本文選題：大鼠 + 原發(fā)性肝癌��； 參考：《揚州大學(xué)》2014年碩士論文

【摘要】：在我國肝細(xì)胞癌(Hepatocellular, HCC)的發(fā)病率居全球首位,其死亡率居惡性腫瘤死亡的第二位。肝癌的早期發(fā)現(xiàn)、早期診斷決定著其臨床治療的效果及患者預(yù)后。臨床常用的檢測手段包括血清AFP檢測和各種影像學(xué)手段。甲胎蛋白(Alpha-fetoprotein, AFP)是HCC特異性標(biāo)志物,80%的患者可出現(xiàn)甲胎蛋白的升高。如果將抗AFP抗體與某些顯影劑或(和)抗癌藥物相連,可以實現(xiàn)對肝癌的靶向成像和靶向治療,使得影像診斷和臨床治療的同時進行成為可能。本實驗擬通過對小劑量二乙基亞硝胺(DENA)誘導(dǎo)SD大鼠原位肝癌模型的MR分子成像、動態(tài)觀察和病理學(xué)對照,研究靶向AFP的超小型超順磁性納米粒子在HCC中的特異性靶向成像和分布規(guī)律,本研究主要包括兩部分。 第一部分：sD大鼠肝癌模型的建立及靶向AFP-USPIO在SD大鼠肝癌中的MR成像表現(xiàn) 目的：建立SD大鼠肝癌模型,進行AFP抗體標(biāo)記USPIO增強掃描,觀察大鼠肝癌特異性MRI成像。 方法：定期配制濃度為0.1mg/ml的DENA水溶液,予25只雄性SD大鼠自由飲用,喂養(yǎng)14周后改為空白飲用水。16-18周時選取利于實驗觀察的大鼠肝癌模型20只,完全隨機分成2組：實驗組10只,對照組10只。所有大鼠先行MRI T2WI平掃,實驗組注入AFP-USPIO(靶向組),對照組注入USPIO(非靶向組)2h后行T2WI增強掃描,觀察病灶的信號變化情況,測量增強前后病灶、肝臟的信號強度,計算其對比噪聲比(CNR)；處死大鼠后,取病灶標(biāo)本行HE、AFP免疫組化染色和普魯士藍染色分析。 結(jié)果：MRI掃描T2WI示肝內(nèi)多發(fā)大小不等的高、稍高信號及混雜信號結(jié)節(jié)影,選取直徑3mmm的結(jié)節(jié)作為實驗對象進行觀察。病灶境界顯示清楚,大部分信號均勻,呈高、稍高信號。實驗組、對照組分別注入AFP-USPIO、USPIO后2h增強掃描,在注射AFP-USPIO前后,大鼠肝臟-腫瘤的CNR分別為10.0+2.45和4.73士2.51,差異有統(tǒng)計學(xué)意義(P0.001,t=11.23)：而注射USPIO前后肝臟-腫瘤的CNR分別為9.15+1.24和9.96±1.63,差異無統(tǒng)計學(xué)意義(P=0.186,t=-1.43)。 病理結(jié)果HE染色顯示,選取作為MRI觀察的病灶均為肝細(xì)胞肝癌, AFP免疫組織化學(xué)結(jié)果顯示肝癌細(xì)胞胞漿內(nèi)大量表達AFP。普魯士藍鐵染色結(jié)果顯示,實驗組腫瘤組織間隙及腫瘤細(xì)胞內(nèi)見較多藍染顆粒,對照組腫瘤組織內(nèi)藍染顆粒則較少。 結(jié)論：二乙基亞硝胺誘發(fā)SD大鼠肝癌的模型建立方便,成瘤率高。增強后AFP-USPIO靶向造影劑在大鼠肝癌組織聚集,降低腫瘤組織的信號,雖然腫瘤肝臟的對比噪聲比減低,但有助于肝癌的定性診斷。 第二部分：AFP-USPIO在大鼠肝癌組織中的分布變化的實驗研究 目的：研究抗體標(biāo)記超順磁性氧化鐵納米粒(AFP-USPIO)在大鼠肝癌組織中隨時間的分布變化情況,為下一步偶聯(lián)化療藥物的納米粒子在腫瘤內(nèi)的分布提供理論支持。 方法：建立SD大鼠肝癌模型10只,方法及要求同上。先進行MR T2WI平掃,再分別注入實驗組(n=5) AFP-USPIO、對照組(n=5) USPIO后0.5小時、1小時、2小時、6小時、12小時、24小時分別行MR增強掃描,測量各時間點病灶、肝臟組織的信號強度,分別計算各時間點的肝臟-腫瘤的對比噪聲比(CNR)及瘤體的信號強度下降百分比(PSIL)。 結(jié)果：MRI平掃T2WI序列上腫瘤組織呈高、稍高信號,部分信號欠均勻。實驗組注射靶向?qū)Ρ葎┖?.5-1小時腫瘤組織T2信號強度下降不明顯,在2-8小時瘤體信號隨著時間延長逐漸減低,瘤體在4h出現(xiàn)一個強化高峰期,并持續(xù)性強化,而12-24小時瘤體信號稍呈上升趨勢。對照組注射靶向?qū)Ρ葎┖蟾鲿r間點腫瘤組織信號未見減低。 實驗組和對照組瘤體的信號測定值差異具有統(tǒng)計學(xué)意義(F=164.5,P0.01),實驗組和對照組的肝臟的信號測定值差異不具有有統(tǒng)計學(xué)意義(F=4.6,P0.01)。實驗組和對照組的腫瘤-肝臟CNR計算值差異具有統(tǒng)計學(xué)意義(F=132.7,P0.01)。 結(jié)論：通過MR掃描動態(tài)觀察AFP-USPIO增強后腫瘤的信號變化,推斷其在腫瘤組織中的分布變化情況。增強后2h-8h瘤體隨著時間推移,呈持續(xù)性強化,4-8h納米粒子在腫瘤組織的聚集達到峰值,12h后逐步退出。
[Abstract]:In our country, the incidence of Hepatocellular (HCC) is the first in the world, and the death rate is second of the death of malignant tumor. Early diagnosis of liver cancer, early diagnosis determines the effect of clinical treatment and prognosis of the patients. Clinical detection methods include serum AFP test and various imaging methods. Alpha-fetoprot Ein, AFP) is a specific marker of HCC, and 80% of the patients may have a rise in alpha fetoprotein. If the anti AFP antibody is linked with some developer or (and) anticancer drugs, the target imaging and targeting therapy of liver cancer can be achieved. It is possible to make imaging diagnosis and clinical treatment simultaneously. This experiment is intended to pass a small dose of two ethyl sub. The MR molecular imaging of the hepatoma model of SD rats induced by nitramine (DENA), the dynamic observation and the pathological comparison, were used to study the specific targeting imaging and distribution of super small superparamagnetic nanoparticles targeting AFP in HCC. This study mainly included two parts.
Part I: establishment of sD rat liver cancer model and MR imaging of targeted AFP-USPIO in SD rat liver cancer.
Objective: to establish a SD rat liver cancer model and AFP antibody labeled USPIO enhanced scan to observe the specific MRI imaging of liver cancer in rats.
Methods: a DENA aqueous solution of 0.1mg/ml was regularly prepared, and 25 male SD rats were given free drinking. After feeding for 14 weeks, 20 rats were selected for the experimental observation of drinking water for 14 weeks. The rats were randomly divided into 2 groups: the experimental group 10 and the control group 10. The rats were first MRI T2WI plain scan, and the experimental group injected AFP-USPI. O (Ba Xiangzu), the control group was injected with USPIO (non target group) 2h after T2WI enhanced scan, observed the signal change of the focus, measured the focus of the lesion, the signal intensity of the liver, calculated the contrast noise ratio (CNR). After the death of rats, the lesion was taken HE, AFP immunization histochemical staining and Prussian blue staining analysis.
Results: MRI scan T2WI showed the high, slightly high signal and mixed signal nodules in the liver, and selected nodules of 3mmm in diameter as the experimental object. The focus state was clear, most of the signals were uniform, high and slightly high. The experimental group, the control group was injected with AFP-USPIO, the 2H enhanced scan after USPIO, and the AFP-USPIO injection of AFP-USPIO. Before and after, the CNR of rat liver tumor was 10.0+2.45 and 4.73 se 2.51 respectively, the difference was statistically significant (P0.001, t=11.23), and the CNR of liver tumor was 9.15+1.24 and 9.96 + 1.63 respectively before and after injection of USPIO, the difference was not statistically significant (P=0.186, t=-1.43).
The pathological results of HE staining showed that all the lesions observed as MRI were hepatocellular carcinoma, and the immunohistochemical results of AFP showed that a large number of AFP. Prussian blue iron staining results in the cytoplasm of the hepatoma cells showed that there were more blue stained granules in the tumor tissue and in the tumor cells in the experimental group, and the blue stained particles in the control group were less.
Conclusion: the model of two ethyl nitrosamine induced SD rat liver cancer is convenient, and the rate of tumor formation is high. The enhanced AFP-USPIO target contrast agent will gather in the rat liver cancer tissue and reduce the signal of tumor tissue, although the contrast noise ratio of tumor liver is reduced, it is helpful for the qualitative diagnosis of liver cancer.
The second part: Experimental Study on the distribution and change of AFP-USPIO in rat liver cancer tissue.
Objective: To study the changes of the time distribution of antibody labeled superparamagnetic iron oxide nanoparticles (AFP-USPIO) in rat liver cancer tissue, and provide theoretical support for the distribution of nanoparticles in the tumor in the next step.
Methods: 10 rat models of SD rat liver cancer were established. The methods and requirements were the same. First, MR T2WI was carried out, then the experimental group (n=5) AFP-USPIO was injected, and the control group (n=5) USPIO after USPIO, 0.5 hours, 1 hours, 2 hours, 6 hours, 12 hours and 24 hours respectively, and measured the focus of each time point, the signal intensity of liver tissue, and calculated the time of each time respectively. The ratio of liver to tumor contrast noise ratio (CNR) and percentage of tumor signal intensity decreased (PSIL).
Results: the tumor tissue in the MRI plain T2WI sequence was high, slightly high signal, and partial signal was not uniform. The decrease of T2 signal intensity in the tumor tissue was not obvious at 0.5-1 hours after the injection of target contrast agent in the experimental group, and the tumor body signal decreased gradually in 2-8 hours with time, and the tumor body appeared a strengthening peak in 4h, and continued to strengthen, and 12-24 hours. Tumor signal showed a slight upward trend. In control group, tumor signal did not decrease at any time after injection of targeted contrast agent.
The difference of the measured values of the signal of the tumor in the experimental group and the control group was statistically significant (F=164.5, P0.01). The difference in the measured values of the liver signal in the experimental group and the control group was not statistically significant (F=4.6, P0.01). The difference in the CNR calculation of the tumor liver of the experimental group and the control group was statistically significant (F=132.7, P0.01).
Conclusion: the changes in the signal of the tumor were observed dynamically by the MR scan, and the distribution of the tumor in the tumor tissue was deduced. After the enhancement, the 2h-8h tumor was continuously strengthened with time, and the aggregation of 4-8h nanoparticles reached the peak in the tumor tissue and gradually withdrew after 12h.

【學(xué)位授予單位】：揚州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R735.7;R445.2

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