發(fā)布時間：2018-05-17 07:36

  本文選題：腰椎間盤突出 + 軟骨細胞簇�。� 參考：《天津醫(yī)科大學》2016年碩士論文

【摘要】：第一部分LDH患者椎間盤組織中炎性相關(guān)因子表達的一般特點目的:明確不同類型腰椎間盤突出癥(LDH)患者間盤組織的病理學特點,歸納出炎性相關(guān)因子在間盤組織中表達分布情況,探討間盤組織中炎性因子的來源及其可能病理生理學意義。方法:選取天津醫(yī)院脊柱外科于2014年12月到2015年9月所收治、確診并接受手術(shù)治療的LDH患者26例。根據(jù)臨床癥狀體征、影像學檢查(CT、MRI)結(jié)果及術(shù)中所見纖維環(huán)的完整性將所選病例分為兩組:(1)損傷疝出型椎間盤突出組(R組)(2)退變突出型椎間盤突出組(NR組)。采用HE染色觀察不同分型間盤組織的病理學特點,通過免疫組化、Westernblot等方法檢測IL-17、IL-23、NLRP3、Caspase-1等炎性相關(guān)因子在間盤組織中的表達分布情況,進而歸納出炎性相關(guān)因子在組織中表達的一般特點。結(jié)果:兩種類型LDH患者的間盤組織均表現(xiàn)出退變征象,但損傷疝出組病理表現(xiàn)更為復雜、退變更加明顯,主要表現(xiàn)為:組織裂隙明顯增多,纖維排列更加紊亂、疏松,甚至斷裂;組織邊緣�？梢娎w維素樣壞死、粘液樣變性、滑膜樣化生、鈣結(jié)晶沉積、新生血管長入、炎性細胞浸潤和軟骨細胞簇形成等現(xiàn)象。其中細胞增生和軟骨細胞簇形成部位間盤退變也較為明顯。免疫組化可知IL-17、IL-23、NLRP3、Caspase-1等在兩種類型的間盤組織中均有表達,且損傷疝出型表達含量高于退變突出型;各炎性相關(guān)因子在軟骨樣的髓核細胞中均可見陽性表達,且在組織裂隙周圍的髓核細胞中、軟骨細胞簇中、浸潤的炎性細胞周圍及新生血管周圍陽性表達結(jié)果尤為明顯。結(jié)論:1.兩種類型LDH患者的間盤組織病理學差異較為明顯,提示可能有更多的機制參與間盤組織損傷疝出后的病理表現(xiàn)。增生細胞和軟骨細胞簇部位組織退變程度明顯,提示軟骨細胞簇形成在間盤病理變化中可能發(fā)揮著一定的作用。2.間盤細胞能夠分泌炎性相關(guān)因子,且在間盤細胞增生處和軟骨細胞簇中表達呈強陽性,提示間盤細胞可能與浸潤的免疫細胞一起在間盤突出后誘發(fā)的炎癥過程中發(fā)揮著一定的作用。第二部分體外培養(yǎng)人髓核細胞炎性相關(guān)因子分泌特性的實驗研究目的:腰腿疼痛是間盤源性相關(guān)疾病的主要癥狀,炎癥作為疼痛的主要誘因,在其中可能發(fā)揮著重要的作用。近來研究發(fā)現(xiàn)炎癥的主要介質(zhì)炎性因子在病變的間盤中表達升高,支持上述假說,但其來源尚有一定的爭論,人間盤細胞是否能分泌炎性因子及其分泌特點尚鮮有研究報道,本部分擬對此進行初步的探究。方法:取退變突出型LDH患者術(shù)中摘除的間盤組織,仔細分離培養(yǎng)后獲得髓核細胞,采用P2代進行后續(xù)實驗研究。采用番紅O、甲苯胺藍、瑞士姬姆薩染色及Ⅱ型膠原免疫組化染色等技術(shù)對髓核細胞進行鑒定。對髓核細胞進行饑餓處理或脂多糖(LPS)刺激一段時間后(4h、16h、24h、48h),采用RT-PCR法檢測炎性相關(guān)因子IL-1β、IL-12、IL-17、IL-23、TNF-α、EBI3、IL-37的表達情況,同時采用ELISA方法對髓核細胞培養(yǎng)液中IL-1β、IL-17、TNF-α的蛋白含量進行檢測驗證。結(jié)果:經(jīng)鑒定所培養(yǎng)細胞為髓核細胞,通過PCR發(fā)現(xiàn)基礎狀態(tài)下髓核細胞中不僅有IL-1β、IL-12、IL-17、IL-23、TNF-α等炎性因子的表達,也有抗炎因子EBI3(IL-35)、IL-37的表達,且經(jīng)饑餓或LPS刺激后其表達量較基礎狀態(tài)下明顯增高。采用ELISA方法從蛋白角度對IL-1β、IL-17、TNF-α在細胞培養(yǎng)液的含量進行檢測,也呈現(xiàn)出相似的表達特點。結(jié)論:人髓核細胞不僅能夠表達多種炎性因子,也能夠表達抗炎因子。在饑餓或LPS刺激后其表達量升高,且呈一定的時間依賴性。由于這些炎性相關(guān)因子在間盤基質(zhì)合成與分解、血管生成、神經(jīng)內(nèi)生長、炎癥進展、疼痛致敏等方面發(fā)揮著重要的作用,對這些炎性相關(guān)因子分泌及作用機制的認識有助于我們了解間盤的生物學特性并為干預或治療間盤相關(guān)疾病提供提供新的思路和理論依據(jù)。
[Abstract]:Part 1 the general characteristics of the expression of inflammatory factors in the intervertebral disc tissue of LDH patients: to clarify the pathological characteristics of intervertebral disc tissue in different types of lumbar intervertebral disc herniation (LDH), and to summarize the distribution of inflammatory factors in the intervertebral disc tissue, and explore the source of inflammatory factors in the intervertebral disc and their possible pathophysiology. Methods: 26 patients with LDH in Tianjin Hospital from December 2014 to September 2015 were selected and treated with surgical treatment. According to the symptoms and signs, the results of CT (MRI) and the integrity of the fibrous ring during the operation, the selected cases were divided into two groups: (1) the degeneration of herniated herniation group (group R) (2) degeneration. The protruding intervertebral disc herniation group (group NR). The pathological features of different types of intervertebral disc tissues were observed by HE staining. The expression and distribution of inflammatory related factors such as IL-17, IL-23, NLRP3 and Caspase-1 in the intervertebral disc were detected by immunohistochemistry and Westernblot, and the general characteristics of the inflammatory related factors in the tissue were summarized. Results: the intervertebral disc tissues of the two types of LDH patients showed signs of degeneration, but the pathological features of the injured hernia group were more complex, and the degeneration was more obvious. The main manifestations were: the tissue fissure increased obviously, the fiber arrangement was more disorder, loosely and even broken, and the fibrous necrosis, mucoid degeneration, synovial metaplasia and calcium crystallization were often seen on the edge of the tissue. Deposition, neovascularization, inflammatory cell infiltration, and chondrocyte cluster formation, among which cell proliferation and cartilage cell cluster forming part of disc degeneration are also obvious. IL-17, IL-23, NLRP3, Caspase-1, and other two types of intervertebral disc tissues are observed by immunohistochemistry, and the expression content of the damaged hernia type is higher than that of the degenerative protruding type. All the inflammatory factors were positive in the chondroid nucleus pulposus cells, and in the nucleus pulposus cells around the tissue fissure, the positive expression of the infiltrating inflammatory cells and around the neovascularization was particularly obvious in the chondrocyte clusters. Conclusion: the pathological changes in the intervertebral disc of the 1. types of LDH patients were more obvious, suggesting that the intervertebral disc of the two types of patients were more obvious. There are more mechanisms to participate in the histopathological manifestations of intervertebral tissue injury after hernia. The degree of degeneration is obvious in the tissue of proliferative and chondrocytes, suggesting that the formation of chondrocyte clusters may play a role in the pathological changes of intervertebral disc. The.2. intervertebral disc cells can secrete inflammatory factors, and at the proliferation and cartilage of the intervertebral disc cells. The expression in the cluster is strongly positive, suggesting that the intervertebral disc cells may play a role in the inflammatory process induced by the intervertebral disc herniation with the infiltrating immune cells. Second the experimental study on the secretion characteristics of the inflammatory related factors in human medullary cells in vitro As the main cause of pain, it may play an important role in it. Recent studies have found that inflammatory factors in the main mediators of inflammation are expressed in the disks, which support the hypothesis, but there is still a debate on whether the source of human disc cells can secrete inflammatory factors and their secretions. Methods: the intervertebral disc tissue extirpated in the LDH patients with degenerative protrusion was taken, the nucleus pulposus cells were obtained after careful separation and culture, and the P2 generation was used for subsequent experimental study. The nucleus pulposus cells were identified by using red O, toluidine blue, Swiss Giemsa staining and type II collagen immunohistochemical staining. After a period of starvation or LPS stimulation (4h, 16h, 24h, 48h), the expression of inflammatory related factors, IL-1 beta, IL-12, IL-17, IL-23, TNF- alpha, EBI3, was detected by RT-PCR method. The cultured cells were nucleus pulposus cells. Through PCR, the expression of IL-1 beta, IL-12, IL-17, IL-23, TNF- alpha and other inflammatory factors, such as EBI3 (IL-35) and IL-37, were also expressed in the basal state of the nucleus pulposus, and the expression of the nucleus was significantly higher than that in the basic state after starvation or LPS stimulation. 7, TNF- alpha was detected in cell culture, and also showed a similar expression. Conclusion: human medullary nucleus cells can not only express a variety of inflammatory factors, but also express anti-inflammatory factors. The expression of the cells is increased after starvation or LPS stimulation and is dependent on a certain time. Decomposition, angiogenesis, internal nerve growth, inflammatory progression, and pain sensitization play an important role. The understanding of the secretions and mechanisms of these inflammatory factors will help us to understand the biological characteristics of the intervertebral disc and provide new ideas and theoretical basis for the intervention or treatment of intervertebral disc related diseases.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R681.5

【參考文獻】

相關(guān)期刊論文 前5條

1 胡松;郭建極;劉濤;陳銘伍;冼磊;王永勇;周前;譚祥;;食管鱗癌患者血清IL-23和MMP-9的相關(guān)性及其臨床意義[J];實用醫(yī)學雜志;2014年18期

2 胡寶山;芮鋼;楊茂偉;呂剛;;白細胞介素-1β對椎間盤蛋白多糖代謝的影響[J];臨床和實驗醫(yī)學雜志;2008年01期

3 ;IL-1beta sensitizes rat intervertebral disc cells to Fas ligand mediated apoptosis in vitro[J];Acta Pharmacologica Sinica;2007年10期

4 王葵光，胡有谷;腰椎間盤突出癥的自身免疫狀態(tài)[J];中華骨科雜志;1994年05期

5 王濤;馬信龍;馬劍雄;田鵬;韓超;;兔腰椎間盤成分抗原性差異的研究[J];中華骨科雜志;2010年12期

相關(guān)碩士學位論文 前1條

1 田鵬;IL-17在腰椎間盤突出癥患者突出椎間盤組織中的表達[D];天津醫(yī)科大學;2010年

，

本文編號：1900536