳应激对大鼠海马神经元再生和p-CREB表达的影响

目的 研究慢性强迫游泳应激模型大鼠海马神经元再生和磷酸化环磷酸腺苷反应元件结合蛋白（p-CREB）的表达.方法 30只雄性SD大鼠随机分为3组：强迫游泳7 d组（S1组）、强迫游泳14 d组（S2组）和对照组.S1组和S2组分别连续强迫游泳7 d和14 d,每天5 min,水温（10±0.5）℃.采用免疫组化半定量测定大鼠海马5-溴脱氧尿苷（BrdU）和p-CREB阳性细胞表达情况.结果 免疫组化结果 显示,在整个海马结构中BrdU及p-CREB的阳性细胞主要集中于齿状回的颗粒细胞下层.与对照组比较,S1组、S2组大鼠海马齿状回BrdU和p-CREB阳性细胞数均明显减少（P＜0.01）;而与S1组比较,BrdU阳性细胞数无统计学差异（P＞0.05）,S2组p-CREB阳性细胞数进一步减少（P＜0.01）.结论 慢性强迫游泳应激可导致海马神经元再生功能障碍,其机制可能与p-CREB信号转导通路有关. 基金项目：     

血管性认知障碍小鼠脑内少突胶质细胞再生分化障碍特点研究

目的 观察血管性认知障碍小鼠模型中,缺血性炎性损伤对室管膜下区及海马齿状回少突胶质细胞 再生分化的影响,为血管性认知障碍的缺血性炎症机制提出新的损伤途径。 方法 成年雄性CD1小鼠随机分为模型组和假手术组,每组24只,模型组采用双侧颈动脉反复缺 血再灌注法制备血管性认知障碍小鼠模型。造模后4～6 d连续腹腔注射5 -溴脱氧尿嘧啶核苷 （bromodeoxyuridine,BrdU）（150 mg/kg）标记新生细胞,分别于术后14 d和28 d每组随机取一半小鼠脑 组织进行脑切片免疫组化、免疫荧光双标共聚焦检测,标记脑组织室管膜下区和海马区的少突胶质 细胞、星形胶质细胞及神经元,观察新生少突胶质细胞增殖及分化情况,并观察星形胶质细胞的增 生活化情况。 结果 造模后14 d和28 d室管膜下区新生细胞（BrdU阳性细胞）在模型组较假手术组明显增加（P均 ＜0.001),造模28 d模型组新生神经元（BrdU/NeuN阳性细胞）较假手术组显著增加（P＜0.001)。与假 手术组相比较,术后28 d模型组海马齿状回少突胶质细胞祖细胞显著增多（P＜0.001）;少突胶质细 胞前体细胞显著减少（P =0.006）。造模后28 d模型组海马齿状回新生星形胶质细胞（BrdU/GFAP阳性 细胞）较假手术组显著增加（P =0.015）。 结论 血管性认知障碍小鼠内源性新生细胞增殖区室管膜下区与海马齿状回区均存在新生细胞反 应性增生的情况。新生细胞区分化的主要细胞为星形胶质细胞,而少突胶质细胞分化障碍,可能是血 管性认知障碍患者影像学常见皮层下白质病变的重要原因。     

Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys

To investigate the effect of global cerebral ischemia on brain cell proliferation in young adult macaques, we infused 5-bromo-2'-deoxyuridine (BrdU), a DNA replication indicator, into monkeys subjected to ischemia or sham-operated. Subsequent quantification by BrdU immunohistochemistry revealed a significant postischemic increase in the number of BrdU-labeled cells in the hippocampal dentate gyrus, subventricular zone of the temporal horn of the lateral ventricle, and temporal neocortex. In all animals, 20-40% of the newly generated cells in the dentate gyrus and subventricular zone expressed the neural progenitor cell markers Musashi1 or Nestin. A few BrdU-positive cells in postischemic monkeys were double-stained for markers of neuronal progenitors (class III beta-tubulin, TUC4, doublecortin, or Hu), neurons (NeuN), or glia (S100beta or GFAP). Our results suggest that ischemia activates endogenous neuronal and glial precursors residing in diverse locations of the adult primate central nervous system.     

卡马西平对成年癫大鼠海马齿状回BrdU/NeuN表达水平及其对空间记忆的影响

目的研究卡马西平对成年癫大鼠海马齿状回新生神经元的影响及其与空间记忆之间的关系。方法采用氯化锂和匹罗卡品联合诱导大鼠癫模型,利用5-溴脱氧尿苷嘧啶与神经元核性蛋白双标记观察海马齿状回内源性神经前体细胞分化为成熟神经元的情况;利用行为学分析评价大鼠的空间记忆。结果 (1)卡马西平可增加癫大鼠海马齿状回新生成熟神经元的数量(P<0.05);(2)卡马西平对癫大鼠的空间记忆有明显改善作用(P<0.01)。结论卡马西平增加癫大鼠海马齿状回新生成熟神经元形成,是其改善癫大鼠空间记忆的可能机制之一。     

Enhanced neurogenesis during trimethyltin-induced neurodegeneration in the hippocampus of the adult rat

The occurrence of neurogenesis in the hippocampus of the adult rat during trimethyltin (TMT)-induced neurodegeneration was investigated using bromodeoxyuridine (BrdU). Fifteen days after TMT intoxication, BrdU-labeled cells were significantly more numerous in the hippocampus of treated animals, gradually decreasing towards the control value 21 days after intoxication in the dentate gyrus (DG), while in the CA3/hilus region BrdU-labeled cells were still more numerous in TMT-treated rats. In order to investigate the fate of newly-generated cells double labeling experiments using neuronal or glial markers were performed. Colocalization of the neuronal marker NeuN was detected in many BrdU-positive cells in the DG, while in the CA3/hilus region no colocalization of NeuN and BrdU could be observed. No colocalization of BrdU and the astroglial marker GFAP or the microglial marker OX-42 was detected either in the DG and or in the CA3/hilus region. The results indicate an enhancement of endogenous neurogenesis in the hippocampus during TMT-induced neurodegeneration, with the development of a subpopulation of regenerated cells into neurons in the DG, while in the CA3/hilus region the population of newly-generated cells should be regarded as undifferentiated.     

Increased proliferation of neural progenitor cells but reduced survival of newborn cells in the contralateral hippocampus after focal cerebral ischemia in rats.

Recent studies demonstrated that neurogenesis in the adult hippocampus increased after transient global ischemia; however, the molecular mechanism underlying increased neurogenesis after ischemia remains unclear. The finding that proliferation of progenitor cells occurred at least a week after ischemic insult suggests that the stimulus was not an ischemic insult to progenitor cells. To clarify whether focal ischemia increases the rate of neurogenesis in the remote area, the authors examined the contralateral hemisphere in rats subjected to permanent occlusion of the middle cerebral artery. In the subgranular zone of the hippocampal dentate gyrus, the numbers of bromodeoxyuridine (BrdU)-positive cells increased approximately sixfold 7 days after ischemia. In double immunofluorescence staining, more than 80% of newborn cells expressed Musashi1, a marker of neural stem/progenitor cells, but only approximately 10% of BrdU-positive cells expressed glial fibrillary acidic protein (GFAP), a marker of astrocytes. The number of BrdU-positive cells markedly decreased 28 days after BrdU administration after ischemia, but it was still elevated compared with that of sham-operated rats. In double immunofluorescence staining, 80% of newborn cells expressed NeuN, a marker of differentiated neurons, and 10% of BrdU-positive cells expressed GFAP. However, in the other areas of the contralateral hemisphere including the rostral subventricular zone, the number of BrdU-positive cells remained unchanged. These results showed that focal ischemia stimulated the proliferation of neuronal progenitor cells, but did not support survival of newborn cells in the contralateral hippocampus.     

Temporal changes of neurogenesis in the mouse hippocampus after experimental subarachnoid hemorrhage

Recent studies indicate the existence of progenitor cells and their potential for neurogenesis in the subventricular zone (SVZ) and the hippocampus dentate gyrus (DG) of normal adult mammalian brain. Increased neurogenesis has been shown following cerebral ischemia and traumatic brain injury; however, the involvement of neurogenesis in subarachnoid hemorrhage (SAH) has not been examined. Adult male CD-1 mice were subjected to SAH by endovascular perforation of the left anterior cerebral artery. Mice received intraperitoneal injections of the cell proliferation-specific marker 5'-bromodeoxyuridine (BrdU) after SAH induction. BrdU incorporation was examined from 1 to 30 days after SAH by immunohistochemistry. The BrdU-positive cells were detected in SVZ and DG of normal control brain, and were significantly decreased in both areas three days after SAH. The number of these cells had recovered to its control level seven days after SAH. Double staining with BrdU and NeuN indicated that the majority of the BrdU-positive cells migrating into the granular cell layer of the DG became NeuN-positive 30 days after SAH. In conclusion, temporal changes of the neurogenesis as shown in the present study suggest that neurogenesis in the hippocampus may affect functional outcome after SAH. The induction of the neurogenesis can provide therapeutic value against SAH.     

Temporal profile of neurogenesis in the subventricular zone,dentate gyrus and cerebral cortex following transient focal cerebral ischemia

Abstract

Background: In the adult mammalian brain, it is considered that neurogenesis persists in limited regions such as the hippocampal dentate gyrus (DG) and the subventricular zone (SVZ) of the lateral ventricle. On the other hand, neurogenesis in the cortex after cerebral ischemia and its role in post-stroke recovery have not been clarified yet. In this study, we investigated neurogenesis in the cortex and the spatiotemporal profile of neural progenitors in SVZ and DG of rats subjected to transient focal cerebral ischemia.

Materials and methods: Male Sprague–Dawley rats (270–300 g) were subjected to 60 minute middle cerebral artery occlusion. Proliferating cells were labeled by the cumulative administration of BrdU 1, 2, 3, 4, 6 and 8 weeks after ischemia induction (at weeks 1–4, 6 and 8). Double labeling was also performed with antibodies against BrdU and NeuN.

Results: BrdU-positive cells proliferated in DG and SVZ of the bilateral hemispheres, and their proliferation peaked at week 3 in SVZ and at week 4 in DG. In the peri-infarct zone of cerebral cortex, BrdU-positive cells co-expressed NeuN from weeks 3 to 8.

Conclusion: Neurogenesis was observed in the cerebral cortex and proliferation of neural progenitors occurred in SVZ and DG of rats subjected to transient focal cerebral ischemia. Our data might indicate that endogenous dormant neural stem cells residing in the cortex were activated by ischemic insult to induce the proliferation of neural progenitors and differentiation into mature neurons.     

Temporal changes of neurogenesis in the mouse hippocampus after experimental subarachnoid hemorrhage

Abstract

Recent studies indicate the existence of progenitor cells and their potential for neurogenesis in the subventricular zone (SVZ) and the hippocampus dentate gyrus (DG) of normal adult mammalian brain. Increased neurogenesis has been shown following cerebral ischemia and traumatic brain injury; however, the involvement of neurogenesis in subarachnoid hemorrhage (SAH) has not been examined. Adult male CD-1 mice were subjected to SAH by endovascular perforation of the left anterior cerebral artery. Mice received intraperitoneal injections of the cell proliferation-specific marker 5 ′ -bromodeoxyuridine (BrdU) after SAH induction. BrdU incorporation was examined from 1 to 30 days after SAH by immunohistochemistry. The BrdU-positive cells were detected in SVZ and DG of normal control brain, and were significantly decreased in both areas three days after SAH. The number of these cells had recovered to its control level seven days after SAH. Double staining with BrdU and NeuN indicated that the majority of the BrdU-positive cells migrating into the granular cell layer of the DG became NeuN-positive 30 days after SAH. In conclusion, temporal changes of the neurogenesis as shown in the present study suggest that neurogenesis in the hippocampus may affect functional outcome after SAH. The induction of the neurogenesis can provide therapeutic value against SAH.     

Regeneration of granule neurons after lesioning of hippocampal dentate gyrus: evaluation using adult mice treated with trimethyltin chloride as a model

The hippocampal dentate gyrus in adult animals is known to contain neural progenitors that proliferate and differentiate into neurons in response to brain injury. Little has been observed, however, on regeneration of the granule cell layer of the dentate gyrus that has been directly injured. Using trimethyltin (TMT)-treated mice as an in vivo model, we evaluated the ability of this layer to regenerate after injury. The administration of TMT induced neuronal death in the dentate gyrus selectively 2 days later, with recovery of granule neurons on day 14 and thereafter. At an early stage (days 2-5) after the damage by TMT treatment, 5-bromo-2'-deoxyuridine (BrdU) incorporation into at least two different types of cells was facilitated in the dentate gyrus: BrdU-positive/neuronal nuclear antigen (NeuN)-negative cells were found predominantly in the subgranular zone and granule cell layer, whereas BrdU-positive/NeuN-positive cells were numerous in the dentate molecular layer and hilus. In addition, expression of proliferating cell nuclear antigen, nestin, NeuroD3, and doublecortin, which are markers for proliferating cells and neural progenitors/neuronal precursors, was extremely enhanced in the dentate gyrus at the early stage after treatment. Double staining revealed that BrdU was colocalized with nestin and doublecortin in the subgranular zone. Behavioral analysis revealed that TMT-induced cognition impairment was ameliorated by day 14 after the treatment. Taken together, our data indicate that the hippocampal dentate gyrus itself is capable of regenerating the neuronal cell layer through rapid enhancement of neurogenesis after injury.     

缺血缺氧新生大鼠脑内原位神经干细胞的增殖与分化：可通过针刺任脉、督脉及膀胱经实现？

[摘要] 目的 检测针刺任脉、督脉及膀胱经对新生儿缺血缺氧性脑病模型鼠脑内神经干细胞的影响,分析针刺诱导神经干细胞增殖、分化的机制,为临床针刺治疗新生儿缺血缺氧性脑病提供新的细胞学理论依据。方法 新生7天SD大鼠结扎左侧颈总动脉并缺氧2小时制作新生鼠缺血缺氧性脑病模型。实验动物共分三组：针刺组、对照组和正常组。针刺组每天针刺任脉、督脉及膀胱经治疗一次。对照组及正常组不作针刺处理。各组动物每天两次腹腔注射Brdu用于标记脑内神经干细胞增殖情况。分别于模型建立后3d、7d、14d和28d取脑组织行抗Brdu的免疫组化染色,并于模型建立后40d行免疫荧光双标,分别观察各组动物海马及皮层Brdu阳性细胞数目、形态、分布以及分化情况; 并比较他们之间的差异。结果 抗Brdu的免疫组化染色显示针刺任脉、督脉及膀胱经治疗第3天及第7天时针刺组动物皮层及海马的Brdu阳性细胞数目和对照组相比,差别无统计学意义;针刺治疗第14天及第28天时针刺组动物皮层及海马的Brdu阳性细胞数目明显比对照组多,差别有统计学意义。针刺后第40天免疫荧光双标显示大部分Brdu阳性细胞和神经元标记物NSE共存,少部分和星形胶质细胞标记物GFAP共存。结论 针刺任脉、督脉及膀胱经能促进HIE模型鼠皮层及海马神经干细胞的增殖潜能;针刺任脉、督脉及膀胱经治疗后新生的神经干细胞大部分分化为神经元,提示针刺后新生的神经细胞有可能有效地补充在缺血缺氧中丧失的神经元,并能促进HIE动物功能的恢复。