【摘要】： 目的研究RGD小分子多肽的合成,RGD多肽水凝膠的制備及其在兔眼結(jié)膜下和前房中的生物相容性。方法通過(guò)標(biāo)準(zhǔn)的9-芴甲氧羰基(FMOC)固相肽合成技術(shù)(SPPS)合成含有RGD序列(精氨酸-甘氨酸-天門(mén)冬氨酸)和N-9-芴甲氧羰基(FMOC)的小分子多肽。將這種超分子多肽溶于蒸餾水,多肽通過(guò)FMOC端基的π-π鍵堆疊作用自組裝成具有三維結(jié)構(gòu)的多肽水凝膠。再把這種RGD多肽水凝膠注射在兔眼結(jié)膜下和前房中,通過(guò)臨床觀察和組織學(xué)研究對(duì)這種新型多肽水凝膠進(jìn)行生物相容性研究。結(jié)果成功合成了含有氨基酸RGD序列(精氨酸-甘氨酸-天門(mén)冬氨酸)的小分子多肽,多肽能通過(guò)自組裝行為得到可注射的RGD多肽水凝膠。RGD多肽水凝膠在兔眼結(jié)膜下和前房中具有良好的生物相容性。結(jié)論RGD多肽水凝膠在兔眼結(jié)膜和前房中生物相容性良好,并具有構(gòu)建眼部植入性的藥物緩釋系統(tǒng)的應(yīng)用潛力,這種新型生物材料及其控釋系統(tǒng)將在眼前節(jié)疾病比如青光眼,虹膜睫狀體炎,角膜病的治療中發(fā)揮作用。 目的一種新型的自組裝多肽水凝膠應(yīng)用于兔眼濾過(guò)術(shù)后結(jié)膜下,使其能達(dá)到降低術(shù)后眼內(nèi)壓(IOP),延長(zhǎng)濾過(guò)泡生存時(shí)間,和促進(jìn)濾過(guò)術(shù)后傷口正常愈合的目的。 方法合成RGD自組裝的多肽水凝膠。進(jìn)行兩組動(dòng)物實(shí)驗(yàn)：1)兔眼行濾過(guò)術(shù)后結(jié)膜下注射多肽水凝膠,不同時(shí)間段處死動(dòng)物,應(yīng)用實(shí)時(shí)定量逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(RT-PCR)測(cè)其濾過(guò)區(qū)域的結(jié)締組織生長(zhǎng)因子(CTGF) mRNA的表達(dá)水平。2)兔眼行濾過(guò)術(shù)后結(jié)膜下注射多肽水凝膠,不同時(shí)間段對(duì)兔眼進(jìn)行眼內(nèi)壓,裂隙燈顯微鏡,超聲生物顯微鏡(UBM)的觀察,并在21天后處死動(dòng)物,對(duì)兔眼進(jìn)行組織學(xué)分析。 結(jié)果結(jié)締組織生長(zhǎng)因子(CTGF) mRNA的表達(dá)水平和眼內(nèi)壓(IOP)在術(shù)后21天均明顯比對(duì)照組低。手術(shù)區(qū)域?yàn)V過(guò)泡形態(tài)和超聲生物顯微鏡(UBM)都提示濾過(guò)泡和濾過(guò)道功能良好。病理組織學(xué)分析提示RGD多肽水凝膠能夠保持兔眼濾過(guò)術(shù)后濾過(guò)泡和濾過(guò)道的形態(tài)結(jié)構(gòu),減少瘢痕形成,并使手術(shù)傷口正常愈合。結(jié)論RGD多肽水凝膠能夠在兔眼濾過(guò)術(shù)后降低結(jié)締組織生長(zhǎng)因子(CTGF) mRNA的表達(dá)水平,延長(zhǎng)濾過(guò)泡生存時(shí)間,抑制濾過(guò)道的瘢痕形成,使眼內(nèi)壓持續(xù)維持較低水平。RGD多肽水凝膠為防止濾過(guò)術(shù)后濾過(guò)道瘢痕化提供了新的治療途徑。 目的構(gòu)建5-FU RGD多肽水凝膠控釋系統(tǒng),研究該控釋系統(tǒng)的體內(nèi)外釋藥情況及抑制兔眼濾過(guò)術(shù)后瘢痕化的作用。方法合成含有RGD序列(精氨酸-甘氨酸-天門(mén)冬氨酸)和N-9-芴甲氧羰基(FMOC)的小分子多肽,多肽通過(guò)自組裝行為得到RGD多肽水凝膠,并構(gòu)建5-FU RGD多肽水凝膠控釋系統(tǒng)。紫外分光光度計(jì)檢測(cè)控釋系統(tǒng)的體外釋藥情況。高效液相(HPLC)色譜儀檢測(cè)5-FU RGD多肽水凝膠控釋系統(tǒng)的兔眼內(nèi)藥代動(dòng)力學(xué)情況。將5-FU RGD多肽水凝膠控釋系統(tǒng)植入于行濾過(guò)術(shù)后的兔眼結(jié)膜下,不同時(shí)間段對(duì)兔眼進(jìn)行眼內(nèi)壓,裂隙燈顯微鏡等臨床觀察,并于3至28天不同時(shí)間段處死動(dòng)物,對(duì)兔眼進(jìn)行組織學(xué)分析。結(jié)果5-FU能從RGD多肽水凝膠控釋系統(tǒng)中緩慢釋放,體內(nèi)外釋藥平穩(wěn)、高效,無(wú)突釋現(xiàn)象。該控釋系統(tǒng)能使兔眼眼內(nèi)壓(IOP)在術(shù)后28天均明顯低于對(duì)照組,明顯減少5-FU對(duì)周?chē)Ｑ劢M織的毒副作用,手術(shù)區(qū)域?yàn)V過(guò)泡形態(tài)提示濾過(guò)泡和濾過(guò)道的濾過(guò)功能良好。病理組織學(xué)分析提示RGD多肽水凝膠能夠保持兔眼濾過(guò)術(shù)后濾過(guò)泡和濾過(guò)道的形態(tài)結(jié)構(gòu),明顯減少α-肌動(dòng)蛋白(α-SMA)的表達(dá)和膠原沉積。結(jié)論5-FU能夠從RGD多肽水凝膠中緩慢釋放,將5-FU RGD自組裝多肽水凝膠應(yīng)用于兔眼濾過(guò)術(shù)后,不僅能夠避免頻繁的結(jié)膜下注射5-FU,明顯減少5-FU對(duì)周?chē)Ｑ劢M織的毒副作用,而且能夠控制濾過(guò)術(shù)后眼內(nèi)壓及抑制濾過(guò)術(shù)后瘢痕化。5-FU RGD多肽水凝膠控釋系統(tǒng)為防止濾過(guò)術(shù)后濾過(guò)道瘢痕化提供了新的治療途徑。
[Abstract]:Objective To study the synthesis of RGD small molecule polypeptide, the preparation of RGD polypeptide hydrogel and the biocompatibility of RGD polypeptide hydrogel in the subconjunctival and anterior chamber of rabbits. Methods Small-molecule polypeptides containing RGD (Arginine-Glycine-Aspartate) and N-9-methoxoxy-base (FMOC) were synthesized by standard 9-methoxyphenyl (FMOC) solid-phase peptide synthesis (SPPS). The super-molecular polypeptide is dissolved in distilled water, and the polypeptide is self-assembled into a polypeptide hydrogel with a three-dimensional structure by a carbon-carbon bond stacking function of the FMOC end group. The RGD polypeptide hydrogel was then injected into the subconjunctival and anterior chamber of the rabbit, and the biological compatibility of the new polypeptide hydrogel was studied by clinical observation and histological study. As a result, a small-molecule polypeptide containing the amino acid RGD sequence (arginine-glycine-aspartic acid) was successfully synthesized, and the polypeptide can be obtained by self-assembly to obtain the injectable RGD polypeptide hydrogel. The RGD polypeptide hydrogel has good biocompatibility in the subconjunctival and anterior chamber of the rabbit. Conclusion The RGD polypeptide hydrogel has good biocompatibility in the conjunctiva and anterior chamber of the rabbit, and has the potential for the application of the sustained-release drug delivery system. The new type of biological material and its controlled-release system will be in the anterior segment of the disease such as glaucoma, iridocyclitis, A role in the treatment of corneal disease. Objective A novel self-assembled polypeptide hydrogel is applied to the conjunctiva of rabbit eye filtering operation, which can reduce the postoperative intraocular pressure (IOP), prolong the survival time of the filtration bubble, and promote the normal healing of the wound after filtration. The purpose of the method is to synthesize the RGD self-assembly The two groups of animal experiments were carried out.1) The subconjunctival injection of the polypeptide hydrogel after the filtration of the rabbit eye line was different. The expression level of connective tissue growth factor (CTGF) mRNA was measured by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). The observation of the slit lamp microscope and the ultrasound biomicroscope (UBM) was performed and the animals were sacrificed after 21 days and the rabbit eyes Histological analysis was performed. The expression level of connective tissue growth factor (CTGF) mRNA and intraocular pressure (IOP) were post-operative 21 days were significantly lower than that in the control group. The function of the bleb and the filter channel is good. The pathological and histological analysis suggests that the RGD polypeptide hydrogel can keep the morphological structure of the bleb and the filter channel after the filtration of the rabbit eye, and reduce the formation of scar. Conclusion The RGD polypeptide hydrogel can decrease the expression level of the connective tissue growth factor (CTGF) mRNA, prolong the survival time of the bleb, and inhibit the formation of the bleeding mark of the filter channel. The internal pressure of the eye was kept low. The RGD polypeptide hydrogel was used to prevent the filtration after the filtration. The purpose of this study is to construct a 5-FU RGD polypeptide hydrogel controlled-release system to study the in vivo release of the controlled-release system. A small-molecule polypeptide containing the RGD sequence (arginine-glycine-aspartic acid) and N-9-bumetacin (FMOC) is synthesized by the method, the RGD polypeptide hydrogel is obtained by self-assembly behavior, and 5 -FU RGD polypeptide hydrogel controlled-release system. In vitro release of a controlled-release system by an altimeter. The 5-FU RGD polypeptide hydrogel was detected by a high performance liquid chromatography (HPLC) chromatograph. A 5-FU RGD polypeptide hydrogel controlled-release system was implanted in the conjunctiva of the rabbit eyes after the line filtration, and the intraocular pressure, slit lamp microscope and other clinical observations were performed on the rabbit eye for different time periods, and different time periods were observed for 3 to 28 days. The results showed that 5-FU was able to release slowly from RGD polypeptide hydrogel controlled-release system. The internal and external drug release was stable, efficient and free of burst. The controlled-release system can make the intraocular pressure (IOP) of the rabbit eyes lower than that of the control group at 28 days after the operation, obviously reduce the toxic and side effect of 5-FU on the surrounding normal eye tissues, The results showed that the filtration function of the bleb and the filter channel was good. The pathological and histological analysis suggested that the RGD polypeptide hydrogel can keep the morphological structure of the bleb and the filter channel after the filtration of the rabbit eye, and obviously reduce the amount of the F-actin. Conclusion 5-FU can be slowly released from the RGD polypeptide hydrogel, and the 5-FU RGD self-assembled polypeptide hydrogel is applied to rabbit eye filtration. The 5-FU RGD polypeptide hydrogel controlled-release system was used to prevent the post-filtration post-filtration.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2010
【分類號(hào)】：R779.6

【共引文獻(xiàn)】

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本文編號(hào)：2440944