本文關(guān)鍵詞：納米靶向FSH受體卵巢早衰大鼠模型建立的相關(guān)研究　出處：《復(fù)旦大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 卵巢早衰 卵泡刺激素 受體 納米粒

【摘要】：卵巢早衰(premature ovarian failure, POF)是指女性40周歲之前發(fā)生的卵巢功能衰竭,以雌激素缺乏(E225pg/mL)及促性腺激素,包括卵泡刺激素(Follicle Stimulating Hormone, FSH)和黃體生成素(Luteinizing Hormone, LH)水平升高(FSH40IU/L,LH30IU/L)為特征,可發(fā)生在月經(jīng)初潮之前或之后,表現(xiàn)為繼發(fā)性閉經(jīng)、月經(jīng)稀發(fā)或絕經(jīng)、不孕。POF是引起婦女不孕和內(nèi)分泌紊亂的常見原因,女性30歲之前的發(fā)病率為0.1%,40歲之前的發(fā)病率為1%。雖然POF的發(fā)病率日趨增高,但其發(fā)病機(jī)制仍不甚明了。為了深入研究POF的發(fā)生及進(jìn)展規(guī)律,探尋安全、有效的治療方法,我們需要建立一種符合病因?qū)W同時又理想、可靠的動物模型。靶向治療具有很強(qiáng)的目的性,可以顯著減少藥物對其它組織的毒副作用。本課題利用FSH多肽片段作為靶向頭基,聯(lián)合納米粒載體,探索構(gòu)建主動靶向大鼠FSH受體的納米系統(tǒng),為進(jìn)一步建立更加有效的FSH受體功能抑制型POF動物模型的提供理論基礎(chǔ)。 第一部分根據(jù)大鼠FSHβ亞基第32-52氨基酸序列成功制備了rFSHβ32-52多肽片段,并將Maleimide-PE-PLA和MPEG-PLA按質(zhì)量比1：9混合,采用復(fù)乳/溶媒蒸發(fā)法制備了載香豆素-6的NP、rFSHβ32-NP,粒徑分別為100和120nm左右,Zeta電位值為-14mV左右。NP表面經(jīng)多肽修飾后粒徑有一定程度的增加,電位值無明顯的變化。 為了驗(yàn)證rFSH32-NP的靶向性,第二部分采用香豆素-6熒光標(biāo)記的NP及rFSHβ32-NP對細(xì)胞進(jìn)行靶向內(nèi)吞能力的定性及定量檢驗(yàn),評價了rFSHβ32-52對大鼠卵巢顆粒細(xì)胞的靶向能力。相較于FSHR陰性的SKOV3細(xì)胞,FSHR陽性的CaOV3和大鼠卵巢顆粒細(xì)胞對rFSHβ32-NP的攝取率顯著增加,并且呈現(xiàn)出一定的時間和濃度依賴性。說明利用rFSHβ32-52多肽片段作為靶向頭基的納米粒對大鼠卵巢顆粒細(xì)胞具有主動靶向的作用。 本研究構(gòu)建的rFSHβ32-NP給藥系統(tǒng),實(shí)現(xiàn)了對大鼠卵巢顆粒細(xì)胞靶向給藥的目的,為進(jìn)一步創(chuàng)建規(guī)范統(tǒng)一化的POF動物模型提供了理論和實(shí)驗(yàn)依據(jù),具有一定的參考價值。
[Abstract]:Premature ovarian failure (premature ovarian failure, POF) refers to the ovarian failure in women 40 years of age before, with the lack of estrogen (E225pg/mL) and gonadotropins, follicle stimulating hormone (Follicle Stimulating, Hormone, FSH) and luteinizing hormone (Luteinizing, Hormone, LH) levels (FSH40IU/L, LH30IU/L) as the characteristic, after can occur in or before menarche, as secondary amenorrhea, oligomenorrhea or menopause, infertility.POF are a common cause of infertility in women and endocrine disorders, the incidence of women before the age of 30 was 0.1%, the incidence rate of 1%. before the age of 40, although the incidence of POF is increasing, but its pathogenesis is still not very clear. For the occurrence and progress of law, in-depth study of POF to explore the safe, effective treatment, we need to establish an accord with the etiology and ideal animal model of targeted therapy has reliable. A strong purpose, can significantly reduce the side effects on other tissues drugs. The issue of the use of FSH polypeptides as targeting ligand, combined with nanoparticles, explore the construction of targeted nano system of the rat FSH receptor, FSH receptor function to further establish a more effective inhibition of the POF animal model theory the foundation.
The first part according to the 32-52 amino acid sequence of rat FSH were successfully synthesized rFSH beta subunit beta 32-52 peptide fragments, and Maleimide-PE-PLA and MPEG-PLA at the mass ratio of 1:9 mixture, using double emulsion / solvent evaporation method to prepare loading coumarin -6 NP, rFSH beta 32-NP, the particle size was about 100 and 120nm respectively, Zeta potential the value is about -14mV.NP after the surface modification with FSHP diameter increased to a certain extent, the potential value of no significant changes.
In order to verify the rFSH32-NP target, the second part of the coumarin fluorescent -6 markers NP and rFSH beta 32-NP qualitative and quantitative test of the ability to swallow inward target cells, and evaluate the rFSH beta 32-52 on rat ovarian granulosa cell targeting ability. Compared to FSHR negative SKOV3 cells of rFSH beta 32-NP uptake ovarian granulosa cells CaOV3 and FSHR positive rate in rats increased significantly, and showed a time and concentration dependent manner. The use of rFSH beta peptide fragments of 32-52 nanoparticles as a targeting ligand has the effect of active targeting on rat ovarian granulosa cells.
The rFSH beta 32-NP delivery system has achieved the goal of targeting the ovarian granulosa cells in rats. It provides a theoretical and experimental basis for further establishing standardized and unified POF animal models, and has a certain reference value.

【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R711.75;R-332

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本文編號：1359685