本文選題：椎間盤退變模型 + 體外培養(yǎng)　； 參考：《南京醫(yī)科大學(xué)》2015年碩士論文

【摘要】：[目的]:建立小鼠椎間盤體外培養(yǎng)退變模型,分析抗氧化劑N-乙�；�-L-半胱氨酸(NAC)對(duì)椎間盤退變的影響。[方法]:選取8周齡ICR小鼠作為實(shí)驗(yàn)對(duì)象,無(wú)菌條件下取出小鼠腰段椎間盤,隨機(jī)分為三組:對(duì)照組、退變模型組及NAC處理組,在退變模型組中的培養(yǎng)基加入白細(xì)胞介素(IL)1β和腫瘤壞死因子(TNF)α,NAC處理組的培養(yǎng)基中除了加入上述炎癥因子外,再加入配制好的NAC溶液。在培養(yǎng)液中經(jīng)過(guò)不同時(shí)段培養(yǎng)后,分別于3天、7天、10天取出培養(yǎng)液中的椎間盤。通過(guò)組織學(xué)和免疫組織化學(xué)等方法分析椎間盤形態(tài)學(xué)特征,并提取椎間盤組織中的蛋白和RNA,分別通過(guò)Western-blot和RT-PCR方法檢測(cè)椎間盤退變過(guò)程中的相關(guān)蛋白表達(dá)和基因轉(zhuǎn)錄水平。[結(jié)果]:與退變模型組相比,培養(yǎng)3、7、10天的NAC處理組椎間盤經(jīng)HE-阿爾新藍(lán)復(fù)合染色顯示其結(jié)構(gòu)逐漸分界清楚,蛋白多糖含量豐富;二型膠原(Col-Ⅱ)、十型膠原(Col-X)及基質(zhì)金屬蛋白酶-3(MMP-3)表達(dá)均降低;抗氧化蛋白SOD-1表達(dá)升高,抗氧化相關(guān)基因GSR、Gpx4表達(dá)水平升高;增殖相關(guān)蛋白Cyclin D1表達(dá)升高;促凋亡蛋白Caspase-3表達(dá)降低,促凋亡相關(guān)基因nox A、Perp表達(dá)水平降低;NAC處理組上述各項(xiàng)指標(biāo)均接近對(duì)照組水平。[結(jié)論]:通過(guò)在培養(yǎng)基中加入炎癥因子IL-1β和TNF-α,可以成功建立椎間盤體外培養(yǎng)退變模型�？寡趸瘎㎞AC補(bǔ)充可以通過(guò)降低氧化應(yīng)激,促進(jìn)細(xì)胞增殖,減少細(xì)胞凋亡延緩椎間盤退變。
[Abstract]:[Objective] to establish an in vitro culture degenerative model of the mouse intervertebral disc and analyze the effect of antioxidant N- acetyl -L- cysteine (NAC) on the degeneration of intervertebral disc. [Methods] the 8 week old ICR mice were selected as the experimental subjects, and the lumbar intervertebral discs were taken out under aseptic conditions and divided into three groups randomly: the control group, the degeneration model group and the NAC treatment group, in the degenerative model. The medium in the group was added to interleukin (IL) 1 beta and tumor necrosis factor (TNF) alpha. The medium of NAC treatment group was added to the prepared NAC solution in addition to the above inflammatory factors. After different periods of culture in the culture medium, the intervertebral discs were removed from the culture solution at 3 days, 7 days and 10 days respectively. Chemical and other methods were used to analyze the morphological characteristics of intervertebral disc, and extract the protein and RNA in the intervertebral disc tissue. The protein expression and gene transcription level in the process of intervertebral disc degeneration were detected by Western-blot and RT-PCR. [results]: compared with the degenerative model group, the NAC treatment group of NAC treatment group with 3,7,10 days was mixed with alnew blue. The staining showed that the structure gradually became clear and the content of proteoglycan was rich; the expression of type two collagen (Col- II), type ten collagen (Col-X) and matrix metalloproteinase -3 (MMP-3) decreased, the expression of antioxidant protein SOD-1 increased, the expression level of antioxidant related gene GSR, Gpx4 increased, the D1 expression of proliferation related protein Cyclin increased, and apoptotic protein Caspase- was increased. The expression of 3 was reduced, the expression of apoptosis related genes NOx A and Perp decreased, and the above indexes in the NAC treatment group were close to the control level. [Conclusion]: by adding the inflammatory factor IL-1 beta and TNF- alpha in the medium, the model of the intervertebral disc in vitro culture degenerative model can be successfully established. The antioxidant NAC supplement can promote the cell by reducing oxidative stress. Proliferation, reducing apoptosis and retarding intervertebral disc degeneration.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R681.53

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