姜黃素殼聚糖脂質(zhì)體治療兔角膜堿燒傷的研究
發(fā)布時(shí)間：2018-10-30 07:00
【摘要】：目的:制備姜黃素殼聚糖脂質(zhì)體,檢測(cè)該藥在眼部應(yīng)用安全基礎(chǔ)上,建立兔角膜堿燒傷模型,評(píng)價(jià)該脂質(zhì)體對(duì)兔角膜堿燒傷的治療作用,并初步探討其作用機(jī)制,為姜黃素殼聚糖脂質(zhì)體滴眼液的臨床應(yīng)用提供實(shí)驗(yàn)依據(jù)。方法:(1)姜黃素殼聚糖脂質(zhì)體的制備和評(píng)價(jià)。采用薄膜分散法制備姜黃素殼聚糖脂質(zhì)體(curcumin-loaded chitosan liposomes,CCLP),測(cè)定其粒徑和Zeta電位,離心法測(cè)定其包封率。(2)姜黃素殼聚糖脂質(zhì)體眼部應(yīng)用安全性評(píng)價(jià)。6只兔隨機(jī)分為姜黃素殼聚糖脂質(zhì)體組和生理鹽水組,每組3只,右眼為實(shí)驗(yàn)眼。姜黃素殼聚糖脂質(zhì)體組滴姜黃素殼聚糖脂質(zhì)體滴眼液,生理鹽水組滴生理鹽水(0.9%Nacl),4次/天,連續(xù)給藥14天,通過(guò)肉眼觀察兔子反應(yīng)、裂隙燈微觀觀察、采用Draize眼刺激性實(shí)驗(yàn)評(píng)分、病理切片,評(píng)價(jià)姜黃素殼聚糖脂質(zhì)體滴眼液對(duì)正常兔眼有無(wú)明顯毒性。(3)姜黃素殼聚糖脂質(zhì)體治療兔角膜堿燒傷的研究。35只新西蘭大白兔隨機(jī)分為:正常對(duì)照組3只、生理鹽水組8只、空白殼聚糖包衣脂質(zhì)體組8只、地塞米松組8只和姜黃素殼聚糖脂質(zhì)體組8只,除正常對(duì)照組外,均選用右眼建立角膜堿燒傷模型,各組分別給予相應(yīng)藥物治療,4次/天,連續(xù)給藥14天。裂隙燈觀察角膜新生血管(corneal neovascularization,CNV)生長(zhǎng)情況及角上皮愈合情況,并用裂隙燈進(jìn)行眼前節(jié)照相;分析CNV的長(zhǎng)度及面積,角膜上皮愈合率。于堿燒傷后1、4、7、14天,取兔房水檢測(cè)白介素-8(interleukin-8,IL-8)的含量。堿燒傷后7、14和28天隨機(jī)抽取各組兔取角膜,行病理切片觀察角膜形態(tài);免疫組化法觀察血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)的表達(dá)情況。結(jié)果:(1)姜黃素殼聚糖脂質(zhì)體的性質(zhì)。成功制備出姜黃素殼聚糖脂質(zhì)體滴眼液,測(cè)得平均粒徑為96.6±14.7 nm,平均Zeta電位為58.8±2.3 m V,平均包封率為51.41±1.05%,符合眼用制劑要求,可作為滴眼液用于動(dòng)物實(shí)驗(yàn)。(2)姜黃素殼聚糖脂質(zhì)體安全性好。短期局部應(yīng)用姜黃素殼聚糖脂質(zhì)體滴眼后兔子無(wú)明顯撓抓眼部情況;裂隙燈檢查,結(jié)膜、角膜透明,虹膜無(wú)滲出,晶體透明;兩組角膜、虹膜及結(jié)膜Draize評(píng)分比較無(wú)差異(p0.05);病理切片示角膜無(wú)炎性細(xì)胞。(3)姜黃素殼聚糖脂質(zhì)體、地塞米松(dexamethasone,DX)和生理鹽水組相比均能有效抑制CNV的生長(zhǎng)(p0.05),空白殼聚糖包衣脂質(zhì)體組和生理鹽水組CNV面積和長(zhǎng)度沒(méi)有顯著差異(p0.05)。與地塞米松組相比,在治療早期,姜黃素殼聚糖脂質(zhì)體組的CNV長(zhǎng)度和面積大(p0.05);在治療后期,地塞米松組和姜黃素殼聚糖脂質(zhì)體組的CNV長(zhǎng)度和面積無(wú)統(tǒng)計(jì)學(xué)差異(p0.05)。姜黃素殼聚糖脂質(zhì)體組與生理鹽水組相比,能促進(jìn)角膜上皮恢復(fù)(p0.05),減輕炎癥,有效降低角膜組織中VEGF的表達(dá)(p0.05),降低房水中IL-8的表達(dá)(p0.05)。結(jié)論:姜黃素殼聚糖脂質(zhì)體藥物包封率較高,安全性好,可明顯減輕兔角膜堿燒傷炎癥,抑制角膜新生血管生長(zhǎng),促進(jìn)角膜上皮修復(fù),降低角膜VEGF表達(dá)和房水中IL-8表達(dá)。姜黃素殼聚糖脂質(zhì)體有望成為新型的治療角膜堿燒傷的局部用藥制劑,為角膜堿燒傷治療提供新思路。
[Abstract]:Objective: To prepare curcumin chitosan liposome, and to test the effect of the liposome on rabbit corneal alkali burn, and to discuss its mechanism of action. It provides experimental basis for clinical application of curcumin chitosan liposome eye drops. Methods: (1) Preparation and evaluation of curcumin chitosan liposomes. The entrapment efficiency of curcumin chitosan liposomes (CCLP) was determined by thin-film dispersion method and its particle size and Zeta potential were measured. (2) The safety evaluation of curcumin chitosan liposome eyes was evaluated. Six rabbits were randomly divided into the curcumin chitosan liposome group and the normal saline group, each group had 3 eyes, and the right eye was the experimental eye. The curcumin chitosan liposome group drops curcumin chitosan liposome eye drops, physiological saline group drops normal saline (0.9% Nacl), 4 times per day, continuously administrated for 14 days, the rabbit reaction and slit lamp microscopic observation are observed by naked eyes, the Draize eye irritation experiment score and pathological section are adopted, To evaluate the toxicity of curcumin chitosan liposome eye drops to the eyes of normal rabbits. (3) The effects of curcumin chitosan liposome on corneal alkali burn in rabbits were studied. Thirty-five New Zealand white rabbits were randomly divided into three groups: normal control group (n = 3), saline group (8), blank chitosan coated liposome group (8), dexamethasone group (8) and curcumin chitosan liposome group (8). In addition to the normal control group, the right eye was used to establish the corneal alkali burn model, each group was given the corresponding drug treatment, 4 times/ day, and the continuous administration was 14 days. slit lamp was used to observe the growth of corneal neovascularization and angle epithelial healing, and the anterior segment was photographed with slit lamp; the length and area of the cornea were analyzed, and the corneal epithelial healing rate was analyzed. The levels of interleukin-8 (IL-8) and interleukin-8 (IL-8) were detected on 1, 4, 7 and 14 days after alkali burn. The corneal morphology was observed on 7, 14 and 28 days after alkali burn, and the expression of vascular endothelial growth factor (VEGF) was observed by immunohistochemistry. Results: (1) The properties of curcumin chitosan liposomes were studied. In this paper, curcumin chitosan liposome eye drops were successfully prepared. The average particle size was 96. 6, 14. 7 nm, the average Zeta potential was 58. 8, 2. 3 m V, the average entrapment efficiency was 51. 41% 1. 05%, which was consistent with the requirements of eye preparations and could be used as eye drops for animal experiments. (2) curcumin chitosan liposome has good safety. In short-term topical application of curcumin chitosan liposome, the rabbit did not scratch the eyes obviously; slit lamp examination, conjunctival, cornea transparency, iris did not exude, crystal clear; two groups of cornea, iris and conjunctival draft score were not different (P0.05); pathological section showed no inflammatory cells of cornea. (3) There was no significant difference between curcumin chitosan liposome, dexamethasone (dexamethasone, DX) and saline group (P0.05). Compared with the dexamethasone group, in the early stage of treatment, there was no significant difference in length and area between the dexamethasone group and the curcumin chitosan liposome group (P0.05). Compared with normal saline group, curcumin chitosan liposome group can promote corneal epithelial recovery (A0.05), reduce inflammation, effectively reduce the expression of VEGF in corneal tissue (P0.05), and decrease the expression of IL-8 in the water. Conclusion: The entrapment efficiency of curcumin chitosan liposome is high, the safety is good, the corneal alkali burn inflammation can be obviously reduced, corneal neovascularization can be inhibited, corneal epithelium repair is promoted, corneal VEGF expression and IL-8 expression in house water can be reduced. The curcumin chitosan liposome is expected to be a novel topical medicine preparation for treating corneal alkali burn, and provides a new idea for the treatment of corneal alkali burn.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R772.2
本文編號(hào)：2299275
[Abstract]:Objective: To prepare curcumin chitosan liposome, and to test the effect of the liposome on rabbit corneal alkali burn, and to discuss its mechanism of action. It provides experimental basis for clinical application of curcumin chitosan liposome eye drops. Methods: (1) Preparation and evaluation of curcumin chitosan liposomes. The entrapment efficiency of curcumin chitosan liposomes (CCLP) was determined by thin-film dispersion method and its particle size and Zeta potential were measured. (2) The safety evaluation of curcumin chitosan liposome eyes was evaluated. Six rabbits were randomly divided into the curcumin chitosan liposome group and the normal saline group, each group had 3 eyes, and the right eye was the experimental eye. The curcumin chitosan liposome group drops curcumin chitosan liposome eye drops, physiological saline group drops normal saline (0.9% Nacl), 4 times per day, continuously administrated for 14 days, the rabbit reaction and slit lamp microscopic observation are observed by naked eyes, the Draize eye irritation experiment score and pathological section are adopted, To evaluate the toxicity of curcumin chitosan liposome eye drops to the eyes of normal rabbits. (3) The effects of curcumin chitosan liposome on corneal alkali burn in rabbits were studied. Thirty-five New Zealand white rabbits were randomly divided into three groups: normal control group (n = 3), saline group (8), blank chitosan coated liposome group (8), dexamethasone group (8) and curcumin chitosan liposome group (8). In addition to the normal control group, the right eye was used to establish the corneal alkali burn model, each group was given the corresponding drug treatment, 4 times/ day, and the continuous administration was 14 days. slit lamp was used to observe the growth of corneal neovascularization and angle epithelial healing, and the anterior segment was photographed with slit lamp; the length and area of the cornea were analyzed, and the corneal epithelial healing rate was analyzed. The levels of interleukin-8 (IL-8) and interleukin-8 (IL-8) were detected on 1, 4, 7 and 14 days after alkali burn. The corneal morphology was observed on 7, 14 and 28 days after alkali burn, and the expression of vascular endothelial growth factor (VEGF) was observed by immunohistochemistry. Results: (1) The properties of curcumin chitosan liposomes were studied. In this paper, curcumin chitosan liposome eye drops were successfully prepared. The average particle size was 96. 6, 14. 7 nm, the average Zeta potential was 58. 8, 2. 3 m V, the average entrapment efficiency was 51. 41% 1. 05%, which was consistent with the requirements of eye preparations and could be used as eye drops for animal experiments. (2) curcumin chitosan liposome has good safety. In short-term topical application of curcumin chitosan liposome, the rabbit did not scratch the eyes obviously; slit lamp examination, conjunctival, cornea transparency, iris did not exude, crystal clear; two groups of cornea, iris and conjunctival draft score were not different (P0.05); pathological section showed no inflammatory cells of cornea. (3) There was no significant difference between curcumin chitosan liposome, dexamethasone (dexamethasone, DX) and saline group (P0.05). Compared with the dexamethasone group, in the early stage of treatment, there was no significant difference in length and area between the dexamethasone group and the curcumin chitosan liposome group (P0.05). Compared with normal saline group, curcumin chitosan liposome group can promote corneal epithelial recovery (A0.05), reduce inflammation, effectively reduce the expression of VEGF in corneal tissue (P0.05), and decrease the expression of IL-8 in the water. Conclusion: The entrapment efficiency of curcumin chitosan liposome is high, the safety is good, the corneal alkali burn inflammation can be obviously reduced, corneal neovascularization can be inhibited, corneal epithelium repair is promoted, corneal VEGF expression and IL-8 expression in house water can be reduced. The curcumin chitosan liposome is expected to be a novel topical medicine preparation for treating corneal alkali burn, and provides a new idea for the treatment of corneal alkali burn.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R772.2
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 王群;吳志鴻;;魚(yú)腥草滴眼液眼局部應(yīng)用的安全性研究[J];環(huán)球中醫(yī)藥;2014年09期
2 郝廷靜;;加溫后的生理鹽水結(jié)膜囊沖洗液的臨床應(yīng)用[J];局解手術(shù)學(xué)雜志;2014年01期
3 趙偉;周瑾;郭夢(mèng)翔;項(xiàng)道滿;;地塞米松對(duì)堿燒傷后大鼠角膜核轉(zhuǎn)錄因子-κB活化的影響[J];眼科新進(jìn)展;2013年07期
4 趙琳;;102例眼部化學(xué)燒傷的急救治療分析[J];吉林醫(yī)學(xué);2012年11期
5 徐正同;;60例眼部化學(xué)燒傷綜合治療的臨床分析[J];中國(guó)臨床研究;2011年01期
6 孫慶雪;邵偉;黃桂華;;脂質(zhì)體制備方法的選擇[J];中成藥;2010年08期
7 蘇鐵芬,Bettina Tilton,袁永輝,李娜萍,Martin Schilling,吳人亮;IL-8在食管癌中表達(dá)對(duì)腫瘤血管形成及患者預(yù)后的影響[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2005年04期
8 張曉承,王智彪,吳群;中和劑持續(xù)滴眼沖洗治療眼酸堿化學(xué)傷[J];眼外傷職業(yè)眼病雜志.附眼科手術(shù);2004年01期
9 鄒毓韜,邱孝芝;兔眼部堿燒傷后角膜組織的pH變化及早期沖洗的影響[J];眼科研究;2000年06期
10 尹瀾;銀頌;皮裕t ;何潤(rùn)西;;眼堿燒傷多次前房沖洗后仍發(fā)現(xiàn)房水pH值異常一例[J];中華眼科雜志;2010年11期
相關(guān)博士學(xué)位論文 前1條
1 石磊;靶向Bcl-2小分子化合物納米懸混液對(duì)堿燒傷誘導(dǎo)的大鼠角膜新生血管的抑制作用及其作用機(jī)制研究[D];中國(guó)人民解放軍醫(yī)學(xué)院;2013年
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