發(fā)布時間：2018-05-14 15:20

  本文選題：乳鐵蛋白 + 輻射損傷�。� 參考：《濟南大學》2013年碩士論文

【摘要】：目的： 隨著核技術(shù)的迅速發(fā)展，電離輻射的應(yīng)用已經(jīng)發(fā)展到各個領(lǐng)域。工業(yè)、科研和醫(yī)學等領(lǐng)域使用的X射線探傷機、放射性同位素、X射線機、直線加速器等，一旦造成事故就會給人體造成不同程度的損傷。人體的多個系統(tǒng)如造血系統(tǒng)、免疫系統(tǒng)和生殖系統(tǒng)對電離輻射非常敏感。因此，人體長期或者大劑量接受電離輻射，會引起細胞、組織和器官損傷，誘發(fā)各種疾病，，誘發(fā)癌癥、白血病，甚至引起死亡。乳鐵蛋白是一種非血紅素鐵結(jié)合性糖蛋白，廣泛分布于哺乳動物的乳汁、唾液、淚液等分泌物和血液、嗜中性粒細胞中。乳鐵蛋白已在多個領(lǐng)域表現(xiàn)出特有的生物活性，如促進腸道鐵吸收、抑菌與殺菌、抗氧化作用、免疫調(diào)節(jié)作用、促進腸道內(nèi)雙歧桿菌生長功能、抗感染作用、抗癌作用、藥物協(xié)同作用。2006年，日本研究人員通過動物實驗證實，乳鐵蛋白具有防止輻射損傷的功效，并認為這種作用與乳鐵蛋白的抗氧化作用有關(guān)。本課題通過研究乳鐵蛋白對輻射損傷小鼠抗氧化、免疫功能、細胞凋亡等的影響，進一步探討乳鐵蛋白對小鼠的抗輻射損傷作用及可能存在的機制，為研究新型放射防護劑提供新的思路和方法。 方法： 首先研究乳鐵蛋白對小鼠輻射損傷的防護作用。36只SPF級Balb/c小鼠適應(yīng)性飼養(yǎng)1周，觀察無異常后參照隨機區(qū)組法分組，隨機分為正常對照組、照射對照組和hLF組，每組12只，照射對照組和hLF組給予2Gy X射線全身照射。照射后hLF組每只小鼠腹腔注射4.0mg/ml的乳鐵蛋白溶液1.0ml；正常對照組和照射對照組腹腔注射0.9％氯化鈉注射液各1.0ml，各組小鼠自由飲食。照射后24h，處死小鼠取材，檢測免疫器官指數(shù)、T淋巴細胞亞群比例、血清SOD活力、MDA水平、外周血象。另取Balb/c小鼠30只，分組、照射、給藥及飲食方式同上述實驗。給藥20h后，每組小鼠腹腔注射體積分數(shù)為5％雞紅細胞懸液1.0ml，4h后，取材，檢測腹腔巨噬細胞吞噬功能。 第二是研究乳鐵蛋白對脾細胞凋亡和細胞因子的影響。取Balb/c小鼠30只，分組同上述實驗。hLF組按50mg/kg hLF灌胃給藥，1次/d，連續(xù)7d；正常對照組和照射對照組灌胃0.9％氯化鈉注射液，20ml/kg，1次/d，共7d。末次給藥后2h，照射對照組和hLF組給予2Gy X射線全身照射，各組小鼠自由飲食。照射后24h，處死小鼠取材，檢測脾細胞凋亡、血清Bcl-2、IL-1β、IL-2。 結(jié)果： 乳鐵蛋白對小鼠輻射損傷的防護作用實驗。實驗結(jié)果發(fā)現(xiàn)，照射后，照射對照組和hLF組小鼠與正常對照組比較，小鼠胸腺和脾臟指數(shù)、CD8+淋巴細胞比例、SOD活力、WBC、LYM％、腹腔巨噬細胞吞噬指數(shù)和吞噬百分率均顯著降低（P0.01），而MDA水平、CD4+淋巴細胞比例均顯著增高，差異有統(tǒng)計學意義（P0.01）；hLF組小鼠脾臟和胸腺指數(shù)、CD4+、CD8+淋巴細胞比例、SOD活力、LYM％、腹腔巨噬細胞吞噬指數(shù)和吞噬百分率明顯高于照射對照組（P0.01，P0.05），而MDA水平顯著低于照射對照組，差異有統(tǒng)計學意義（P0.01）。 乳鐵蛋白對脾細胞凋亡和細胞因子的影響實驗。實驗結(jié)果發(fā)現(xiàn)，照射后，小鼠脾細胞凋亡增多，照射對照組小鼠脾細胞凋亡的細胞比例（包括早期凋亡細胞和晚期凋亡細胞）顯著高于正常對照組，差異具有統(tǒng)計學意義（P0.05）；hLF組小鼠脾細胞凋亡細胞的比例正常對照組有所升高，差異無統(tǒng)計學意義（P0.05）；hLF組小鼠脾細胞凋亡細胞的比例顯著低于照射對照組，差異有統(tǒng)計學意義（P0.01）。hLF組小鼠血清Bcl-2、IL-1β、IL-2濃度顯著高于照射對照組，差異有統(tǒng)計學意義(P0.05)。 結(jié)論： 1.免疫器官指數(shù)計算結(jié)果提示乳鐵蛋白可能能夠防止電離輻射引起的胸腺和脾臟的過度損傷并促進其恢復。 2.脾臟T淋巴細胞亞群和腹腔巨噬細胞吞噬實驗證實乳鐵蛋白對電離輻射引起的免疫功能損傷具有保護作用。 3.抗氧化實驗的結(jié)果提示乳鐵蛋白可提高電離輻射損傷后小鼠機體內(nèi)清除自由基的能力和抗氧化能力。 4.乳鐵蛋白提高輻射損傷小鼠外周血LYM％，但是對WBC、RBC、PLT等無明顯的保護作用，這還需要進一步的研究。 5.脾淋巴細胞凋亡和血清Bcl-2測定結(jié)果表明乳鐵蛋白可能通過提高bcl-2的表達，發(fā)揮抑制細胞凋亡的作用。 6.IL-1β、IL-2測定結(jié)果表明乳鐵蛋白能夠增強IL-1β、IL-2的分泌來抑制輻射后機體的免疫功能的損傷。
[Abstract]:Purpose :

With the rapid development of nuclear technology , the application of ionizing radiation has been developed into various fields , such as X - ray detectors , radioactive isotopes , X - ray machines and linear accelerators used in the fields of industrial , scientific research and medicine .

Method :

The protective effects of lactoferrin on radiation injury in mice were studied . Thirty - six SPF - grade Balb / c mice were randomly divided into two groups : normal control group , control group and hLF group .
In the normal control group and the control group , 0.9 % sodium chloride injection was injected intraperitoneally with 1.0 ml each group . The mice were sacrificed 24 hours after irradiation . The mice were sacrificed for 24 h , the proportion of T lymphocyte subsets , serum SOD activity , MDA level and peripheral blood image were detected .

The second was to study the effect of lactoferrin on apoptosis and cytokines in spleen cells . Balb / c mice were divided into 30 groups and group with the above - mentioned experiment . hLF group was given orally at 50mg / kg hLF , and lasted for 7 days .
In the normal control group and the control group , 0.9 % sodium chloride injection , 20 ml / kg , 1 time / day were injected into the control group for 7 days . 2 hours after the last dose , 2 Gy X - ray whole body irradiation was given to the control group and the hLF group . After irradiation , the mice were irradiated for 24 hours , the mice were sacrificed and the apoptosis of the spleen cells , the serum Bcl - 2 , IL - 1尾 and IL - 2 were detected .

Results :

The protective effects of lactoferrin on radiation injury in mice were observed . The results showed that the percentage of thymus and spleen index , percentage of CD8 + lymphocytes , SOD activity , WBC , LYM % , phagocytic index and phagocytic percentage of peritoneal macrophages decreased significantly after irradiation ( P0.01 ) , while the levels of MDA and CD4 + lymphocytes increased significantly ( P0.01 ) .
In hLF group , spleen and thymus index , CD4 + , CD8 + lymphocyte ratio , SOD activity , LYM % , phagocytic index and phagocytic percentage of peritoneal macrophages were significantly higher than those in control group ( P0.01 , P0.05 ) , while MDA level was significantly lower than that in control group ( P0.01 ) .

The effects of lactoferrin on apoptosis and cytokines in spleen cells showed that the apoptosis of spleen cells increased after irradiation , and the percentage of apoptosis in spleen cells ( including early apoptotic cells and late apoptotic cells ) was significantly higher in the irradiated mice than in the normal control group ( P0.05 ) .
The percentage of apoptotic cells in spleen cells of hLF group was higher than that of control group ( P0.05 ) .
The percentage of apoptotic cells in spleen cells of hLF group was significantly lower than that in control group ( P0.01 ) . The levels of Bcl - 2 , IL - 1尾 and IL - 2 in serum of hLF group were significantly higher than those in control group ( P0.05 ) .

Conclusion :

1 . The results of the immune organ index suggest that lactoferrin may be capable of preventing excessive damage of thymus and spleen caused by ionizing radiation and promoting its recovery .

2 . The spleen T lymphocyte subsets and the phagocytosis of peritoneal macrophages confirmed that lactoferrin had a protective effect on the immune function injury induced by ionizing radiation .

3 . The results of anti - oxidation experiments suggested that lactoferrin could enhance the ability of scavenging free radicals and anti - oxidation ability in mice after ionizing radiation injury .

4 . Milk ferritin increased lyM % in peripheral blood of irradiated mice , but there was no obvious protective effect on WBC , RBC , PLT .

5 . Apoptosis of splenic lymphocytes and the determination of Bcl - 2 in serum showed that lactoferrin could play a role in inhibiting apoptosis by increasing the expression of bcl - 2 .

6 . IL - 1尾 , IL - 2 assay showed that lactoferrin could enhance the secretion of IL - 1尾 and IL - 2 to inhibit the immune function of IL - 1尾 and IL - 2 .

【學位授予單位】：濟南大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R142

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