【作者】 邢旭光；

【导师】 沈伟哉；

【作者基本信息】 暨南大学 ， 人体解剖学， 2002， 硕士

【摘要】 目的： 观察脑缺血大鼠脑内部分区域nNOS和IGF-Ⅱ免疫阳性神经元的变化和皮质神经元超微结构变化以及复方丹参注射液的保护作用。 方法： SD大鼠72只，成模后随机分为4组：(1)假手术组；(2)2VO组；(3)2VO+NS组；(4)2VO+DS组；每组18只。分别于手术后8、16、24周各组分别取相同例数大鼠(n=6)处死取材。ABC免疫细胞化学方法染色方法显示nNOS和IGF-Ⅱ免疫阳性神经元，并作定量分析，同时用透射电镜技术观察大脑皮质神经元超微结构的变化，以及复方丹参对脑缺血后损伤的影响。 结果： (1) 缺血8周时2VO和2VO+NS组Acb内nNOS阳性神经元数目明显降低，脑缺血16周时虽有升高但仍明显低于假手术组并有显著差异(P＜0.05)，到脑缺血24周时恢复正常水平：整个实验期间2VO+DS组Acb内nNOS阳性神经元数目始终高于前两组，并与假手术组无显著差异。 (2) 缺血8周时2VO、2VO+NS和2VO+DS组S1内nNOS阳性神经元数目明显升高并与假手术组有显著差异(P＜0.05)：在脑缺血16周时2VO和2VO+NS组S1内nNOS阳性神经元数目恢复正常水平，2VO+DS组Acb内nNOS阳性神经元数目稍高于假手术组但无显著差异。 (3) 整个实验期间，假手术组PVN内nNOS阳性神经元数目无明显改变。从脑缺血后8周到16周和24周时，2VO、2VO+NS和2VO+DS组PVN内nNOS阳性神经元数目明显升高且无变化，与假手术组相比有显著差异(P＜0.05)。 (4) 缺血8周时2VO和2VO+NS组大鼠脑皮质IGF-Ⅱ阳性神经元变化明显增多，到脑缺血16和24周时逐渐下降，24周时仍多于假手术组。同时期2VO+DS组神经元均较2VO和2VO+NS组减少，但仍较假手术组增多。 暨南大学硕士论文 脑缺血大鼠脑 nNOS和 IGF－11阳性神经元的变化及复方丹参的保护作用 O）缺血8周时ZVO组大脑皮质内神经元核溶解，胞质内细胞器溶解减少；到脑缺 血16周时细胞结构基本完整，但仍有核膜内陷、染色质边集、细胞器不清等表现； 到脑缺血24周时基本恢复正常。同时期ZVO＋DS组神经元均较ZVO组好转。 结论： ＊）脑缺血后Acb内nNOS阳性神经元先减少后回复至正常，提示缺血后Acb内 NO的功能与其它脑区不同，在脑缺血早期其功能可能受到抑制。 口）脑缺血后 SI区 nNOS阳性神经元先增多后减少至正常水平，提示缺血后早期 皮质内nNOS阳性神经元增多可能是缺血刺激后的应激性增多；而缺血中晚期皮质 nNOS阳性神经元减少可能与脑缺血后期神经元的大量丢失有关。 ＊）脑缺血后PVN内nNOS阳性神经元持续增多，这一变化提示脑缺血后下丘脑 PVN内NO的功能活动增强，NO可能影响下丘脑AVP和OT等激素的分泌，以 及具有增强交感神经活性的作用。 （）脑缺血后皮质IGF刁阳性神经元数量显著升高，并随脑损伤的修复进程而减 少，提示IGF－11可能是一种参与脑缺血急性期的抗损伤因子，但在脑缺血晚期合成 的量较少。 ③ 复方丹参在脑缺血早期对神经元有较好的保护作用，其机制可能是通过调节 NO和IGF等细胞因于的量而完成的。更多还原

【Abstract】 PURPOSE:Our experiment aim to the discusses the protection mechanism in the development of cerebral ischemia by observing the changes of neuronal nitric oxide synthase (nNOS) and insulin-like growth factor- II (IGF- II ) immunocytochemical reactivity neurons in some cerebral nucleus, the ultrastructure changing of neuron in cerebral cortex and the effect of used Salvia miltiorrhiza compound.METHOD:12 SD rats were divided randomly into 4 groups, each contained 18 rats. The control group was only separate the bilateral carotid artery. The other 3 groups were first established bilateral carotid artery ligation (2VO) cerebral ischemia rats, and then were separately with 2VO group, 2VO+NS group and 2VO+DS group. At the end of 8, 16 and 24 weeks, the equal number of rats in every group was sacrificed (n=6 each time). The number of nNOS positive neurons in Acb, SI and PVN were counted and then quantitatively analyzed. Qualitative analysis of IGF- II positive neurons and ultrastructure changing in the cerebrum cortex neurons.RESULTS:(1) The number of nNOS positive neurons in Acb of 2VO and 2VO+NS groups significantly decreased at 8 weeks after ischemia; At ischemia 16 weeks still remain low level and have the significant difference compare to the control group, but till ischemia 24 weeks it have raised to the normal level; in the whole course of ischemia (within 24 weeks), the number of nNOS positive neurons of 2VO+DS groups have no changing but always higher to the 2VO and 2VO+NS groups, and have the significant difference compare to the control group.(2) The number of nNOS positive neurons in S 1 of all 3 ischemia group ( 2VO, 2VO+NS and 2VO+DS group) get to a higher level in ischemia 8 weeks; But decreased to the normal level in ischemia 16weeks, although the number of nNOS positive neurons in SI of 2VO+DS group significantly still higher to the control group, but there have no significant difference.(3) In the whole experiment period, the number of nNOS positive neurons in PVN of the control group has no changing. From Sweeks to 24weeks after ischemia, the number of nNOS positive neurons in PVN of the 2VO, 2VO+NS and 2VO+DS groups increasing significantly, and have no significant difference in each other. But compare to the control group, there have significant difference. And have no changing with developing of ischemia.(4) The IGF- II positive neurons number in the cerebrum cortex of 2VO and 2VO+DS groups increased significantly in ischemia 8 weeks, then decreased gradually but not to normal level till ischemia 6month. But in all course of ischemia (within 6 months), the IGF- II positive neurons number in the cerebrum cortex of 2VO+DS group is always lower to the 2VO group at the same time point.(5) In 2VO group after ischemia 8 weeks, the ultrastructure changing in the cerebrum cortex neurons showed karyolysis, organelle dissolve decrease in cytoplast; At ischemia 16 weeks, the neuron structure have removed to integrated, but still have the ischemia exhibition such as karyotheca invagination, chromatin side aggregation, organelle dis-complete. Up to ischemia 24 weeks, the neurons have recovered to the normal structure. And in all course of ischemia (within 6 months), the cell structure of 2VO+DS group is always better to the 2VO group at the same time point.CONCLUSION:(1) The nNOS positive neurons number of Acb is reduce after ischemia but then come round to the normal level in the later, that demonstrated the function of NO in Acb isdifferent to the other brain section and may be restrained in earlier stage of ischmia.(2) The change of nNOS positive neurons number in S1 showed that the nNOS positive neurons increasing in earlier stage of ischmia is excitability increasing. And the reducing in the later is related to the neurons death in the ischemia middle and late stage.(3) The number of nNOS positive neurons in PVN increased significantly in the whole period of ischemia, which implied that the changes of nNOS activity in PVN might be related to the change更多还原