本文關(guān)鍵詞： 黑蒜 Lewis Ht-29 放射增敏 分子機制　出處：《濱州醫(yī)學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：觀察黑蒜提取液(Aged Black Garlic Extract-ABGE)對Lewis市癌細胞株及結(jié)腸癌Ht-29小鼠移植瘤的放射增敏作用并探討其可能機制。方法：常規(guī)培養(yǎng)lewis肺癌細胞,用不同濃度的黑蒜提取液聯(lián)合6oCoγ線照射進行體外實驗。MTT法測定黑蒜提取液對肺癌lewis細胞株生長抑制率；平板集落形成試驗觀察黑蒜提取液聯(lián)合放療對細胞克隆存活的影響；Hoechst熒光染色觀察細胞形態(tài)的變化；用流式細胞儀檢測黑蒜提取液作用后腫瘤細胞周期變化；Western Blot分析黑蒜提取液對lewis細胞Bcl-2、Bax蛋白表達影響。建立結(jié)腸癌Ht-29小鼠皮下移植瘤模型,將40只荷瘤小鼠隨機分為4組,空白對照組,γ照射組,黑蒜組,黑蒜+γ照射組。分別給予生理鹽水、γ射線照射、黑蒜提取液及黑蒜提取液聯(lián)合γ射線照射的處理,觀察各組移植瘤生長速度和各治療組移植瘤生長延遲時間。實驗結(jié)束后摘取眼球取血后脫臼處死小鼠,剝離瘤體稱重,計算腫瘤生長抑制率；計算腫瘤生長延遲時間及放射增敏比；流式細胞術(shù)Annexin V-FITC/PI雙染法檢測腫瘤組織細胞凋亡情況；Realtime-PCR檢測Bcl-2、Bax、Caspase-9的表達情況；外周血白細胞計數(shù)、紫外分光光度計法檢測各組小鼠血清中SOD及CAT的活力變化。結(jié)果：(1)黑蒜提取液對肺癌lewis細胞株生長有明顯抑制作用,黑蒜提取液聯(lián)合60 Co γ射線照射能夠促進lewis細胞凋亡,影響細胞周期及Bcl-2、Bax蛋白表達。(2)黑蒜提取液聯(lián)合γ線照射對結(jié)腸癌Ht-29小鼠移植瘤有生長抑制作用,可延遲腫瘤生長時間,提高放療增敏比,促進腫瘤細胞凋亡,并且能夠下調(diào)移植瘤組織中Bcl-2,上調(diào)Bax,提高Caspase-9蛋白表達。黑蒜提取液可提高小鼠的外周血白細胞總數(shù),增加紅細胞超氧化物歧化酶SOD及CAT的活力。結(jié)論：(1)黑蒜提取液對肺癌lewis細胞具有放射增敏作用,其增敏機制可能與其改變細胞周期及影響B(tài) cl-2、B ax蛋白表達有關(guān)。(2)黑蒜提取液可通過下調(diào)Bcl-2上調(diào)Bax及活化Caspase-9蛋白的表達提高γ射線對結(jié)腸癌Ht-29小鼠移植瘤放射敏感性,并通過提高外周血白細胞計數(shù)和機體抗氧化能力提高對機體的輻射保護作用。
[Abstract]:Objective: to observe the radiosensitizing effect of Aged Black Garlic extract (Aged Black Garlic Extract-ABGE) on Lewis cancer cell line and colonic cancer Ht-29 mouse transplanted tumor and to explore its possible mechanism. Methods: lewis lung cancer cells were cultured routinely. Different concentrations of black garlic extract combined with 6o Co 緯 -ray irradiation were used to determine the growth inhibition rate of black garlic extract on lung cancer lewis cell line in vitro. The effect of black garlic extract combined with radiotherapy on cell clone survival was observed by plate colony forming test. The morphological changes of cells were observed by Hoechst fluorescence staining. Flow cytometry was used to detect the changes of tumor cell cycle after the treatment of black garlic extract. Western Blot was used to analyze the effect of black garlic extract on the expression of Bcl-2P Bax protein in lewis cells. A subcutaneous transplanted tumor model of Ht-29 mice with colon cancer was established and 40 mice were randomly divided into 4 groups. Blank control group, 緯 irradiation group, black garlic group and black garlic 緯 irradiation group were treated with normal saline, 緯 -ray irradiation, black garlic extract and black garlic extract combined with 緯 -ray irradiation, respectively. The growth rate of transplanted tumor in each group and the growth delay time of each treatment group were observed. After the experiment, the mice were killed after eyeball extraction, the tumor was weighed and the tumor growth inhibition rate was calculated. The tumor growth delay time and radiosensitization ratio were calculated, the apoptosis of tumor tissue was detected by flow cytometry with Annexin V-FITC / Pi double staining, and the expression of Bcl-2BaxCaspase-9 was detected by Realtime-PCR. The activity of SOD and CAT in serum of each group of mice was detected by ultraviolet spectrophotometer. Results the extract of black garlic significantly inhibited the growth of lung cancer lewis cell line, and the combination of black garlic extract and 60Co 緯 -ray irradiation could promote the apoptosis of lewis cells. The cell cycle and the expression of Bcl-2P Bax protein.) Black garlic extract combined with 緯 -ray irradiation can inhibit the growth of transplanted tumor of colon cancer Ht-29 mice, delay the tumor growth time, increase the radiosensitization ratio and promote the apoptosis of tumor cells. It can down-regulate Bcl-2, up-regulate Baxand increase the expression of Caspase-9 protein. The extract of black garlic can increase the total number of white blood cells in peripheral blood of mice. Conclusion the extract of RBC superoxide dismutase (SOD) and CAT can enhance the radiosensitivity of lung cancer lewis cells. The mechanism of its sensitizing mechanism may be related to its alteration of cell cycle and its effect on the expression of Bcl-2 Bax protein. The extract of black garlic can increase the radiosensitivity to colonic cancer Ht-29 transplanted tumor by down-regulating the expression of Bax and activating Caspase-9 protein by down-regulating Bcl-2, and increasing the radiosensitivity of 緯 -ray to colonic cancer Ht-29 mice. The radiation protection of the body was improved by increasing the white blood cell count and anti-oxidation ability of the body.
【學(xué)位授予單位】：濱州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R730.55

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本文編號：1502808