發(fā)布時(shí)間：2018-05-13 06:17

  本文選題：層狀雙氫氧化物 + 五氟尿嘧啶　； 參考：《第二軍醫(yī)大學(xué)》2016年碩士論文

【摘要】：原發(fā)性肝細(xì)胞癌(Hepatocellular Carcinoma,HCC)位于腫瘤發(fā)病率的第五位,其死亡率在惡性腫瘤中居第三。目前,手術(shù)治療是中早期HCC患者的最佳治療措施。然而,HCC起病較為隱匿,確診時(shí)多屬晚期,已喪失手術(shù)切除機(jī)會(huì)。同時(shí),有臨床數(shù)據(jù)顯示,對(duì)于部分早期手術(shù)切除的患者,術(shù)后復(fù)發(fā)率也居高不下。因此,臨床上化學(xué)藥物治療及介入治療對(duì)原發(fā)性肝癌病人治療具有重要意義。然而,HCC病人大多數(shù)存在不同程度肝硬化,其肝臟解毒功能較差,化療藥物如五氟尿嘧啶(5-Fluorouraci,Fu)等直接暴露于組織,引起較大的毒副作用,同時(shí)隨著經(jīng)典化療藥物的廣泛開(kāi)展,腫瘤多藥耐藥(Multidrug Resistance,MDR)已經(jīng)成為腫瘤病人化學(xué)藥物治療的巨大障礙。文獻(xiàn)調(diào)研發(fā)現(xiàn),多種機(jī)制參與腫瘤的多耐藥,如細(xì)胞膜的糖蛋白機(jī)制,腫瘤的抗凋亡基因上調(diào)以及自噬等。因此,如何解決腫瘤耐藥已經(jīng)成為腫瘤病人化療成功與否的關(guān)鍵因素。常見(jiàn)的措施包括增大藥物的劑量從而達(dá)到藥物有效濃度,但卻帶來(lái)了更大的毒副作用風(fēng)險(xiǎn)。近年來(lái),化療藥物的聯(lián)合療法為腫瘤耐藥病人帶來(lái)希望,可以通過(guò)發(fā)揮不同藥物的功能協(xié)同殺傷腫瘤,但多種化療藥的同時(shí)作用存在諸多不確定因素,聯(lián)合藥物本身也可能帶來(lái)更大的毒副作用。姜黃素(Curcumin,Cur)是食物來(lái)源的傳統(tǒng)型藥物,具有廉價(jià)、無(wú)毒等諸多優(yōu)點(diǎn)。研究表明,姜黃素具有抗炎、抗氧化、抗腫瘤等多重作用,文獻(xiàn)報(bào)道顯示,化療藥物與姜黃素聯(lián)合能夠有效逆轉(zhuǎn)腫瘤多耐藥。然而,姜黃素具有水溶性差,體內(nèi)代謝快等不足,如何充分發(fā)揮姜黃素應(yīng)有的強(qiáng)大功能是實(shí)現(xiàn)其抑制耐藥效果的關(guān)鍵。近年來(lái)納米技術(shù)的發(fā)展為腫瘤藥物劑型提供新方向,納米技術(shù)具有以下優(yōu)點(diǎn):納米載體的pH敏感性、溫度敏感性可以實(shí)現(xiàn)藥物的控釋,從而提高藥物利用度和有效降低藥物因直接暴露帶來(lái)的毒副作用;納米載體的溫敏功能及不同配體的修飾可以實(shí)現(xiàn)被動(dòng)及主動(dòng)靶向;納米技術(shù)的雞尾酒療法不僅僅局限于兩種藥物混合使用,可以實(shí)現(xiàn)化療藥物的時(shí)序性、協(xié)同性作用。層狀雙氫氧化物(Layered hydroxide,LDH)是一類(lèi)陰離子型黏土,它具有雙層板狀結(jié)構(gòu),LDH板層外帶正電荷,且富含大量氫氧根,便于形成氫鍵而載藥。板間帶負(fù)電荷,可用于陰離子物質(zhì)交換,可負(fù)載電離為陰離子的藥物以及小分子的核酸。LDH具有無(wú)毒、高載藥量和pH敏感性等優(yōu)點(diǎn)而引起廣泛關(guān)注,是不同腫瘤藥物的良好載體。本課題中,我們利用LDH層間負(fù)載Fu以及Cur構(gòu)建納米混合劑型,通過(guò)Bel 7402/Fu肝癌耐藥細(xì)胞為模型,研究納米LDH劑型對(duì)于氟尿嘧啶耐藥的肝癌腫瘤作用。細(xì)胞水平首先考察了 LDH-Fu-Cur對(duì)于正常肝臟細(xì)胞LO_2及Bel 7402/Fu的增殖抑制作用。采用流式凋亡手段檢測(cè)LDH-Fu-Cur的促凋亡作用,并采用Western blot技術(shù)檢測(cè)凋亡相關(guān)機(jī)制。通過(guò)共聚焦檢測(cè)耐藥細(xì)胞對(duì)于納米藥物的吞噬作用以及納米藥物引起的自噬,利用自噬工具藥檢測(cè)自噬在LDH-Fu-Cur抗腫瘤中的效應(yīng)。最后采取小鼠抑瘤實(shí)驗(yàn)檢測(cè)LDH-Fu-Cur在體內(nèi)的抑瘤效果。系統(tǒng)研究顯示:我們成功制備具有高載藥量且生物安全性良好的LDH共載藥體系,紫外分光光度計(jì)檢測(cè)納米體系具有較高的載藥量,LDH-Fu中Fu載藥量為20.3%±0.4%,LDH-Fu-Cur 中 Fu 載藥量為 19.7%±0.2%,Cur 的載藥量 15.7%±0.6%,符合實(shí)驗(yàn)預(yù)期結(jié)果,能夠滿(mǎn)足實(shí)驗(yàn)要求。實(shí)驗(yàn)還提示LDH-Fu-Cur對(duì)于正常肝臟LO_2細(xì)胞沒(méi)有明顯毒副作用,而對(duì)于Bel 7402/Fu腫瘤細(xì)胞具有明顯的抑制作用。流式細(xì)胞技術(shù)及Western blot進(jìn)一實(shí)驗(yàn)證實(shí)LDH-Fu-Cur能夠較單純姜黃素組具有明顯的促Bel 7402/Fu細(xì)胞凋亡效應(yīng)。有意思的是:本課題研究發(fā)現(xiàn)自噬協(xié)同參與了在體系對(duì)肝癌細(xì)胞的抑制作用。雖然細(xì)胞自噬是一把"雙刃劍",我們的實(shí)驗(yàn)還證實(shí)LDH-Fu-Cur能夠啟動(dòng)細(xì)胞自噬,而這種自噬是一種促進(jìn)細(xì)胞死亡的協(xié)同殺傷性自噬。最后,通過(guò)構(gòu)建肝癌Bel 7402/Fu裸鼠異種瘤移植模型,觀(guān)察LDH-Fu-Cur組、其他藥物組以及PBS對(duì)照組發(fā)現(xiàn),LDH-Fu-Cur能夠明顯抑制Bel 7402/Fu細(xì)胞在裸鼠體內(nèi)生長(zhǎng),提示LDH-Fu-Cur可作為抑制氟尿嘧啶耐藥肝癌細(xì)胞的有效藥物。綜上所述,通過(guò)本課題開(kāi)展,我們制備了結(jié)構(gòu)良好、生物安全的LDH-Fu-Cur納米藥物,實(shí)現(xiàn)氟尿嘧啶和姜黃素共載,新的納米藥物在體內(nèi)、體外均展現(xiàn)處良好抑制肝癌氟尿嘧啶耐藥細(xì)胞,為肝癌抗腫瘤耐藥藥物的研發(fā)提供新思路。
[Abstract]:Hepatocellular Carcinoma (HCC) is located in fifth of the incidence of tumor, and its mortality rate is third in the malignant tumor. At present, surgical treatment is the best treatment for the early middle HCC patients. However, the onset of HCC is more concealed, and the diagnosis is late and has lost the chance of surgical resection. The rate of postoperative recurrence is also high in some patients with early surgical resection. Therefore, the clinical chemotherapeutic and interventional therapy is of great significance for the treatment of primary liver cancer patients. However, most of HCC patients have different degrees of liver cirrhosis, their liver detoxification function is poor, chemotherapy drugs such as five fluorouracil (5-Fluorouraci, Fu), and so on. Direct exposure to tissue causes large toxic and side effects, and with the extensive development of classical chemotherapeutic drugs, Multidrug Resistance (MDR) has become a huge obstacle to the treatment of chemical drugs in cancer patients. Therefore, how to solve the drug resistance has become a key factor in the success of chemotherapy in cancer patients. The common measures include increasing the dose of the drug to achieve the effective concentration of the drug, but it brings a greater risk of toxic and side effects. In recent years, the combined therapy of chemotherapeutic drugs has been used as a drug resistant patient. It brings hope that the functions of different drugs can be combined to kill the tumor, but there are many uncertainties in the simultaneous action of various chemotherapeutic agents. The combined drug itself may also bring more toxic and side effects. Curcumin (Cur) is a traditional drug of food source, which has many advantages such as cheap, non-toxic and many other advantages. Studies show Jiang Huang It has been reported that the combination of chemotherapeutic drugs and curcumin can effectively reverse the multidrug resistance of tumor. However, curcumin has poor water solubility and fast metabolism in the body. How to fully exert the strong work energy of curcumin is the key to its inhibition of drug resistance. The development of technology provides a new direction for drug dosage forms. Nanotechnology has the following advantages: pH sensitivity of nanoscale, temperature sensitivity can be used to control drug release, so as to improve drug utilization and effectively reduce the toxic and side effects caused by direct exposure to drugs; the temperature sensitive function of nanoscale and the modification of different ligands can be realized. The nanotechnology's cocktail therapy is not only limited to the mixed use of two drugs, and can realize the timing and synergistic effect of the chemotherapeutic drugs. The layered double hydroxides (Layered hydroxide, LDH) are a class of anionic clay. It has a double layered structure, a positive charge of the outer band of the LDH plate and a large amount of hydrogen and oxygen. Root, which facilitates the formation of hydrogen bonds and carrying drugs. The interplate with negative charge, can be used for anion exchange, can be loaded into anions, and the nucleic acid.LDH of small molecules has non-toxic, high drug loading and pH sensitivity. It is a good carrier for different tumor drugs. In this subject, we use LDH interlayer load Fu to And Cur construction of nano mixture type, through Bel 7402/Fu hepatoma resistant cells as model, study the effect of nano LDH on fluorouracil resistant cancer of liver cancer. Cell level first investigated the proliferation inhibition effect of LDH-Fu-Cur on normal liver cells LO_2 and Bel 7402/Fu. Flow apoptosis was used to detect the apoptosis of LDH-Fu-Cur. The apoptosis related mechanism was detected by Western blot technique. The effect of autophagy on the anti-tumor effect of autophagy in LDH-Fu-Cur was detected by confocal microscopy, and autophagy was used to detect the effect of autophagy on the anti-tumor effect of LDH-Fu-Cur. Finally, the tumor suppressor test was used to detect the tumor inhibition effect of LDH-Fu-Cur in the body. The systematic study shows that we have successfully prepared the LDH co carrier system with high load and good biological safety. The UV spectrophotometer has high drug loading, the Fu load of Fu in LDH-Fu is 20.3% + 0.4%, the dosage of Fu in LDH-Fu-Cur is 19.7% + 0.2%, and the drug loading of Cur is 15.7% + 0.6%, which conforms to the expected result of the experiment. The experiment also showed that LDH-Fu-Cur had no obvious toxic and side effects on normal liver LO_2 cells, but had obvious inhibitory effect on Bel 7402/Fu tumor cells. Flow cytometry and Western blot experiment confirmed that LDH-Fu-Cur could significantly promote the apoptosis effect of Bel 7402/Fu cell than the simple curcumin group. It is interesting: the study found that autophagy participates in the inhibitory effect of autophagy on liver cancer cells. Although cell autophagy is a "double-edged sword", our experiment also confirms that LDH-Fu-Cur can initiate autophagy, which is a synergistic autophagy that promotes cell death. Finally, the construction of Bel 7402/Fu for liver cancer is constructed. The xenograft model of nude mice, observation group LDH-Fu-Cur, other drug groups and PBS control group, found that LDH-Fu-Cur can obviously inhibit the growth of Bel 7402/Fu cells in nude mice, suggesting that LDH-Fu-Cur can be used as an effective drug to inhibit the fluorouracil resistant liver cancer cells. The LDH-Fu-Cur nanoscale drug is safe to carry out the co loading of fluorouracil and curcumin. The new nanoscale drugs in the body show good inhibition of fluorouracil resistant cells in vitro, and provide new ideas for the research and development of anti tumor drug resistance drugs for liver cancer.

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R735.7

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 魯純素,王夔;鉑尿嘧啶蘭形成反應(yīng)動(dòng)力學(xué)的研究[J];北京醫(yī)學(xué)院學(xué)報(bào);1981年03期

2 黃紅兵,黃民,李蘇,廖海,劉韜,余更生,姜文奇;內(nèi)源性尿嘧啶類(lèi)物質(zhì)比值在5-氟尿嘧啶化療中毒性預(yù)測(cè)價(jià)值的臨床研究[J];中國(guó)臨床藥理學(xué)雜志;2002年01期

3 岳瑤;5-氟尿嘧啶的耐藥與逆轉(zhuǎn)研究進(jìn)展[J];臨床腫瘤學(xué)雜志;2003年05期

4 閆美興,初曉,高蕾;氟尿嘧啶的臨床新用途[J];中國(guó)臨床醫(yī)生;2003年09期

5 魯麗峰;超聲介入注射氟尿嘧啶合劑治療卵巢漿液性囊腺瘤[J];中國(guó)煤炭工業(yè)醫(yī)學(xué)雜志;2003年09期

6 陳向紅;許健健;喻平;周麗春;竇紅漫;孫昕;;植入用緩釋氟尿嘧啶犬皮下局部毒性實(shí)驗(yàn)[J];安徽醫(yī)藥;2006年06期

7 Yuge S.;SantosGodoy D.A.;Coelho deMeloM.C. ;李曉莉;;局部外用氟尿嘧啶治療邊緣性離心性角化棘皮瘤療效報(bào)道[J];世界核心醫(yī)學(xué)期刊文摘(皮膚病學(xué)分冊(cè));2006年08期

8 金香;王艷東;;5-氟尿嘧啶在青光眼濾過(guò)性手術(shù)過(guò)程中的應(yīng)用[J];遼寧醫(yī)學(xué)雜志;2006年06期

9 郝愛(ài)軍;鄧英杰;陳長(zhǎng)蘭;陳立江;;氟尿嘧啶乙酸的口服藥動(dòng)學(xué)研究[J];遼寧大學(xué)學(xué)報(bào)(自然科學(xué)版);2008年02期

10 李沛雨;劉娜;李榮;衛(wèi)勃;;緩釋氟尿嘧啶輔助治療腹膜假性粘液瘤的安全性研究和短期隨訪(fǎng)[J];中國(guó)醫(yī)藥導(dǎo)刊;2009年02期

相關(guān)會(huì)議論文 前10條

1 張瓊瑤;趙月萍;葉勇;;磁性5-氟尿嘧啶核殼納米粒子的制備及其載藥、釋藥性能[A];湖北省化學(xué)化工學(xué)會(huì)第十一屆分析化學(xué)專(zhuān)業(yè)年會(huì)論文集[C];2007年

2 張二變;郝玉萍;;氟尿嘧啶在兩種不同途徑化療中的臨床效果觀(guān)察[A];山西省抗癌協(xié)會(huì)第六屆腫瘤學(xué)術(shù)交流會(huì)論文匯編[C];2003年

3 韓士田;劉彥欽;扈睛;;卟啉-5-氟尿嘧啶新化合物的合成[A];中國(guó)化學(xué)會(huì)第四屆有機(jī)化學(xué)學(xué)術(shù)會(huì)議論文集（上冊(cè)）[C];2005年

4 陳葛裙;;5-氟尿嘧啶在皮膚科的應(yīng)用[A];2003中國(guó)中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會(huì)議論文匯編[C];2003年

5 李振亞;鄭連義;;氟尿嘧啶微球的制備研究[A];第一屆全國(guó)化學(xué)工程與生物化工年會(huì)論文摘要集(下)[C];2004年

6 劉莉;過(guò)瑋;宋俊峰;;伏安法測(cè)定尿嘧啶[A];中國(guó)化學(xué)會(huì)第十四屆有機(jī)分析及生物分析學(xué)術(shù)研討會(huì)會(huì)議論文摘要集[C];2007年

7 高君偉;郭彥琨;劉皋林;;二氫嘧啶脫氫酶基因多態(tài)性對(duì)氟尿嘧啶化療毒性的影響[A];2013年中國(guó)臨床藥學(xué)學(xué)術(shù)年會(huì)暨第九屆臨床藥師論壇論文集[C];2013年

8 胥彬;喻平;許健健;;氟尿嘧啶植入劑與腫瘤器官靶向治療的研究[A];2009醫(yī)學(xué)前沿論壇暨第十一屆全國(guó)腫瘤藥理與化療學(xué)術(shù)會(huì)議論文集[C];2009年

9 代小娟;劉坤輝;蘇紅梅;;5-碘代尿嘧啶光化學(xué)反應(yīng)實(shí)驗(yàn)及理論研究[A];中國(guó)化學(xué)會(huì)第29屆學(xué)術(shù)年會(huì)摘要集——第39分會(huì)：化學(xué)動(dòng)力學(xué)[C];2014年

10 胡冰;莊建生;季楚舒;林新民;胡長(zhǎng)路;江豐收;徐騰云;陳曼萍;丁健;何義富;孫玉蓓;張萬(wàn)亨;;皮下植入用緩釋氟尿嘧啶同靜脈用氟尿嘧啶治療食道、賁門(mén)癌近期臨床療效觀(guān)察[A];安徽省抗癌協(xié)會(huì)第四次代表大會(huì)暨乳腺癌、肺癌專(zhuān)業(yè)委員會(huì)成立會(huì)議、安徽省腫瘤防治進(jìn)展學(xué)術(shù)研討會(huì)論文匯編[C];2001年

相關(guān)重要報(bào)紙文章 前5條

1 孫華君;直腸癌患者術(shù)后5－氟尿嘧啶聯(lián)合化療再配合放療的Ⅲ期臨床試驗(yàn)效果觀(guān)察[N];醫(yī)藥經(jīng)濟(jì)報(bào);2006年

2 是明啟;抗癌之外展身手[N];醫(yī)藥經(jīng)濟(jì)報(bào);2001年

3 ;晚期胃癌化療 國(guó)內(nèi)方案少依據(jù)[N];中國(guó)醫(yī)藥報(bào);2005年

4 ;在進(jìn)展期難治性結(jié)直腸癌患者中進(jìn)行貝伐單抗聯(lián)合5－氟尿嘧啶＋亞葉酸的Ⅱ期多中心臨床試驗(yàn)[N];醫(yī)藥經(jīng)濟(jì)報(bào);2006年

5 記者　 張顯峰 陳磊;我科學(xué)家成功解析出H/ACA復(fù)合物完整結(jié)構(gòu)[N];科技日?qǐng)?bào);2006年

相關(guān)博士學(xué)位論文 前7條

1 廖鉑;伏立諾他增強(qiáng)肝癌細(xì)胞對(duì)5-氟尿嘧啶及奧沙利鉑的化療敏感性[D];華中科技大學(xué);2015年

2 華心仲;基于飛秒瞬態(tài)吸收光譜的尿嘧啶激發(fā)態(tài)動(dòng)力學(xué)研究[D];中國(guó)科學(xué)院研究生院(武漢物理與數(shù)學(xué)研究所);2016年

3 熊靜;5-氟尿嘧啶氨基酸類(lèi)化合物的合成及抗癌活性[D];大連理工大學(xué);2009年

4 郭慶海;5-氟尿嘧啶食道支架的制備和體外研究[D];上海交通大學(xué);2007年

5 王晶;吲哚美辛5-氟尿嘧啶甲酯的合成及其吸干乳劑的研究[D];沈陽(yáng)藥科大學(xué);2007年

6 于碩;癌因性疲乏病因機(jī)制的初步研究[D];河北醫(yī)科大學(xué);2010年

7 楊鴿;抗氧化劑調(diào)控Src激酶活性減弱5-氟尿嘧啶誘導(dǎo)的人結(jié)腸癌細(xì)胞凋亡的機(jī)制研究[D];鄭州大學(xué);2014年

相關(guān)碩士學(xué)位論文 前10條

1 馬希中;莪術(shù)醇聯(lián)合氟尿嘧啶對(duì)裸鼠胃癌移植瘤生長(zhǎng)及PCNA和P27表達(dá)影響的研究[D];河北醫(yī)科大學(xué);2015年

2 楊楊;消癌平注射液聯(lián)合氟尿嘧啶及絲裂霉素對(duì)肺癌A549細(xì)胞周期、凋亡及相應(yīng)基因表達(dá)的影響[D];河北醫(yī)科大學(xué);2015年

3 馬存;熊果酸及其衍生物與5-氟尿嘧啶衍生物的鍵聯(lián)及抗癌活性研究[D];廣東藥學(xué)院;2015年

4 孫崇彬;一種含氟大分子前藥的制備及其釋放行為的研究[D];濟(jì)南大學(xué);2015年

5 梁基韻;大蒜素增加肝癌細(xì)胞對(duì)氟尿嘧啶敏感性的初步機(jī)制研究[D];南方醫(yī)科大學(xué);2013年

6 廖年生;5-氟尿嘧啶與他米巴羅汀前藥的設(shè)計(jì)、合成及活性研究[D];山東大學(xué);2016年

7 董馨文;Yg美拉唑和5-氟尿嘧啶合用對(duì)人胃癌細(xì)胞的抗增殖、遷移作用研究[D];山東大學(xué);2016年

8 葉志強(qiáng);Src蛋白激酶抑制劑對(duì)5-氟尿嘧啶誘導(dǎo)的結(jié)腸癌細(xì)胞凋亡的影響及意義[D];鄭州大學(xué);2016年

9 董偉;納米層狀雙氫氧化物共負(fù)載體系的制備及其抑制肝癌氟尿嘧啶耐藥的協(xié)同效應(yīng)機(jī)制研究[D];第二軍醫(yī)大學(xué);2016年

10 李大帥;5-氟尿嘧啶N1取代類(lèi)衍生物的制備與研究[D];溫州大學(xué);2014年

，

本文編號(hào)：1882025