【摘要】： 第一部分皮下注射氫溴酸東莨菪堿建立兔干眼模型的實(shí)驗(yàn)研究 目的:探討兔皮下注射氫溴酸東莨菪堿,建立干眼模型的特點(diǎn)和眼表病理?yè)p害機(jī)制。 方法:健康新西蘭大白兔18只,隨機(jī)分為三組,每組6只。實(shí)驗(yàn)組皮下注射氫溴酸東莨菪堿,其中低劑量組(L,1.0mg/次)、高劑量組(H, 2.0mg/次),對(duì)照組注射等量生理鹽水。注射時(shí)間點(diǎn)為每天8點(diǎn)11點(diǎn)、14點(diǎn)和18點(diǎn),連續(xù)30天。用藥后0、7,14,21,30d對(duì)實(shí)驗(yàn)組及對(duì)照組行淚液分泌試驗(yàn)、淚膜破裂時(shí)間(BUT)、角膜熒光素染色,印跡細(xì)胞檢查。第30d空氣栓塞法處死實(shí)驗(yàn)動(dòng)物,取角膜、結(jié)膜及淚腺組織行HE染色,免疫組化檢測(cè)FAS,BCL-2及NFκB)的表達(dá)水平。 結(jié)果:酚紅棉線法測(cè)得高劑量(H)組造模前淚液分泌量為16.25±2.299mm,第21d時(shí)下降為6.75±1.982mm,對(duì)照組為16.50±2.619mm,差異具有統(tǒng)計(jì)學(xué)意義(p0.05),同期低劑量組(L)為15.00±2.390mm,與對(duì)照組相比無(wú)明顯差異。H組第3d出現(xiàn)BUT縮短,L組則第7d發(fā)生BUT的變化,此后BUT值均低于5s,對(duì)照組BUT大于20s。實(shí)驗(yàn)組均出現(xiàn)角膜熒光素染色陽(yáng)性,H組出現(xiàn)時(shí)間早(3d),持續(xù)時(shí)間長(zhǎng)。L組和H組PAS染色與對(duì)照組相比,杯狀細(xì)胞形態(tài)紊亂、核淡染,部分核固縮,L組和H組之間無(wú)明顯差異。HE染色顯示H組淚腺周?chē)?jiàn)散在淋巴細(xì)胞和腺腔凋亡性改變,角膜上皮呈重度瘤樣增生基質(zhì)層變性伴水腫,結(jié)膜杯狀細(xì)胞反應(yīng)性增多,固有層見(jiàn)淋巴細(xì)胞浸潤(rùn)數(shù)量增加。L組角膜淺層上皮輕度增生,細(xì)胞排列稍紊亂,結(jié)膜和淚腺組織病理改變輕微。電鏡觀察高劑量組淚腺腺管管腔壁絨毛較對(duì)照組明顯減少,角膜表層微絨毛較對(duì)照組減少,上皮細(xì)胞空泡變,細(xì)胞核固縮,結(jié)膜表面微絨毛減少,上皮細(xì)胞呈壞死凋亡改變,低劑量未見(jiàn)明顯顯微改變。凋亡抑制因子BCL-2在高劑量模型組淚腺、角膜和結(jié)膜組織中極少表達(dá),對(duì)照組各組織均呈高度陽(yáng)性表達(dá),兩組差異顯著p0.001;促凋亡因子Fas在模型組淚腺組織中呈中度表達(dá),在角膜和結(jié)膜組織中呈中高度表達(dá)。炎癥轉(zhuǎn)錄因子NFκB在模型組兔眼淚腺腺泡細(xì)胞胞漿和胞核輕度表達(dá),腺腔管壁細(xì)胞胞核和胞漿中重度表達(dá),角膜組織上皮細(xì)胞和基底細(xì)胞胞漿、胞核以及結(jié)膜上皮細(xì)胞中NFκB呈中度陽(yáng)性表達(dá),對(duì)照組各組織NFκB極少表達(dá),兩組差異有統(tǒng)計(jì)學(xué)意義(p0.001)。 結(jié)論:兔皮下注射氫溴酸東莨菪堿,4次/天,2.0mg/次,可有效維持藥物外周濃度,抑制淚腺的分泌,引起角結(jié)膜上皮細(xì)胞的損傷以及角結(jié)膜上皮和淚腺組織細(xì)胞凋亡因子和炎癥因子陽(yáng)性表達(dá)增加,成功建立以淚液缺乏為主,凋亡和炎癥反應(yīng)共同參與的干眼動(dòng)物模型。 第二部分FK506對(duì)氫溴酸東莨菪堿至兔干眼模型的療效 目的:觀察FK506滴眼液干預(yù)兔干眼模型的療效及其作用機(jī)制 方法:健康新西蘭大白兔30只,,隨即分為實(shí)驗(yàn)組18只和對(duì)照組12只。實(shí)驗(yàn)組采用上述方法每天8點(diǎn)11點(diǎn)、14點(diǎn)和18點(diǎn)皮下注射氫溴酸東莨菪堿注射液2.0mg/次,并從第四天開(kāi)始左眼點(diǎn)0.05%FK506滴眼液,4次/天,右眼不做處理,對(duì)照組皮下注射生理鹽水。實(shí)驗(yàn)第0,3,7,14,21,35d對(duì)實(shí)驗(yàn)組及對(duì)照組行淚液分泌試驗(yàn)、淚膜破裂時(shí)間(BUT)、角膜熒光素染色檢查。分別在第7,14,和35d采取空氣栓塞法分別隨即處死模型組5只兔子,取雙眼角膜、結(jié)膜和淚腺組織,PCR定量分析IL-1β、TNF-α和MMP-9等炎性因子在各組織的表達(dá)。第35d采用印記細(xì)胞學(xué)方法取雙眼角結(jié)膜上皮細(xì)胞行免疫組化檢測(cè)NFκB表達(dá)水平的差異。 結(jié)果:造模后3,7天實(shí)驗(yàn)兔雙側(cè)眼均角膜熒光素染色強(qiáng)陽(yáng)性。第14d開(kāi)始FK506治療眼角膜熒光素染色評(píng)分低于未用藥眼,至第35d,差異進(jìn)一步增大具有統(tǒng)計(jì)學(xué)意義(P0.01)。FK506治療組眼的角結(jié)膜上皮NFκB表達(dá)顯著低于對(duì)照眼,不同時(shí)間點(diǎn)治療組結(jié)膜組織中IL-1β、TNF-α和MMP-9表達(dá)水平低于對(duì)照組,兩者差異有統(tǒng)計(jì)學(xué)意義(p0.05)。14天時(shí)雙側(cè)淚腺M(fèi)MP-9水平造模后與對(duì)照組相比呈現(xiàn)增高,但用藥干預(yù)眼與未治療眼之間無(wú)明顯差異。 結(jié)論:FK506通過(guò)抑制炎癥信號(hào)分子NFκB的激活,降低角膜,結(jié)膜組織中IL-1β、TNF-α和MMP-9等炎癥因子的水平,有效地抑制眼表免疫性炎癥,進(jìn)而減輕干眼對(duì)眼表組織的損害。
[Abstract]:The experimental study on the establishment of rabbit model of dry eye by subcutaneous injection of the first part by subcutaneous injection of hydrobromic acid Objective: To study the characteristics of the dry eye model and the pathology of ocular surface in rabbits by subcutaneous injection of hydrobromic acid. Damage mechanism. Methods: 18 healthy New Zealand white rabbits, randomized The experimental group was divided into three groups, each group of 6 rats. The experimental group was injected with hydrobromic acid, the lower dose group (L, 1. 0mg/ times) and the high dose group (H, 2. 0mg/ time), and the control group. The same amount of saline was injected into the group. The injection time point was 8: 11, 14, and In the experimental group and the control group, the tear secretion test, the tear film rupture time (BUT) and the corneal fluorescein were measured at 18: 0, 7, 14, 21, and 30 days after the administration. The cells were stained and blotted. The experimental animals were sacrificed by the method of air embolism on the 30th day. The cells of the cornea, conjunctiva and lacrimal gland were stained with HE, and the FAS, BCL-2 and N were detected by immunohistochemistry. Results: The amount of tear secretion in the high-dose (H) group was 16.25-2.299mm in the high-dose (H) group and 6.75-1.982mm in the control group and 16.50-2.619mm in the control group (P 0.05), while the low-dose group (L) was 15.00-2.390 in the same period. There was no significant difference between the control group and the control group. In the H group, the BUT was shortened in the third day, and the changes of the BUT occurred in the group L, and the BUT value was lower after that. In the control group, the BUT of the control group was more than 20s. In the experimental group, the staining of the corneal fluorescein was positive, and the H group showed a positive staining. The present time was early (3d) and the duration was long. The PAS staining of the L and H groups was compared with that of the control group. There was no significant difference between the group L and the H group. HE staining showed that there was a change in the apoptosis of the lymphocytes and the gland cavity around the lacrimal gland of the H group, the changes of the stromal layer with severe tumor-like hyperplasia and edema in the corneal epithelium, and the increase of the reactivity of the conjunctiva cup-like cells. Inherent layer was found to increase in the number of infiltration of lymphocytes. The superficial epithelium of the L group was slightly hyperplastic and the cells were arranged slightly. The pathological changes of the conjunctiva and the lacrimal gland were mild. The microvilli in the lumen of the lacrimal gland of the high-dose group were significantly reduced in the high-dose group, the microvilli in the surface of the cornea decreased, the vacuolation of the epithelial cells was reduced, the cell nucleus was fixed, the microvilli on the surface of the conjunctiva were reduced, and the epithelial cells were necrotic. The apoptosis-inhibiting factor BCL-2 was expressed in the lacrimal gland, the cornea and the conjunctival tissue of the high-dose model group, and the control group was highly positive, and the difference between the two groups was significantly higher than that of the control group, and the difference between the two groups was significantly higher than that of the control group. The expression of the inflammatory transcription factor NF-B in the cytoplasm and the nucleus of the cell and the cytoplasm of the cell and the cytoplasm of the cell, the cytoplasm, the nucleus and the conjunctival epithelium of the epithelial cell and the basal cell of the rabbit's lacrimal gland in the model group were observed in the model group. NF-B in the control group was expressed moderately, and NF-B in the control group was very rarely expressed, and the difference between the two groups Conclusion: The peripheral concentration of the drug can be effectively maintained, the secretion of the lacrimal gland, the damage to the conjunctival epithelial cells and the conjunctival epithelium and the lacrimal gland can be effectively maintained. The positive expression of the cell apoptosis factor and the inflammatory factor increased, and the success was based on the lack of tear. A dry-eye animal model that is co-involved in the reaction of apoptosis and inflammation. The effect of two-part of FK506 on the dry-eye model in the eastern and eastern part of the rabbit Objective: To observe the effect and mechanism of FK506 eye drops in the dry eye model of rabbits Methods: Thirty-one healthy New Zealand white rabbits were divided into experimental group and control group. 5% FK506 eye drops, 4 times/ day, right eye not treated, control group subcutaneous injection of physiological saline. Experiment Days 0, 3, 7, 14, 21, 35d In the group and control group, the tear secretion test, the tear film rupture time (BUT) and the corneal fluorescein staining were performed. 5 rabbits were respectively sacrificed in the model group at the 7th, 14th, and 35d respectively, and the two eyes of the cornea, the conjunctiva and the lacrimal gland tissue and the PC were taken. R. Quantitative analysis of the expression of IL-1, TNF-1 and MMP-9 in the tissues. A Cytological Approach to the Expression of NF-B in the Conjunctival Epithelium of the Binocular Conjunctival Epithelial Cells The results showed that in 3 and 7 days after the model, the two-side eyes of the experimental rabbits were stained with fluorescein. At the same time, the staining score of the eye-cornea fluorescein was lower than that of the non-treated eyes at the beginning of the 14th day, and the difference between the two groups was statistically significant (P0.01). The FK506 treatment group The expression of IL-1, TNF-1 and MMP-9 in the conjunctival tissue of the treatment group was lower than that of the control group. Conclusion: FK506 can reduce the level of IL-1, TNF and TNF in the cornea and conjunctival tissue by inhibiting the activation of NF-B in the inflammatory signal.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R777

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 黃秀英,楊紅,龔桂芝,王林娟,郭曉霞;FK506抗同種腎移植排斥反應(yīng)的肝功能觀察及護(hù)理[J];護(hù)士進(jìn)修雜志;2002年04期

2 陳國(guó)奮,顧立強(qiáng);FK506加速周?chē)窠?jīng)損傷修復(fù)后的功能恢復(fù)[J];中華手外科雜志;2002年02期

3 夏正坤 ,劉光陵 ,高遠(yuǎn)賦 ,樊忠民 ,伏潔 ,傅元鳳 ,張連豐;FK506治療兒童不同病理類(lèi)型腎病綜合征臨床觀察[J];臨床兒科雜志;2003年10期

4 王志東,顧軍,米慶勝;他克莫司對(duì)TNF-α誘導(dǎo)的HaCaT細(xì)胞NO分泌的影響[J];中國(guó)麻風(fēng)皮膚病雜志;2002年03期

5 趙京霞,王為忠,宋維亮,吳國(guó)生,羅寧寧,陳新莉,徐菊麗;小腸移植術(shù)后腸內(nèi)營(yíng)養(yǎng)因素對(duì)FK506血藥濃度的影響[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2000年06期

6 于立新,劉小友,胡敏燕;FK506的藥代動(dòng)力學(xué)及臨床研究[J];第一軍醫(yī)大學(xué)學(xué)報(bào);2000年06期

7 陳浩宇,510060?,楊培增,510060?;FK506的免疫抑制作用及其在葡萄膜炎治療中的應(yīng)用[J];眼科研究;2000年05期

8 祖強(qiáng),姚冰,于立新;FK506的腎毒性[J];腎臟病與透析腎移植雜志;2000年03期

9 丁小明,薛武軍,田普訓(xùn),項(xiàng)和立;FK506治療乙肝病毒感染的腎移植受者排斥反應(yīng)的臨床觀察[J];腎臟病與透析腎移植雜志;2002年04期

10 張衛(wèi)國(guó),傅重洋;FK506對(duì)大鼠坐骨神經(jīng)再生作用的實(shí)驗(yàn)研究[J];實(shí)用手外科雜志;2004年01期

相關(guān)會(huì)議論文 前10條

1 孫化萍;于曉林;曾慶華;羅旭升;楊紅;;黃精多糖滴眼液對(duì)干眼模型結(jié)膜的影響[A];第三次全國(guó)中醫(yī)、中西醫(yī)結(jié)合眼科學(xué)術(shù)交流會(huì)論文匯編[C];2003年

2 白小良;曾濤;謝淑英;;LASIK術(shù)后鼻側(cè)蒂與上方蒂引起干眼的比較[A];中華醫(yī)學(xué)會(huì)第十二屆全國(guó)眼科學(xué)術(shù)大會(huì)論文匯編[C];2007年

3 劉祖國(guó);邱璇;梁凌毅;劉揚(yáng);;干眼患者焦慮、抑郁心理調(diào)查[A];中華醫(yī)學(xué)會(huì)第十二屆全國(guó)眼科學(xué)術(shù)大會(huì)論文匯編[C];2007年

4 陳蔚;陳金鵬;張曉博;王世明;張京娜;瞿佳;;TLR9在干眼綜合癥患者結(jié)膜上皮細(xì)胞中的表達(dá)[A];中華醫(yī)學(xué)會(huì)第十二屆全國(guó)眼科學(xué)術(shù)大會(huì)論文匯編[C];2007年

5 曾慶延;王江橋;張捷;秦光勇;牛曉光;喻長(zhǎng)泰;;偉博水霧劑治療干眼的臨床觀察[A];2005年浙江省眼科學(xué)術(shù)會(huì)議論文集[C];2005年

6 陳金鵬;陳蔚;張曉博;王世明;張京娜;瞿佳;;HLA-DR在不同程度干眼綜合癥患者結(jié)膜上皮細(xì)胞中的表達(dá)[A];中華醫(yī)學(xué)會(huì)第十二屆全國(guó)眼科學(xué)術(shù)大會(huì)論文匯編[C];2007年

7 陳小平;劉祖國(guó);梁凌毅;;Sjgren綜合征LASIK術(shù)后嚴(yán)重干眼6例臨床分析[A];中華醫(yī)學(xué)會(huì)第十二屆全國(guó)眼科學(xué)術(shù)大會(huì)論文匯編[C];2007年

8 吳平;劉建;張忠濤;張棟;王宇;;中低血藥濃度下Fk506在肝移植中的應(yīng)用[A];第一屆全國(guó)人工肝及血液凈化學(xué)術(shù)年會(huì)暨全國(guó)人工肝及血液凈化攻關(guān)協(xié)作組成立大會(huì)論文集[C];2004年

9 王貴平;趙忠新;莊建華;黃流清;吳惠涓;張琳;彭華;由振東;;REM睡眠剝奪對(duì)大鼠空間記憶、海馬CaN的影響及FK506的干預(yù)作用[A];第三屆中國(guó)睡眠醫(yī)學(xué)論壇論文匯編[C];2009年

10 楊光;元小玲;;淚河寬度定量檢查與干眼相關(guān)性分析[A];第六屆全國(guó)中醫(yī)中西醫(yī)結(jié)合眼科學(xué)術(shù)交流會(huì)論文匯編[C];2007年

相關(guān)重要報(bào)紙文章 前10條

1 董晨　顧德寧;當(dāng)心電腦帶來(lái)“集體干眼”癥[N];新華日?qǐng)?bào);2004年

2 中國(guó)中醫(yī)科學(xué)院眼科醫(yī)院 楊曉樺;中醫(yī)藥內(nèi)服外熏蒸治療干眼[N];中國(guó)中醫(yī)藥報(bào);2008年

3 張素蘭;態(tài)度決定高度[N];天津日?qǐng)?bào);2004年

4 朱媛媛;口干眼燥應(yīng)及時(shí)就診[N];醫(yī)藥養(yǎng)生保健報(bào);2006年

5 ;燒菜小心“醉油”[N];中華合作時(shí)報(bào);2003年

6 張曼凌;用信任的目光注視孩子[N];中國(guó)教師報(bào);2003年

7 東方;胡蘿卜下酒 毒！[N];保健時(shí)報(bào);2003年

8 謝幼紅 陳愛(ài)萍;口干眼干別大意[N];健康報(bào);2006年

9 珞珈投資 馮明波;2003：哪些板塊會(huì)有戲？[N];中國(guó)石化報(bào);2003年

10 顧德寧;不孕不是錯(cuò)，，更不是“罪”[N];新華日?qǐng)?bào);2005年

相關(guān)博士學(xué)位論文 前10條

1 盧安衛(wèi);以FK506為基礎(chǔ)的免疫抑制方案在肝移植后應(yīng)用的合理性研究[D];浙江大學(xué);2005年

2 陳國(guó)奮;FK506加速大鼠坐骨神經(jīng)橫斷傷后功能恢復(fù)及其機(jī)制的實(shí)驗(yàn)研究[D];第一軍醫(yī)大學(xué);2007年

3 孫佳冰;異種周?chē)窠?jīng)移植免疫耐受的實(shí)驗(yàn)研究[D];吉林大學(xué);2009年

4 楊海云;脊髓骨髓間充質(zhì)干細(xì)胞促進(jìn)損傷脊髓修復(fù)和神經(jīng)干細(xì)胞的分化[D];吉林大學(xué);2009年

5 李明新;FK506及神經(jīng)干細(xì)胞對(duì)異體神經(jīng)移植作用的研究[D];天津醫(yī)科大學(xué);2009年

6 闕軍;FK506通過(guò)減少瘢痕形成促進(jìn)周?chē)窠?jīng)再生的作用及機(jī)制研究[D];南京醫(yī)科大學(xué);2012年

7 李洪敬;川芎嗪、FK506對(duì)犬急性脊髓損傷神經(jīng)保護(hù)作用的實(shí)驗(yàn)研究[D];大連醫(yī)科大學(xué);2005年

8 肖鶴;用于小分子化合物篩選的細(xì)胞模型的建立與鑒定[D];中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院;2005年

9 邢宏義;腦缺血后神經(jīng)可塑性與修復(fù)的研究[D];華中科技大學(xué);2006年

10 馮星;心肌缺血/再灌注損傷及鈣調(diào)神經(jīng)磷酸酶信號(hào)通路的調(diào)控作用[D];吉林大學(xué);2009年

相關(guān)碩士學(xué)位論文 前10條

1 仇晶晶;兔干眼模型的建立及FK506治療干眼的療效及作用機(jī)制研究[D];南昌大學(xué);2010年

2 張賀;干眼Ⅰ號(hào)方治療肝腎陰虛型干眼的療效觀察[D];北京中醫(yī)藥大學(xué);2010年

3 董學(xué)梅;中藥治療白內(nèi)障超聲乳化術(shù)后干眼的臨床觀察[D];南京中醫(yī)藥大學(xué);2012年

4 陳妍遐;轉(zhuǎn)化生長(zhǎng)因子β1在干眼患者眼表的表達(dá)研究[D];山西醫(yī)科大學(xué);2012年

5 官宇;大連眼科門(mén)診眼干澀患者調(diào)查[D];大連醫(yī)科大學(xué);2011年

6 鄧妮妮;蒸發(fā)過(guò)強(qiáng)型干眼的抗炎對(duì)比研究[D];廣西醫(yī)科大學(xué);2010年

7 李影;眼周針刺治療水液缺乏性干眼的臨床研究[D];南京中醫(yī)藥大學(xué);2010年

8 張亞;海藻糖對(duì)大鼠蒸發(fā)過(guò)強(qiáng)型干眼作用機(jī)制及療效的實(shí)驗(yàn)研究[D];遼寧醫(yī)學(xué)院;2011年

9 柯于海;不同時(shí)段應(yīng)用FK506納米微球緩釋顆粒促進(jìn)同種異體神經(jīng)移植再生的實(shí)驗(yàn)研究[D];廣州醫(yī)學(xué)院;2010年

10 常靜;干眼患者結(jié)膜IL-1β的表達(dá)和意義[D];山西醫(yī)科大學(xué);2012年

本文編號(hào)：2314022