發(fā)布時間：2018-05-20 15:38

  本文選題：椎間盤退變 + 鋅指蛋白A20��； 參考：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：目的:檢測鋅指蛋白A20、NF-κB在兔正常及退變腰椎間盤髓核組織中的表達,并在體外培養(yǎng)原代正常及退變的兔腰椎間盤髓核細胞,并進一步用內毒素(LPS)刺激髓核細胞,觀察鋅指蛋白A20、NF-κB及相關炎性因子的表達及其相互關系。方法:構建兔腰椎間盤退變動物模型,獲取兔腰椎間盤退變模型中正常及退變的椎間盤組織,HE染色觀察椎間盤髓核及纖維環(huán)的形態(tài)變化,免疫組化檢測椎間盤髓核細胞鋅指蛋白A20、NF-κB p65蛋白及Ⅱ型膠原(COL-Ⅱ)的表達。體外分離、培養(yǎng)獲取的正常和退變兔腰椎間盤髓核細胞,分為正常組、退變組、內毒素(LPS)刺激組和NF-κB抑制劑組,Real-time PCR分別檢測各組A20、IL-1β、TNF-α、NF-κB、Ⅱ型膠原mRNA的表達變化。Western blot觀察A20、p65、Ⅱ型膠原蛋白的表達變化。ELISA檢測細胞培養(yǎng)液中IL-1β、TNF-α的表達。結果:組織學觀察發(fā)現(xiàn)退變組髓核細胞數(shù)量明顯減少,髓核細胞聚集成團,Ⅱ型膠原表達下降,A20、p65蛋白表達升高;細胞實驗發(fā)現(xiàn),與正常組相比,退變組A20、TNF-α、IL-1β、p65表達在mRNA及蛋白水平均明顯升高,Ⅱ型膠原表達降低(p0.05);在LPS刺激組中A20、TNF-α、IL-1β、p65表達較退變組更為顯著,Ⅱ型膠原降低明顯(p0.05),而NF-κB抑制劑組A20、TNF-α、IL-1β、p65表達下降,Ⅱ型膠原表達回升,且與LPS刺激組有顯著統(tǒng)計學意義(p0.05)。結論:兔腰椎間盤退變時,鋅指蛋白A20表達增加,NF-κB活性增強;當加入LPS刺激后,鋅指蛋白A20、及其相關炎性因子表達進一步升高,且鋅指蛋白A20與NF-κB呈正相關,提示髓核細胞炎癥反應與兔椎間盤退變的發(fā)生、發(fā)展密切相關,其中A20可能發(fā)揮了重要作用。
[Abstract]:Objective: to detect the expression of zinc finger protein A20 (NF- 魏 B) in normal and degenerative nucleus pulposus of lumbar intervertebral disc in rabbits, and to culture primary normal and degenerative nucleus pulposus cells of rabbit lumbar intervertebral disc in vitro, and to further stimulate nucleus pulposus cells with endotoxin (LPS). To observe the expression and correlation of zinc finger protein A 20, NF- 魏 B and related inflammatory factors. Methods: a rabbit model of lumbar disc degeneration was established. The normal and degenerated intervertebral disc tissues were stained with HE to observe the morphological changes of the nucleus pulposus and the fibrous ring of the intervertebral disc. The expression of zinc finger protein A20, NF- 魏 B p65 and collagen 鈪，

本文編號：1915252