本文關(guān)鍵詞： 神經(jīng)降壓肽 神經(jīng)病理痛 脊髓前角　出處：《華中科技大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 神經(jīng)降壓肽(neurotensin,NT)是含有13個(gè)氨基酸殘基的腦腸肽,廣泛分布于哺乳動(dòng)物中樞和外周神經(jīng)系統(tǒng)以及胃腸道內(nèi),參與了眾多的生物效應(yīng)。在中樞神經(jīng)系統(tǒng)中研究較多的是其對(duì)痛覺的調(diào)制作用。其中脊髓水平的神經(jīng)降壓肽主要來源于脊髓固有神經(jīng)元內(nèi),具有明顯的鎮(zhèn)痛作用,并推測(cè)該鎮(zhèn)痛作用是由其高親和力受體( NTR1)或低親和力受體(NTR2)介導(dǎo)的。已證實(shí),脊髓前角和后角都含有神經(jīng)降壓肽,對(duì)后角神經(jīng)降壓肽的分布已有 了較詳細(xì)的報(bào)道,而對(duì)其在前角的分布卻知之甚少。另外,參與痛覺調(diào)制的神經(jīng)降壓肽具體來自于脊髓的何種神經(jīng)元亦不明確。本研究首先應(yīng)用形態(tài)學(xué)方法觀察了脊髓前角內(nèi)神經(jīng)降壓肽的分布特點(diǎn),再利用慢性壓迫性坐骨神經(jīng)痛模型觀察脊髓前角和后角的神經(jīng)降壓肽在神經(jīng)病理痛條件下有何變化,從而初步探討了脊髓水平神經(jīng)降壓肽在痛覺調(diào)制中可能的機(jī)制。 一.脊髓前角神經(jīng)降壓肽的分布 本研究首先利用經(jīng)典的尼氏染色法和標(biāo)記乙酰膽堿酯酶的Karnovsky-Roots法來顯示脊髓前角的運(yùn)動(dòng)神經(jīng)元,再利用免疫組織化學(xué)技術(shù)應(yīng)用神經(jīng)降壓肽抗體觀察脊髓前角內(nèi)神經(jīng)降壓肽免疫陽性神經(jīng)元的分布特點(diǎn),通過對(duì)二者的比較,結(jié)果發(fā)現(xiàn): 脊髓前角神經(jīng)降壓肽免疫陽性細(xì)胞可分為大、小兩種。其中大細(xì)胞主要分布在前角前外側(cè)(第Ⅸ層),多為多極神經(jīng)元或雙極神經(jīng)元,與尼氏染色和乙酰膽堿酯酶顯色顯示的脊髓前角運(yùn)動(dòng)神經(jīng)元具有明顯的一致性,可認(rèn)為該類陽性細(xì)胞可能為運(yùn)動(dòng)神經(jīng)元(包括α和γ神經(jīng)元);小陽性細(xì)胞則散在分布于前角各個(gè)區(qū)域,除可能為調(diào)節(jié)性中間神經(jīng)元外,尚不能推測(cè)該類陽性細(xì)胞為何種神經(jīng)元。 二.慢性壓迫性坐骨神經(jīng)痛模型的制備 成年雄性SD大鼠隨機(jī)分為模型組、假手術(shù)組和正常組。其中模型組大鼠通過5-0鉻制羊腸線結(jié)扎單側(cè)坐骨神經(jīng)建立慢性壓迫性坐骨神經(jīng)痛模型,假手術(shù)組大鼠處理除未結(jié)扎神經(jīng)外同模型組,正常組大鼠未做任何處理。采用意大利產(chǎn)UGO-BASILE 37300型輻射熱測(cè)痛儀,以大鼠術(shù)側(cè)后足抬腿潛伏期為其痛閾反映值,連續(xù)監(jiān)測(cè)其痛閾及行為變化。 結(jié)果發(fā)現(xiàn): 模型組大鼠術(shù)后第一天即開始出現(xiàn)術(shù)側(cè)自發(fā)抬足、舔足以及不敢持重等自發(fā)性疼痛和感覺異常現(xiàn)象,痛閾亦隨之降低, 5-7天達(dá)到高峰,伴有足趾屈曲內(nèi)翻等畸形,安靜時(shí)臥于健側(cè),至13-14天上述現(xiàn)象逐漸消失,但足趾屈曲內(nèi)翻以及跛行等畸形無明顯恢復(fù)。假手術(shù)組大鼠無上述典型現(xiàn)象出現(xiàn)。三組大鼠偶有輕微自嗜癥狀。 三.神經(jīng)病理痛條件下脊髓水平神經(jīng)降壓肽含量上調(diào) 正常狀態(tài)下在脊髓水平給予外源性神經(jīng)降壓肽具有明顯的鎮(zhèn)痛效應(yīng)。為探討神經(jīng)病理痛條件下脊髓水平內(nèi)源性神經(jīng)降壓肽是否上調(diào),本研究利用神經(jīng)病理痛模型,應(yīng)用免疫組織化學(xué)技術(shù)等形態(tài)學(xué)方法以及監(jiān)測(cè)大鼠痛閾等行為學(xué)手段,分別在大鼠痛閾最低期(第七天)和恢復(fù)期(第十四天)觀察模型組、假手術(shù)組和正常組大鼠脊髓腰膨大處神經(jīng)降壓肽的含量變化。 結(jié)果發(fā)現(xiàn): 1.在前角前外側(cè)(含第Ⅸ層),模型組大型神經(jīng)降壓肽免疫陽性細(xì)胞數(shù)和陽性面積無論在第7天還是在第14天都與假手術(shù)組和正常組之間沒有顯著性差異,即在痛閾最低期和痛閾恢復(fù)期都沒有明顯變化;與其同一視野的小型陽性細(xì)胞,其細(xì)胞個(gè)數(shù)無論在第7天還是在第14天都與假手術(shù)和正常組之間有顯著性差異(P0.01)即在痛閾最低期明顯增多,至恢復(fù)期有所減少,但均高于假手術(shù)和正常組;在前角內(nèi)側(cè)(約在第Ⅷ層內(nèi)),小型陽性細(xì)胞的細(xì)胞個(gè)數(shù)在第7天與假手術(shù)組和正常組之間有顯著性差異(P0.01),但在第14天三組之間沒有顯著性差異,即在痛閾最低期前角內(nèi)側(cè)小陽性細(xì)胞也明顯增多,至恢復(fù)期降到正常組水平。在脊髓其他部位(包括后角)小型陽性細(xì)胞的數(shù)量在三組之間沒有顯著性差異。 2.模型組大鼠在第7天其脊髓后角膠狀質(zhì)(SG)內(nèi)的神經(jīng)降壓肽含量顯著升高,呈明顯的眉毛狀結(jié)構(gòu),而在恢復(fù)期神經(jīng)降壓肽含量下降,眉毛狀結(jié)構(gòu)隨之消失。在假手術(shù)組大鼠和正常組大鼠沒有觀察到這一現(xiàn)象。 綜上所述,本研究在核團(tuán)水平和細(xì)胞水平觀察了神經(jīng)降壓肽在脊髓前角的分布,初步認(rèn)為脊髓前角運(yùn)動(dòng)神經(jīng)元內(nèi)(包括α和γ神經(jīng)元)含有神經(jīng)降壓肽,另外神經(jīng)降壓肽還存在于前角某些調(diào)制性中間神經(jīng)元內(nèi)。神經(jīng)病理痛條件下,脊髓后角膠狀質(zhì)內(nèi)神經(jīng)降壓肽含量升高,同時(shí)前角含有神經(jīng)降壓肽的中間神經(jīng)元數(shù)量上調(diào),二者在形態(tài)學(xué)和行為學(xué)上具有一致性,因此可推斷脊髓水平的神經(jīng)降壓肽在后角膠狀質(zhì)內(nèi)參與了痛覺的調(diào)制,而該神經(jīng)降壓肽可能主要來自脊髓前角某些中間神經(jīng)元內(nèi)。
[Abstract]:Neurotensin (neurotensin, NT) is a brain gut peptide containing 13 amino acid residues, is widely distributed in mammalian central and peripheral nervous system and gastrointestinal tract, involved in numerous biological effects. More research in the central nervous system is made of pain. The spinal cord nerve antihypertensive peptides mainly derived from intrinsic neurons of spinal cord, has obvious analgesic effect, and that the analgesic effect is determined by its high affinity receptor (NTR1) or low affinity receptor (NTR2) mediated. Confirmed that the anterior horn of the spinal cord and the angle containing neurotensin, the distribution of neurotensin angle the
A detailed report, and its distribution in the anterior horn is poorly understood. In addition, the neurotensin neurons involved in pain modulation in the spinal cord from the specific is not clear. In this study we used morphological method to observe the distribution of the angle of neurotensin in the spinal cord, with chronic constrictive injury the observation model of neurotensin in spinal cord anterior horn and posterior horn of any changes in neuropathic pain conditions, so as to explore the probable mechanism of the spinal cord level of neurotensin in pain modulation.
The distribution of neurotensin in the anterior horn of the spinal cord
The Karnovsky-Roots method in the study of classical Nissl staining and acetylcholinesterase markers to show motor neurons in anterior horn of spinal cord, the distribution of neurotensin immunoreactive neurons in spinal cord anterior horn were observed by immunohistochemical technique of neurotensin antibody, through the comparison of the two results:
The anterior horn of spinal cord of neurotensin immunoreactive cells can be divided into large, small two. The cells were mainly distributed in the anterior horn of the anterolateral (Ninth layer), multi multipolar neurons or bipolar neurons and Nissl staining and acetylcholinesterase staining showed spinal motor neuron has obvious consistency, can as the cells into motor neurons (including alpha and gamma neurons); small positive cells scattered in the anterior horn of all regions, except possibly for the regulation of interneurons, still can not speculate the positive cells what kind of neurons.
Two. Preparation of chronic compression sciatica model
Adult male SD rats were randomly divided into model group, sham group and normal group. The rats in the model group by 5-0 chromic catgut ligation of unilateral sciatic nerve to establish chronic constrictive injury model, the rats in the sham operation group without nerve ligation treatment with the model group, the rats in the normal group without any treatment. The Italy UGO-BASILE 37300 radiation thermal dolorimeter, rat ipsilateral foot leg latency as the pain threshold value of continuous monitoring, the pain threshold and behavioral changes.
The results were as follows:
The rats in the model group after the first day of the operation side began to appear spontaneous lifting, abnormal phenomenon and not so serious licking spontaneous pain and feel pain also decreased, reaching a peak at 5-7 days, accompanied by toe flexion varus deformity and so on, quiet when lying on the contralateral side, to 13-14 days the phenomenon gradually disappear, but toe buckling and varus deformity had no obvious limp recovery. The rats in the sham operation group without the typical phenomenon. The three groups of rats from occasional minor symptoms.
Three. Up regulation of the level of neurotensin in the spinal cord under the condition of neuropathic pain
Under normal conditions the exogenous neurotensin has a significant analgesic effect given at the spinal level. To investigate neuropathic pain conditions of spinal cord levels of endogenous neurotensin is up-regulated, the neuropathic pain model, by means of immunohistochemistry and morphological methods and monitoring the pain threshold behavior in rats, respectively. The lowest threshold period (seventh days) and recovery period (Fourteenth days) to observe the content changes of the model group, sham operation group and normal group rat spinal cord lumbar enlargement of neurotensin.
The results were as follows:
1. in the anterior horn of anterolateral (including ninth layer), the model group of large neurotensin immunoreactive cell number and the positive area in seventh days or in Fourteenth days and between sham operation group and the normal group had no significant difference, which is in the lowest pain threshold period and the recovery of the pain threshold did not change significantly; small positive cells with the same vision, the number of cells in seventh days or fourteenth days in between the sham and normal groups had significant difference (P0.01) significantly increased in the lowest threshold period, recovery period has decreased, but were higher than those in the sham and normal groups; in the anterior horn medial (about Article VIII layer), there was significant difference between the seventh day and the sham operation group and the normal group number of cells with a small positive cells (P0.01), but no significant difference in the fourteenth days between the three groups, which was significantly increased in the minimum period before the small angle threshold to restore positive cells. The number of small positive cells in the other parts of the spinal cord (including the posterior horn) was not significantly different between the three groups.
2. rats in the model group on the seventh day of the spinal dorsal horn of substantia gelatinosa (SG) neurotensin content increased significantly, showing the apparent eyebrow like structure, while the decline in the period of neurotensin content recovery, eyebrow like structure disappears. In the sham operation group rats and normal rats did not observe this a phenomenon.
In summary, this study investigated the distribution of neurotensin in the spinal ventral horn in the nuclei and cellular levels, primarily that spinal cord motoneurons (including alpha and gamma neurons) containing neurotensin, and neurotensin also exist in anterior horn neurons. Some intermediate modulation of neuropathic pain conditions neurotensin content increased, horncinerea in spinal cord neurons increased at the same time, the middle anterior horn containing neurotensin, two are consistent in morphology and behavior, so it can be concluded that neurotensin levels of the spinal cord involved in pain modulation in the posterior horncinerea, the neurotensin may mainly come from some intermediate neurons in the anterior horn of the spinal cord.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R363

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