大鼠脑内γ-干扰素样免疫反应产物分布及电针后变化

【作者】 孙志宏；

【导师】 欧阳五庆； 李育良；

【作者基本信息】 西北农林科技大学 ， 基础兽医学， 2001， 硕士

【摘要】 研究大鼠脑内γ-干扰素（IFN-γ）样免疫反应产物的分布及其电针刺激足三里穴后的变化，为免疫-神经-内分泌调节网络学说增添形态学证据，也为阐明针灸作用机理提供形态学资料。将健康青年SD大鼠随机分为四组：正常对照组，免疫抑制对照组，电针组，免疫抑制+电针组，采用免疫组织化学SP法对大鼠大脑皮层、海马、室管膜、下丘脑、小脑、延髓内脏带进行了研究，结果显示： 1．大脑皮层 大鼠大脑皮层内均可见IFN-γ样免疫反应产物，但各层分布不均，Ⅰ、Ⅱ、Ⅳ和Ⅵ层较少，Ⅲ和Ⅴ层较多。与正常对照组大鼠相比，电针刺激大鼠足三里穴对大脑皮层内IFN-γ样免疫反应阳性细胞数量无明显影响；但与免疫抑制对照组大鼠相比，电针刺激免疫抑制大鼠足三里穴，大脑皮层Ⅲ和Ⅴ层内IFN-γ样免疫反应阳性细胞数量明显增多，而Ⅰ、Ⅱ、Ⅳ和Ⅵ层数量增多，但无统计学意义（以运动区作为统计对象）。推测IFN-γ在大脑皮层具有免疫调节功能，电针刺激免疫抑制大鼠足三里穴可以提高大脑皮层的免疫调节功能。 2．海马 大鼠海马CA1、CA2、CA3区均存在IFN-γ样免疫反应产物，但各区阳性细胞数量所占比例不同，比例最高的是CA1区，多分布于锥体细胞层，其余二区内IFN-γ样免疫反应阳性细胞数量较少。此外，在灰被、内也可见数量较少IFN-γ样免疫反应阳性细胞。与正常对照组大鼠相比，电针刺激大鼠足三里穴对海马内IFN-γ样免疫反应阳性细胞数量无明显影响；但与免疫抑制对照组大鼠相比，电针刺激免疫抑制大鼠足三里穴，海马针刺效应最明显的部位是CA1区，IFN-γ样免疫反应阳性细胞数量有明显增多，其余二区和灰被有变化，但不明显，无统计学意义。推测海马除具有学习和记忆功能外，还具有免疫调节功能。IFN-γ可能参与海马的免疫调节，电针刺激免疫抑制大鼠足三里穴可以提高海马的免疫调节功能。 3．室管膜 大鼠室管膜侧脑室、第三脑室、中脑导水管和第四脑室壁上均存在IFN-γ样免疫反应阳性细胞。与正常对照组大鼠相比，电针刺激大鼠足三里穴对空管膜壁上IFN-γ样免疫反应阳性细胞数量无明显影响；但与免疫抑制对照组大鼠相比，电针刺激免疫抑制大鼠足三里穴仅在中脑导水管壁上IFN-γ样免疫反应阳性细胞数量有明显增多，而侧脑室、第三脑室和第四脑室壁上IFN-γ样免疫反应阳性细胞数量均未见有明显增多。推测大鼠室管膜可能具有合成或摄取IFN-γ的能力，室管膜内的IFN-γ样免疫反应阳性细胞可能与脑-脑脊液之间的信息传递有关。电针刺激免疫抑制大鼠足三里穴可使室管膜细胞合成或摄取IFN-γ的能力得到提高。 4．下丘脑 大鼠下丘脑室旁核、室周核、视上核、视上交叉核、乳头体后核等部位均存在IFN-γ样免疫反应阳性细胞。与正常对照组大鼠相比，电针刺激大鼠足三里穴对下丘脑诸核内IFN-γ样免疫反应阳性细胞数量无明显影响；但与免疫抑制对照组大鼠相比，电针刺激免疫抑制大鼠足三里穴，下丘脑室旁核、室周核内IFN-γ样免疫反应阳性细胞数量有明显增多，而视上交叉核、视上核、乳头体后核无明显变化。推测IFN-γ可能参与下丘脑的免疫调节，电针刺激免疫抑制大鼠足三里穴可提高下丘脑的免疫调节功能。 5．小脑 大鼠小脑皮层分子层内存在大量IFN-γ阳性纤维，偶见分散的阳性细胞，而浦肯野氏层存在大量IFN-γ样免疫反应阳性细胞，颗粒层阳性细胞呈散在分布。与正常对照组大鼠 相比，电针刺激大鼠足三里穴对小脑皮层内IFNq样兔疫反应阳性细胞数量无明显影响；但与 兔疫抑制对照组大鼠相比，电针刺激兔疫抑制大鼠足三里穴，小脑浦肯野氏层内IFN－丫样免疫 反应阳性细胞数量明显增多，分子层和颗粒层无明显变化。 大鼠小脑顶核、间位核和齿状核内均存在IFNff样免疫反应阳性细胞，其中顶核内阳性细 胞数且较少，分布较稀疏，而间位核和齿状核内存在较多阳性细胞。与正常对照组大鼠相比， 电针刺激大鼠足三里穴对小脑核内IFNq样免疫反应阳性细胞数诅无明显彤响；但与免疫抑制 对照组大鼠相比，电针刺激免疫抑制大鼠足三里穴，间位核和齿状核内IFNff样免疫反应阳性 细胞数量明显增多，而顶核无明显影晌。 提示小脑除具有运动、学习和记忆功能外，可能还具有免疫调节功能。电针刺檄免疫抑制 大鼠足三里穴可以提高小脑的免疫调节功能． 6．延髓内脏带 大鼠延们内脏带的迷走补经背核、孤束核、疑核、腹外恻网状核、旁正中网 状核等核内均存在较多IFNq样免疫反应阳性细胞，其中疑核、腹外例网状核、旁正中网状核 内胞质着色较深，途走神经背核和孤束核内胞质着色戏浅．与正常对照组大鼠相比，电针刺激 大以足三里穴对延历内脏带内IPNq样免疫反应阳性细胞数量无明显影叼：但与免疫抑制对照 组大鼠相比，电针刺檄免疫抑制大鼠足三里穴，疑核、腹外侧囚状核、旁正中囚状核内IFNq 样免疫反应阳?更多还原

【Abstract】 Tar DISTareUTION AND CRANGE OF ELECTROACUPUNCTU’RE ON TIIEIFNry LIus IMMUNOREAETIVE PRODUTION IN RAT BRAINANSTAnCTThe exPression of IFN- y like immunoreactivity(IFN- Y-li) in rat brain was stUdied and thecorrelation betWeen acuPunCtUre and immuno-neuro-endocrine net was discussed. The rat weregrouped to contrOl group, Zusanli POint grouP, immunosuppresive group and immunosupPtessive+electroacupuntUre grouP. The ZuSanli point of rat was electrOacupuntUre by using acupunctUreinstrUment (G6805-2A, intensity was 3V frequent was 2HZ, time was 30 minute), and the changes ofthe distribUtion of IFN- Y -li in cerebral corteX, hippocaInPus, ePendyIn4 hyPOthalamus, CerebeIlum,medullny viscetal zone were stodied by using the technique of inununohistOchemiStry SR The resultSas fOllow;:l. Cerebral cortexThe IFN- Y -li neurons were abundanly distributed in all layer of cerebra cortex, but especiallywere distributed in the lnyers of III. V. Compare with control group, The number of IPN- Y -lineurons in electroacuPwte at Zusanli point had no significan difference. The IFN- Y -li neuronshad significant difference in lagers of llI. V, when compared wun immunosuPpressive grouP. It isrecreasing in lagers of I 1 II’ W’ V,bul there is not significant in statiStics. The resultS showed thatIFN- Y existCd in cerebral cortCx of tat, and IFN- y may play the role of immunomodulation in cerebralcortex.2. HipPOcampusThe lFN- Y -li positive neurons extensively distributed in CAl’ CA2. CA3 of hippocampus incontrOl group, especially in CAl, and more positive neurons were in the pyramidal cell layers of thehippocampus .In few positive neurons waS deteCted in CA2. CA3. The effect of EA on IFN- Y -lipositive productS in hippocampus wasn’t obvious when compared with control group. The IPN- Y -lipositive cells were extensively detected in CAl of hippocampus when compared withimmunosuppressive group.The resultS suggested that the IFN- y may play the role ofimmunomodulation in hippocampus.3. EpendymaThe IFN- Y -li POsitive cells were distribUted in LV V3, Aq, V4. ComPared with cchl grouP, thereis no signican in electrOacupunbe group. The change Of IFN- Y -like positive cells were not obviousin Zusanli group when comPared with control group. The Iru- Y -li POsihve cells in AqcimmunosuPpressivatEA grOuP much than immunosuPPressive grOup. The resultS suggsted that the IFN-y can Synthnhe or absorb in ependyIna and the immunsuPPressive +EA grouP can lead tO occur IFN-?IFN- y may be a novel neuopeptide in epen dyma and modulate brain-CSF circuit.4. HypothalamusThe IFN- i -ii positive cell were detected in nucleus paraventricularis, nucleus periventricularis, nucleus supra-opticus, nucleus suprachiasmaticus, nucleus mamillaris posterior Compared with control group, electroacupunture group bad no signicant difference in the number of the IFN- Y -Ii positive cells in hypothalanus. The effect of inimunosuppressive+EA group on IFN- y -Ii positive cells were obvious in nucleus paraventricularis, nucleus periventricularis when compared with cimmunosuppressive group. The result showed that the hypothalamus have inmunomodulation and the immunosuppressive+EA group can lead to occur IFN- Y.5. CerebellumThe IFN- 慳?positive fibers were widely distributed in stratum moleculare of cerebellum , The IFN慳?-ii positive cell were weekly occasionally detected in those. The IFN- 慳?-li positive cell were widely detected in layers of Purkinje and weeklyin the stratum graulosum. Electroacupunture group had no signicant difference in the number of the IFN- 慳?-ii positive cells when compared with control group.The IFN- 慳 -Ii positive cells were obvious in stratum Purkinje, when compared with cimmunosuppressive group. The result suggested that IFN- 慳?could have imniun抋o modulate action in cerebellum.The IFN- 慳 -Ii positive neurons extensively distributed in nucleus fastigi, nucleus interpostitus, nucleus dentatus of cerebellum. The effect of EA on IFN- 慳?-Ii posi更多还原