本文選題：迷迭香酸 + NF-κB��； 參考：《南華大學(xué)》2012年碩士論文

【摘要】：目的： 觀察迷迭香酸（RA）對抗谷氨酸誘導(dǎo)PC12細(xì)胞凋亡發(fā)揮神經(jīng)保護(hù)作用，探討作用機(jī)制與核轉(zhuǎn)錄因子-κB（NF-κB）信號通路的關(guān)系。 方法： 體外培養(yǎng)PC12細(xì)胞，建立谷氨酸誘導(dǎo)細(xì)胞凋亡模型。采用MTT法檢測細(xì)胞存活率；Hoechst33258熒光染色觀察細(xì)胞凋亡形態(tài)的改變，碘化丙啶（PI）染色流式細(xì)胞儀檢測分析細(xì)胞凋亡率；Western blot檢測IκBα、p-IκBα蛋白、NF-κBp65蛋白在細(xì)胞漿和細(xì)胞核中表達(dá)變化情況。 結(jié)果： 不同濃度谷氨酸（0、4、8、12、16、20mmol/L）作用PC12細(xì)胞24h后，細(xì)胞存活率呈劑量依賴性下降，其IC75大約為16mmol/L。因此，本實(shí)驗(yàn)以16mmol/L谷氨酸作為PC12細(xì)胞損傷模型的工作濃度，，將后續(xù)實(shí)驗(yàn)分組為：對照組，損傷組（16mmol/L谷氨酸），RA1組（30μmol/L RA+16mmol/L谷氨酸），RA2組（60μmol/L RA+16mmol/L谷氨酸）。16mmol/L谷氨酸孵育PC12細(xì)胞24h后，細(xì)胞核形態(tài)出現(xiàn)凋亡小體、細(xì)胞核濃縮或碎塊狀等典型凋亡樣改變，細(xì)胞凋亡率為(28.6±2.5)%；與損傷組比較，RA1組和RA2組細(xì)胞存活率隨RA濃度增加明顯升高（p 0.05），并能減少細(xì)胞凋亡發(fā)生，使其凋亡率分別下降至(18.3±3.7)%、(7.8±1.9)%。16mmol/L谷氨酸作用PC12細(xì)胞1h后，細(xì)胞漿中IκBα蛋白和NF-κB p65蛋白減少，p-IκBα蛋白表達(dá)增加，NF-κB p65蛋白在細(xì)胞核中表達(dá)增加，2h達(dá)到高峰；給予30、60μmol/L RA預(yù)處理1h后，能抑制NF-κBp65蛋白活化進(jìn)入細(xì)胞核。 結(jié)論： 1、在PC12細(xì)胞模型中，迷迭香酸有抗谷氨酸誘導(dǎo)細(xì)胞凋亡的作用； 2、 RA對抗谷氨酸毒性發(fā)揮神經(jīng)保護(hù)作用的機(jī)制，可能與抑制NF-κB活性有關(guān)。
[Abstract]:Objective: To observe the neuroprotective effect of rosmarinic acid (RAA) on glutamate induced apoptosis of PC12 cells, and to explore the relationship between the mechanism and the signal pathway of nuclear transcription factor-魏 B (NF- 魏 B) NF- 魏 B. Methods: PC12 cells were cultured in vitro, and glutamate induced apoptosis model was established. The morphology of cell apoptosis was observed by MTT assay. The apoptosis rate was detected by flow cytometry. The expression of NF- 魏 Bp65 in cytoplasm and nucleus of I 魏 B 偽 -p-I 魏 B 偽 protein was detected by Western blot. Results: The survival rate of PC12 cells decreased in a dose-dependent manner after 24 h treatment with different concentrations of Glutamic acid. The IC75 was about 16 mmol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1). Therefore, in this experiment, 16mmol/L glutamate was used as the working concentration of PC12 cell injury model. The following experiments were divided into two groups: control group. The injured group was divided into two groups: the injured group was treated with 60 渭 mol/L RA 16mmol/L glutamate, 16 mmol / L glutamate / L glutamic acid for 24 hours, and the injured group was treated with 60 渭 mol/L RA 16mmol/L glutamate, 16 mmol / L glutamic acid for 24 hours after incubating PC12 cells with 60 渭 mol/L RA 16mmol/L glutamate or 16 mmol / L glutamate. The apoptotic bodies appeared in nuclear morphology, the apoptotic rate was 28.6 鹵2.5g, the cell survival rate of RA1 group and RA2 group increased significantly with the increase of RA concentration, and the apoptosis rate decreased with the increase of RA concentration. After treated with glutamate of 7.8 鹵1.9)%.16mmol/L for 1 h, I 魏 B 偽 protein and NF- 魏 B p65 protein in the cytoplasm decreased the expression of p-I 魏 B 偽 protein and increased the expression of NF- 魏 B p65 protein in the nucleus of PC12 cells. The expression of NF- 魏 B p65 protein increased to a peak at 2 h after pretreatment with 3060 渭 mol/L RA for 1 h. It can inhibit the activation of NF- 魏 Bp65 protein into the nucleus. Conclusion: 1. In PC12 cell model, rosemary acid could inhibit the apoptosis induced by glutamate. 2. The neuroprotective mechanism of RA against glutamate toxicity may be related to the inhibition of NF- 魏 B activity.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 黃幼霞;黃榮桂;鄭興中;;迷迭香酸藥理作用的研究進(jìn)展[J];海峽藥學(xué);2010年05期

2 ;Dual roles of NF-kB in cell survival and implications of NF-kB inhibitors in neuroprotective therapy[J];Acta Pharmacologica Sinica;2007年12期

3 焦俊霞;高維娟;;細(xì)胞凋亡的信號轉(zhuǎn)導(dǎo)機(jī)制研究進(jìn)展[J];中國老年學(xué)雜志;2010年06期

本文編號：1808280