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

【摘要】：目的： 觀察迷迭香酸（RA）對抗谷氨酸誘導PC12細胞凋亡發(fā)揮神經保護作用，探討作用機制與核轉錄因子-κB（NF-κB）信號通路的關系。 方法： 體外培養(yǎng)PC12細胞，建立谷氨酸誘導細胞凋亡模型。采用MTT法檢測細胞存活率；Hoechst33258熒光染色觀察細胞凋亡形態(tài)的改變，碘化丙啶（PI）染色流式細胞儀檢測分析細胞凋亡率；Western blot檢測IκBα、p-IκBα蛋白、NF-κBp65蛋白在細胞漿和細胞核中表達變化情況。 結果： 不同濃度谷氨酸（0、4、8、12、16、20mmol/L）作用PC12細胞24h后，細胞存活率呈劑量依賴性下降，其IC75大約為16mmol/L。因此，本實驗以16mmol/L谷氨酸作為PC12細胞損傷模型的工作濃度，，將后續(xù)實驗分組為：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細胞24h后，細胞核形態(tài)出現(xiàn)凋亡小體、細胞核濃縮或碎塊狀等典型凋亡樣改變，細胞凋亡率為(28.6±2.5)%；與損傷組比較，RA1組和RA2組細胞存活率隨RA濃度增加明顯升高（p 0.05），并能減少細胞凋亡發(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細胞1h后，細胞漿中IκBα蛋白和NF-κB p65蛋白減少，p-IκBα蛋白表達增加，NF-κB p65蛋白在細胞核中表達增加，2h達到高峰；給予30、60μmol/L RA預處理1h后，能抑制NF-κBp65蛋白活化進入細胞核。 結論： 1、在PC12細胞模型中，迷迭香酸有抗谷氨酸誘導細胞凋亡的作用； 2、 RA對抗谷氨酸毒性發(fā)揮神經保護作用的機制，可能與抑制NF-κB活性有關。
[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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363

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相關期刊論文 前3條

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3 焦俊霞;高維娟;;細胞凋亡的信號轉導機制研究進展[J];中國老年學雜志;2010年06期

本文編號：1808280