本文選題：家蠶 + Cu納米顆粒�。� 參考：《江蘇科技大學(xué)》2016年碩士論文

【摘要】：隨著納米材料的廣泛應(yīng)用,越來越多的納米產(chǎn)品進入到人們的生活中,極大的增加了其曝露于人體的機會。納米銅是一種常用的金屬納米材料,由于其特殊的物理化學(xué)特性,被廣泛的應(yīng)用在各領(lǐng)域。納米銅的廣泛應(yīng)用已引起了人們對其生物安全性的關(guān)注,目前關(guān)于納米銅毒性效應(yīng)的研究較少,其毒性機制尚不明確,同時納米銅的毒性試驗大都是以成本高、倫理道德爭議多的高級動物作為模型動物進行的,為了解決上述實驗中存在的問題并對納米銅的生物安全性評價提供理論依據(jù),尋找一種成本低、爭議少、對納米材料反應(yīng)靈敏的模型動物來研究納米銅的生物毒性顯得尤為重要。本研究以五齡家蠶作為模型動物,通過第二腹足注射曝露染毒,在200μg/頭劑量下研究納米銅對家蠶的毒性效應(yīng)和作用機制。實驗主要研究內(nèi)容和成果如下:1.分別給家蠶注射不同濃度的20 nm CuNPs懸浮液,通過曝露實驗結(jié)果計算得到CuNPs對家蠶的48 h半致死劑量(LD50)為196.05μg/頭,95%可置信區(qū)間為145.85?230.15μg/頭,CuNPs的曝露會抑制家蠶生長,影響家蠶食欲,同時也會給家蠶體內(nèi)組織帶來一定的損傷,并呈現(xiàn)一定的時間效應(yīng)和劑量效應(yīng)。2.建立了微波消解-電感耦合等離子體質(zhì)譜法檢測CuNPs在家蠶各組織含量的方法,實驗過程中Cu元素的標(biāo)準(zhǔn)曲線呈現(xiàn)良好的線性關(guān)系,空白組和實驗組中相關(guān)系數(shù)分別為0.99945和0.99969,Cu元素的加標(biāo)回收率在101%?106%之間,相對標(biāo)準(zhǔn)偏差RSD小于3.22%。實驗結(jié)果表明CuNPs在進入到家蠶體內(nèi)后會引起各組織中Cu元素的含量顯著性的升高(p0.05),其中脂肪體、中腸、氣管中Cu元素含量變化比較大,可能是CuNPs的主要靶器官。3.采用石蠟切片技術(shù)、TEM觀測及EDS能譜對家蠶各組織的病理變化和CuNPs的存在形態(tài)進行了分析。結(jié)果表明CuNPs在進入到家蠶體內(nèi)后會以原有的納米顆粒狀存在于家蠶各組織中,不但會破壞各組織自身的一些防御屏障對組織產(chǎn)生一些損傷,同時還會對家蠶各組織細胞產(chǎn)生一定的毒害。結(jié)果還發(fā)現(xiàn),進入家蠶組織內(nèi)的CuNPs會被一層生物分子所覆蓋,但對于這一覆蓋層的成分組成及相關(guān)的作用機理還不是很明確,需進一步的研究。4.在該曝露劑量下,CuNPs會對家蠶各組織產(chǎn)生一定的氧化損傷,表現(xiàn)為:家蠶各組織中ROS強度顯著提升(p0.05),同時氧化損傷標(biāo)志物脂質(zhì)過氧化產(chǎn)物MDA均顯著性提升(p0.05),DNA氧化產(chǎn)物8-OHdG水平也明顯升高;CuNPs還會破壞家蠶各組織的抗氧化系統(tǒng),表現(xiàn)為抗氧化酶SOD和CAT酶活性受到顯著性的抑制;各組織中環(huán)磷酸腺苷(cAMP)表達量顯著下降(p0.05)。5.在該曝露劑量下CuNPs會對家蠶各組織中細胞因子的表達產(chǎn)生一定的影響,破壞家蠶的免疫機制,并導(dǎo)致一系列的病變:各組織和血淋巴中半胱氨酸蛋白酶抑制劑(CysC)表達顯著性上調(diào)(p0.05)、核轉(zhuǎn)錄因子-κB(NF-κB)表達顯著性上調(diào)(p0.05)、免疫球蛋白(Ig)的分泌受到明顯抑制(p0.05)、細胞凋亡因子(Fas)顯著性上調(diào)(p0.05)。
[Abstract]:With the wide application of nanomaterials, more and more nanomaterials have entered people's lives and greatly increased their exposure to the human body. Nano copper is a kind of common metal nanomaterial. Because of its special physical and chemical properties, it has been widely used in the various domain. The wide application of nano copper has caused people to it. At present, there are few studies on the toxicity of nanoscale copper, and its toxicity mechanism is not clear. At the same time, the toxicity test of nanoscale copper is mostly based on high cost and ethical and moral disputes. In order to solve the problems in the experiment and evaluate the biosafety of nano copper, the toxicity of nano copper is not clear. It is very important to study the biological toxicity of nano copper by providing a theoretical basis, looking for a low cost and less controversial animal with sensitive reaction to nanomaterials to study the biological toxicity of nanoscale copper. In this study, five age silkworm was used as a model animal to study the toxicity and effect of nano copper on silkworm by injecting second gastropods and exposed to 200 mu g/ head. The main contents and results of the experiment are as follows: 1. the 20 nm CuNPs suspension of different concentration of silkworm was injected into the silkworm, and the 48 h semi lethal dose (LD50) of the silkworm (LD50) was 196.05 mu g/ head and 95% confidence interval was 145.85? 230.15 mu g/ head. The exposure of CuNPs to inhibit the growth of silkworm and affect the appetite of the silkworm. It also brings some damage to the tissues of silkworm, and presents a certain time effect and dose effect.2. to establish a method of microwave digestion inductively coupled plasma mass spectrometry to detect the content of CuNPs in the silkworm tissues. The standard curves of Cu elements in the experiment show a good linear relationship and the relationship between the blank group and the experimental group. The number of Cu elements was 0.99945 and 0.99969 respectively, and the recovery rate of the addition standard was 101%? 106%. The relative standard deviation RSD was less than 3.22%. experimental results showed that CuNPs could cause a significant increase in the content of Cu elements in the tissues of the silkworm (P0.05). The content of the Cu elements in the fat body, middle intestine and the trachea was relatively large, which may be CuNPs The main target organ of.3. was paraffin section technique, TEM observation and EDS spectrum analysis were used to analyze the pathological changes of the tissues of the silkworm and the existence form of CuNPs. The results showed that CuNPs would exist in the silkworm tissues in the silkworm body after entering the silkworm body, which not only destroyed some defense barrier pairs of the tissues themselves. It is also found that the CuNPs in the silkworm tissue will be covered by a layer of biomolecules, but the composition and the related mechanism of the covering layer are not very clear. It is necessary to study the.4. at the exposure dose of CuNPs. There were some oxidative damage in the tissues of the silkworm, which showed that the ROS intensity of the silkworm tissues increased significantly (P0.05), while the lipid peroxidation product MDA of the oxidative damage markers increased significantly (P0.05), and the 8-OHdG level of the DNA oxidation product increased obviously, and CuNPs also destroyed the antioxidant system of the silkworm tissues, which showed the antioxidant enzyme SOD and C. AT enzyme activity was significantly inhibited, and the expression of cAMP in each tissue decreased significantly (P0.05).5. at the exposed dose of CuNPs could affect the expression of cytokines in the tissues of the silkworm, destroying the immune mechanism of the silkworm, and causing a series of pathological changes: the inhibition of cysteine protease in the tissues and the hemolymph. The expression of CysC was significantly up-regulated (P0.05), the expression of nuclear factor kappa B (NF- kappa B) was significantly up-regulated (P0.05), the secretion of immunoglobulin (Ig) was significantly inhibited (P0.05), and apoptosis factor (Fas) was significantly up-regulated (P0.05).

【學(xué)位授予單位】：江蘇科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：X171.5;TB383.1

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