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- 학과정보 : 의학과 안과학전공 (검색결과 1,129 건)
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인체결막중층상피세포배양을 이용한 콕사키바이러스 결막염의 ex vivo 모델
Purpose: The purpose of this study was to establish an ex vivo model of coxsackievirus conjunctivitis since there seems to be no suitable disease model currently.. Methods: Human conjunctival epithelial cells (HCECs) were cultured for 2 weeks in a serum-free air-liquid interface system to produce a multi-layered structure. The cells were infected with coxsackievirus A24 (CVA24). Histological changes were investigated by staining the cells with H&E and DAPI, and apoptosis was evaluated using the TUNEL technique. Virus replication was measured in HeLa cells infected with viral progeny from multi-layered HCECs, after 1 and 3 days, using the TCID50 method. Results: Cultured HCECs formed multiple layers. The cells showed characteristics of conjunctival epithelial cells and goblet cells, being immunohistochemically positive for CK19 and MUC5AC, respectively. CVA24 replicated readily in cultured multi-layered HCECs. A mild cytopathic effect was noted 1 day after viral inoculation. Cell damage was extensive at 3 days. TUNEL imaging confirmed that the cytopathology was attributable to apoptosis. The TCID50 data from HeLa cells indicated that the virus was actively replicating at both the first and the third day after inoculation. Conclusions: The novel conjunctivitis model may be useful to investigate the pathogenesis of acute hemorrhagic conjunctivitis and the effectiveness of antiviral treatments.
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국문초록 만성 고안압 쥐 모델을 이용한 brinzolamide와 travoprost 점안 후 망막신경절세포 및 신경교세포의 변화 이 동 은 (안과학 전공) (지도교수 : 이주화) 의학과 인제대학교 대학원 목적: 만성 고안압 쥐 모델을 이용하여 brinzolamide (Azopt)와 travoprost (Travatan) 점안 후 발생하는 안압 하강 효과와 신경망막 변화에 대해 알아보고자 하였다. 방법: Adult male Sprague-Dawley rats 30마리( 녹내장군 15마리와 대조군 15마리) 60안을 대상으로 하였다. 각군을 각각 3개의 실험군으로 나누었다. 3개의 상공막 정맥 소작술로 만성적 고안압을 유발시켰다. Terminal deoxynucleotidyl transferase-mediated dUTP-nick end-labeling (TUNEL) 염색과 glial fibrillary acidic protein (GFAP) 발현법을 brinzolamide 와 travoprost 를 점안한 만성 고안압 쥐의 망막에 적용하였으며, bcl-2 family와 extracellular signal-regulated kinases (ERK)를 Western blot 으로 분석하여 망막변화의 내인성 신호전달 경로를 확인하였다. 결과: brinzolamide 와 travoprost 를 점안하여 고안압을 치료한 결과, 대조군에 비하여 의미 있는 안압 감소를 가져왔다. GFAP 발현법과 TUNEL 염색 상에서 brinzolamide 와 travoprost 를 점안하였을 때 망막신경절세포의 세포자멸사가 의미있게 감소하였으며, 신경교세포의 활성화가 이루어졌다. Bcl-2와 bcl-xL의 발현은 고안압을 유발시킨 후 의미있게 증가하였으며, brinzolamide 와 travoprost 점안 후에 더욱 증가하였다. 결론: 세포 생존 신호 전달의 강화는 망막신경절세포의 세포자멸사 감소에 기여할 수 있는 것으로 판단된다. 그러나 ERK신호전달의 역할은 뚜렷하지 않았다. 만성 고안압을 유발한 동물실험 모델에서 망막 손상의 감소는 brinzolamide 와 travoprost 의 신경보존적 치료 효과를 보여 주었으며, 임상적 이용을 위해서는 추후 더 많은 연구가 이루어져야 할 것으로 사료된다. 주제어 : 만성고안압, brinzolamide, neuroprotection, retinal glial fibrillary acidic protein expression, terminal deoxynucleotidyl transferase-mediated dUTP-nick end-labeling staining, travoprost Abstract Changes of the retinal ganglion cells and glial cells by brinzolamide and travoprost in chronic ocular hypertension rat model Dongeun Lee (Ophthalmology) (Advisor : Prof. Joo Hwa Lee, M.D.) Department of Medicine Graduate School, Inje University Objectives: The purpose of this study was to investigate the intraocular pressure-lowering effect and neuroretinal changes by brinzolamide (Azopt) and travoprost (Travatan) in a chronic ocular hypertension rat model. Methods: Thirty adult male Sprague-Dawley rats were used in this study (15 glaucoma group and 15 control group). For each experimental procedure, glaucoma and control groups consisted of 5 per group. Chronic ocular hypertension was induced by three episcleral vein cauterization. Terminal deoxynucleotidyl transferase-mediated dUTP-nick end-labeling (TUNEL) staining and expression of glial fibrillary acidic protein (GFAP) was performed in brinzolamide and travoprost treated chronic ocular hypertensive rat retina. Western blot analyses of bcl-2 family and extracellular signal-regulated kinases (ERK) were done to find out the intrinsic signaling pathway of the retinal changes. Results: Treatment of ocular hypertensive eyes with brinzolamide and travoprost significantly reduced intraocular pressure (25.8±1.4 mmHg and 23.6±1.7 mmHg) compared to control eyes (31.1±1.7 mmHg). GFAP expression and TUNEL stain revealed significant decreases in retinal ganglion cell (RGC) apoptosis and glial cell activation after brinzolamide and travoprost administration. bcl-2 and bcl-xL expression were significantly increased after intraocular pressure elevation and it was further increased with brinzolamide and travoprost treatment. Conclusions: The enhancement of cell survival signaling might have contributed to the decrease of RGCs apoptosis. However, the role of ERK signaling was not evident in this process. Decrease in retinal damage suggests the neuroprotective potential of brinzolamide and travoprost in the animal model of chronic ocular hypertension, but further studies are required before clinical use of these agents. Key Words : Brinzolamide, chronic ocular hypertension, neuroprotection, retinal glial fibrillary acidic protein expression, terminal deoxynucleotidyl transferase-mediated dUTP-nick end-labeling staining, travoprost
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(초 록) 약시가 망막 두께에 영향을 주는지 알아보기 위하여 정상 소아와 약시 소아의 중심와 및 망막신경섬유층 두께를 측정하였다. 굴절부등, 사시로 인한 한눈 약시 소아 26명(52안)과 정상 소아 42명(84안), 총 136안을 대상으로 빛간섭단층촬영기(Optical Coherence Tomography, Version 3.0, USA)를 시행하였다. 망막 두께의 측정을 중심와에서, 망막신경섬유층은 시신경유두 주위의 상측, 하측, 비측, 이측에서 시행하였다. 연령은 정상 소아의 경우 평균 8.5세, 약시 소아는 평균 8.0세였다. 중심와 두께는 정상 소아와 약시안의 두 군 간에 차이가 없었다. 망막신경섬유층 두께는 상측, 하측, 비측, 이측에서 정상 소아와 약시 소아 간의 차이는 없었다. 그러나 굴절부등약시와 사시약시 소아간의 중심와와 망막신경섬유 두께는 차이가 있었으나 약시가 중심와 및 전체적인 망막신경섬유층 두께에는 영향을 주지 않았다. (Abstract) We measured the thickness of the fovea and the retinal nerve fiber layer of normal children and the children with amblyopia to find whether amblyopia have an effect on the retinal thickness. Optical coherence tomography (OCT, Version 3.0, USA) was performed on 26 children (52 eyes) with unilateral amblyopia that was due to anisometropia or strabismus, and it was also performed on 42 normal children (84 eyes), for a total of 136 eyes. Retinal thickness measurement was performed on the fovea, and the retinal nerve fiber layer thickness measurement was performed on the superior, inferior, nasal and temporal quadrants in the peripapillary region. The age of the normal children was an average of 8.5 years and the age of the children with amblyopia was an average of 8.0 years. The difference of the thickness of fovea between the normal children and amblyopic eye was not statistically significant. The difference of thickness of the superior, inferior, nasal and temporal quadrants of the retinal nerve fiber layer between the normal children and the children with amblyopia was not statistically significant. However, the thickness of the fovea and the retinal nerve fiber layer were significant difference between the children with anisometropic amblyopia and the children with strabismic amblyopia. But, it was found that amblyopia does not have an effect on the thickness of the fovea and the total retinal nerve fiber layer.
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역기하렌즈 ; 근시교정효과 및 각막상피세포와 기질에 미치는 영향
Orthokeratology is the technique of fitting reverse geometry lenses in order to change the corneal curvature as a way of improving myopia. This study is designed to evaluate the level of myopia correction and safety of Ortho-K LK^(TM) lenses and the effect of reverse geometry lens to corneal epithelium and stroma. A through and retrospective analysis was performed on 17 patients (34 eyes), who were wearing Ortho-K LK^(TM) lenses. In order to track the changes in the level of uncorrected visual acuity, spherical equivalent, astigmatism, and corneal endothelial cell count, we performed a periodical analysis, before the fitting and after the fitting, based on 1 week, 2 week, 1 month, 3 month and 6 month-term. The reverse geometry lenses were fitted to the right eyes of 15 rabbits and RGP lenses to the other 15 rabbits to compare the effects of reverse geometry lenses to cornea epithelium and stroma. Rabbits were sacrificed on the day of the first, third, seventh, tenth and fourteenth days after lens fitting. The thickness of the cornea was measured by microscope and microscale and the ratio of right and left corneal thickness was also considered in order to eliminate the deviation of difference in the individual cornea. In addition, to prove the migration and redistribution of corneal epithelial cells, we observed the filamentous actin in corneal epithelium after immunofluorescent stain, using NBD phallacidin. A significant progress developed that the uncorrected visual acuity and spherical equivalent improved in a week and it persisted in 6 months. But, no significant changes were found in the amount of astigmatism and corneal endothelial cell density. However, the visual acuity had been decreased in 3 to 6 months after the fitting, therefore we assumed that there must be another reason beyond the patients' compliance. The most common examples among the known complications were the decenteration of lenses, corneal erosion and corneal iron deposit. However, there were no such serious complications as to stop lens weanng. Furthermore, no consistent changes in the thickness of corneal epithelium and stroma at center and periphery were found, which can be explained by the severe corneal edema due to the persistent lens fitting and decenteration of lenses in the cornea of the rabbits. We did observe the positive NBD phallacidin immunofluorescent stain in both of the groups with the reverse geometry lenses and RGP lenses. But, on the cornea with the reverse geometry lenses showed a stronger reaction. Thus, we concluded that the migration of corneal epithelium under the effect of reverse geometry lenses changed the anterior corneal curvature.
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