大鼠中枢神经系统对脾脏神经支配的调控——假狂犬病毒跨突触标记

The Pathway and Mechanism Whereby the Central Nervous System Regulates the Innervation of the Spleen in the Rat——Pseudorabies Virus Transsynaptic Tracing Technique

为探讨大鼠中枢神经系统与脾脏之间的神经通路联系及大鼠中枢神经系统对脾脏免疫功能的调控机制 ,采用免疫组化法及荧光金 (Flouro- Gold,FG)神经追踪法研究了假狂犬病毒 (PRV )及 FG注入脾脏 72小时、96小时后逆行标记的 FG神经元在腹腔神经节内、跨突触标记的 PRV神经元及神经纤维在脾脏及神经系统内的分布。结果 :1各时间组大鼠脾被膜及实质内均见 PRV标记的呈串珠状的神经纤维 ,有的纤维以终末与免疫细胞密切接触。 2各时间组大鼠下胸段 (T1 0～ 1 2 )脊神经节实质内见 PRV标记阳性神经元 ,被膜内及实质的神经元之间可见 PRV阳性神经纤维。 3各时间组大鼠腹腔神经节内见散在的 FG标记神经元。 4各时间组大鼠中胸段(T5～ 9)脊髓侧角中间外侧柱 ,后角背侧固有核及胶状质 ,延髓迷走神经背核和孤束核内均见 PRV标记阳性神经元胞体及突起。5 96小时组大鼠除上述阳性结果外 ,标记细胞还可见于下丘脑室旁核。由此推测神经系统与脾脏之间可能存在下列神经通路联系 :1下丘脑室旁核→延髓迷走神经背核→脊髓侧角中间外侧柱的交感神经节前神经元→交感神经腹腔节神经元→脾脏。2脾内感觉神经纤维→脊神经节感觉神经元→脊髓后角背侧固有核和胶状质联络神经元→延髓孤束核神经元→下丘脑感觉中枢。

This investigation was made with scientific inquiry about the neural pathway connection between the central nervous system and the spleen, and the mechanism whereby central nervous system regulates the spleen. Pseudorabies virus (PRV) transsynaptic labeling method, flouro gold (FG) retrograde tracing and immunohistochemical techniques were used. 8 male SD rats were divided into 3 groups (4 for 72h and 2 for 96h with PRV, 2 for 72h with FG).The animals received splenic injection of PRV and FG. After 72h's and 96h's survival, the animals were perfused with fixative containing 4% parafomaldehyde and 0.2% picric acid (pH7.4). Blocks were cut into 40μm sections on cryostat. The sections were handled with ABC immunohistochemical method. The results showed: ①The PRV labeled fibers were found in the capsule and peripheral artery lymph sheath, in the latter the fibers surrounded the blood vessels and located among the lymphocytes of the white and red pulp. Some terminals were in tight contact with splenocytes in each timed group. ②In lower thoracic (T 10~12 ) dorsal root ganglia (DRG) of each timed group, PRV positive neurons were found, and some PRV positive fibers were distributed in the capsule and among the neurons. ③The FG positive neurons were found in the celiac ganglion in each timed group.④ In the intermediolateral nucleus of lateral horn and nucleus of dorsal horn of spinal cord , and in the dorsal motor nucleus of vague (DMNV) and nucleus of solitary tract in medulla, some PRV positive neurons were found in each timed group.⑤In the 96h group, PRV positive neurons were also found in the paraventricular nucleus of the hypothalamus (PVN). From these results we conclude that two possible neural pathways between nervous system and spleen may exist: ① PVN→DMNV→sympathetic preganglia neurons of lateral horn in spinal cord→postganglia neurons of celiac ganglia→spleen.②Sensory terminals of spleen → sensory neurons of DRG → neurons in dorsal horn of spinal cord → nucleus of solitary tract → higher sensory central in PVN.