發(fā)布時間：2018-06-02 10:56

  本文選題：原子力顯微鏡 + 山奈酚��； 參考：《暨南大學(xué)》2012年碩士論文

【摘要】：免疫系統(tǒng)是人體對抗疾病，保護(hù)正常生理功能的重要系統(tǒng)之一，而疾病的發(fā)生、發(fā)展與免疫系統(tǒng)對其的反應(yīng)、變化息息相關(guān)。本文分別通過研究程序性凋亡配體1（PD-L1）及山奈酚對Jurkat T細(xì)胞的作用，闡釋細(xì)胞配體及藥物在免疫反應(yīng)中的作用，并得到以下結(jié)論： 1，，本文第二章研究了程序性凋亡配體PD-L1對于淋巴細(xì)胞的免疫抑制作用。通過CCK-8實驗指出程序性凋亡配體對于細(xì)胞增殖的抑制效果，該抑制效果隨濃度的增加而增強(qiáng)。通過Annexin V/PI細(xì)胞凋亡雙染試劑盒對細(xì)胞凋亡率進(jìn)行了檢測，發(fā)現(xiàn)細(xì)胞的凋亡率隨著配體濃度的增加而從4%增加到73%，進(jìn)一步佐證了CCK-8實驗的結(jié)論，指出凋亡配體對于細(xì)胞的誘導(dǎo)凋亡作用。實驗中還運(yùn)用流式細(xì)胞儀技術(shù)分析Jurkat T細(xì)胞表面早期活化分子CD69+和共刺激分子PD-1的表達(dá)變化情況，CD69+的表達(dá)從對照組細(xì)胞表面24%下降到實驗組細(xì)胞的1%，指出細(xì)胞的激活狀態(tài)被明顯抑制；而隨配體作用濃度的增加，PD-1的表達(dá)量也被誘導(dǎo)進(jìn)一步高表達(dá)，從對照組的1.2%增加到實驗組的7.3%。通過原子力顯微鏡對于對照組和實驗組的細(xì)胞進(jìn)行分析比較，發(fā)現(xiàn)凋亡配體作用后細(xì)胞的高度降低、超微結(jié)構(gòu)更加粗糙，進(jìn)一步解釋了細(xì)胞功能失活的現(xiàn)象。 2，本文第三章研究了山奈酚對于Jurkat T細(xì)胞的誘導(dǎo)凋亡作用。通過增殖抑制實驗發(fā)現(xiàn)山奈酚對于細(xì)胞增殖抑制效果成濃度與時間依賴性，我們通過細(xì)胞周期實驗發(fā)現(xiàn)實驗組中處于G2/M期的細(xì)胞所占比例隨著藥物作用濃度的增大也逐漸增大。我們進(jìn)一步對細(xì)胞內(nèi)Ca~(2+)的表達(dá)進(jìn)行了研究，發(fā)現(xiàn)鈣離子隨著藥物作用濃度的增加而從對照組的223(MFI)逐漸增加到實驗組的593(MFI)。鈣離子的大量釋放更加影響了細(xì)胞的活性，誘導(dǎo)其加速凋亡。之后通過分子對接實驗，模擬了Caspase3，F(xiàn)-actin分別與山奈酚作用的配位情況，指出其潛在的作用位點。最后，通過原子力顯微鏡對于對照組和實驗組的細(xì)胞進(jìn)行分析比較，指出山奈酚作用后細(xì)胞的胞體遭到破壞，力學(xué)性質(zhì)等信息產(chǎn)生了明顯變化，粘附力和楊氏模量均變小，以上的數(shù)據(jù)共同解釋了細(xì)胞功能失活的現(xiàn)象。
[Abstract]:The immune system is one of the important systems for human body to fight diseases and protect normal physiological functions. The occurrence and development of diseases are closely related to the response and changes of the immune system to them. In order to elucidate the role of cell ligands and drugs in immune response, we studied the effects of procedural apoptotic ligand 1 (PD-L 1) and kaempferol on Jurkat T cells, and obtained the following conclusions: 1. In chapter 2, the immunosuppressive effect of programmed apoptotic ligand PD-L1 on lymphocytes was studied. The inhibitory effect of programmed apoptotic ligand on cell proliferation was indicated by CCK-8 assay, and the inhibitory effect increased with the increase of concentration. The apoptosis rate of Annexin V/PI cells was detected by double staining kit. It was found that the apoptosis rate increased from 4% to 73% with the increase of ligand concentration, which further confirmed the conclusion of CCK-8 experiment. The effect of apoptotic ligand on cell apoptosis was pointed out. Flow cytometry was also used to analyze the expression of early activated molecule CD69 and costimulatory molecule PD-1 on the surface of Jurkat T cells. The expression of CD69 decreased from 24% on the surface of the control group to 1 cell in the experimental group. The activation state was obviously inhibited. With the increase of ligand concentration, the expression of PD-1 was further increased from 1.2% in the control group to 7.3% in the experimental group. The results of atomic force microscope (AFM) showed that the cell height decreased and the ultrastructure became rougher after the action of apoptotic ligand, which further explained the phenomenon of cell inactivation. 2. In chapter 3, we studied the apoptosis of Jurkat T cells induced by kaempferol. The inhibitory effect of kaempferol on cell proliferation was found to be concentration-dependent and time-dependent. We found that the proportion of cells in G _ 2 / M phase in experimental group increased with the increase of drug concentration. We further studied the expression of Ca~(2 in cells. It was found that with the increase of the concentration of calcium, calcium gradually increased from 223m MFI in the control group to 593 渭 m MFI in the experimental group. The release of Ca ~ (2 +) significantly affected cell activity and induced apoptosis. The coordination of Caspase3Factin with kaempferol was simulated by molecular docking experiments, and the potential sites of Caspase3 F-actin were pointed out. Finally, the cells of the control group and experimental group were analyzed and compared by atomic force microscope. It was pointed out that the cell body was destroyed, the mechanical properties of the cells were obviously changed, and the adhesion force and Young's modulus were both decreased after the treatment of kaempferol. The above data together explain the inactivation of cell function.
【學(xué)位授予單位】：暨南大學(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)碩士學(xué)位論文 前1條

1 慕靜靜;山奈酚對小鼠免疫功能的影響及對海仁酸致癇小鼠的免疫調(diào)節(jié)作用[D];暨南大學(xué);2010年

本文編號：1968508