本文選題：脂肪干細(xì)胞 + 青光眼　； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的:研究脂肪干細(xì)胞玻璃體腔注射對(duì)大鼠青光眼模型的視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的保護(hù)作用,并初步探討可能的機(jī)制。方法:培養(yǎng)脂肪干細(xì)胞,培養(yǎng)至P4接近單層融合時(shí),收集并調(diào)整細(xì)胞密度備用。將大鼠分為四組:青光眼干細(xì)胞組、青光眼對(duì)照組、正常眼壓干細(xì)胞組和正常眼壓對(duì)照組,采用烙閉表層鞏膜靜脈的方法制備大鼠青光眼模型,均以右眼為操作眼,青光眼干細(xì)胞組、青光眼對(duì)照組行造模手術(shù),正常眼壓干細(xì)胞組、正常眼壓對(duì)照組行假手術(shù)操作。手術(shù)操作3周時(shí),三組均行玻璃體腔注射操作,青光眼干細(xì)胞組注射物為5×105 ADSCs懸浮液6?L,青光眼對(duì)照組注射物為DMEM液6?L,正常眼壓對(duì)照組注射物為5×105 ADSCs懸浮液6?L,正常眼壓對(duì)照組注射物DMEM液6?L。分別測(cè)量并記錄術(shù)前、術(shù)后1周、2周、3周、4周、5周、6周眼壓。實(shí)驗(yàn)完成后將動(dòng)物處死,隨機(jī)摘取眼球,分別進(jìn)行冰凍切片HE染色、免疫熒光染色處理,并進(jìn)行q RT-PCR檢測(cè)BDNF m RNA表達(dá)量。結(jié)果:造模后1周,模型眼相對(duì)于假手術(shù)眼,IOP平均升高10.9±2.1mm Hg,并能在預(yù)定實(shí)驗(yàn)時(shí)間6周內(nèi)保持這一水平。而在對(duì)各組進(jìn)行移植操作后,青光眼干細(xì)胞組的IOP降低水平在移植操作后1、2、3周時(shí)均顯著高于青光眼對(duì)照組(p0.001)。HE染色青光眼干細(xì)胞組、青光眼對(duì)照組與正常眼壓干細(xì)胞組、正常眼壓對(duì)照組相較均出現(xiàn)后極部視網(wǎng)膜各層變薄,RGC層細(xì)胞排列紊亂,RGC數(shù)量減少,但青光眼對(duì)照組較青光眼干細(xì)胞組病理改變嚴(yán)重。免疫熒光染色兩組青光眼組與兩組正常眼壓組相較,熒光染色淡,神經(jīng)節(jié)細(xì)胞數(shù)明顯減少,青光眼對(duì)照組較青光眼干細(xì)胞組減少更明顯。除正常眼壓兩組間外,其他各組間在400倍熒光顯微鏡下RGC計(jì)數(shù)差異均具有統(tǒng)計(jì)學(xué)意義,計(jì)數(shù):青光眼對(duì)照組青光眼干細(xì)胞組正常眼壓干細(xì)胞組/正常眼壓對(duì)照組。BDNF m RNA各組之間差異具有統(tǒng)計(jì)學(xué)意義(*P0.05;**P0.01;***P0.001),兩兩比較差異具有統(tǒng)計(jì)學(xué)意義,且BDNF m RNA表達(dá)量:青光眼干細(xì)胞組正常眼壓干細(xì)胞組青光眼對(duì)照組正常眼壓對(duì)照組。結(jié)論:脂肪干細(xì)胞玻璃體腔移植對(duì)大鼠青光眼模型視網(wǎng)膜神經(jīng)節(jié)細(xì)胞具有保護(hù)作用,可能通過降低眼壓和增多BDNF等途徑發(fā)揮保護(hù)作用。
[Abstract]:Aim: to study the protective effect of adipose stem cells intravitreal injection on retinal ganglion cells (RGCs) in rat glaucoma model and to explore the possible mechanism. Methods: adipose stem cells were cultured until P 4 was close to monolayer fusion. Cell density was collected and adjusted. Rats were divided into four groups: glaucoma stem cell group, glaucoma control group, normal intraocular pressure stem cell group and normal intraocular pressure control group. Glaucoma stem cell group, glaucoma control group, normal IOP stem cell group, normal IOP control group were performed sham operation. After 3 weeks of operation, all the three groups were treated with intravitreal injection. In the glaucoma stem cell group, 5 脳 10 5 ADSCs suspension was 6L, the glaucoma control group was DMEM 6L, the normal intraocular pressure control group was 5 脳 105 ADSCs suspension 6L, and the normal intraocular pressure control group was DMEM 6L. Intraocular pressure was measured and recorded 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks and 6 weeks after operation. After the experiment, the animals were killed, eyeballs were randomly removed, frozen sections were stained with HE and immunofluorescence staining, and the expression of BDNF mRNA was detected by qRT-PCR. Results: the IOP of the model eyes increased by 10.9 鹵2.1mm Hg 1 week after the model compared with the sham operation eyes, and could be maintained within 6 weeks of the scheduled experimental time. After transplantation, IOP levels in glaucoma stem cell group were significantly higher than those in glaucoma control group (p0.001), glaucoma control group and normal IOP stem cell group. Compared with the normal intraocular pressure control group, the number of RGC cells in the RGC layer of the posterior polar retinal thinning was decreased, but the pathological changes in the glaucoma control group were more serious than those in the glaucoma stem cell group. Compared with normal intraocular pressure group, immunofluorescence staining showed that the number of ganglion cells in glaucoma group was less than that in normal intraocular pressure group, and the number of ganglion cells in glaucoma control group was significantly lower than that in glaucoma stem cell group. With the exception of normal intraocular pressure (IOP), there were significant differences in RGC counts between the other groups under a 400-fold fluorescence microscope. Counting: the difference of BDNF mRNA between normal IOP stem cell group and normal IOP control group was statistically significant (P0.05, P0.01, P0.001), and the difference between two groups was statistically significant. The expression of BDNF mRNA: normal IOP stem cell group, glaucoma control group, normal IOP control group. Conclusion: vitreous transplantation of adipose stem cells has protective effect on retinal ganglion cells in rat model of glaucoma, which may play a protective role by reducing intraocular pressure and increasing BDNF.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R775;R-332

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相關(guān)期刊論文 前3條

1 劉子琪;孫同文;萬有棟;姚瑞;闞全程;;脂肪源性干細(xì)胞體外分離培養(yǎng)及向內(nèi)皮祖細(xì)胞的誘導(dǎo)分化[J];中國組織工程研究;2015年32期

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本文編號(hào)：2059995