CAJ | 학술논문

目的：体外建立高糖培养人视网膜色素上皮(hRPE)细胞模型,观察高糖条件下 RPE 细胞神经钙黏连蛋白(N-cadherin)和纤维连接蛋白( fibronectin)的表达变化,探讨高糖对RPE 细胞发生上皮间质转化(EMT)的影响。 　　方法：体外培养 hRPE 细胞,以60mmol / L 的葡萄糖建立高糖模型,实验分对照组(5.5mmol / L 的葡萄糖)和高糖24,48,72h 组,用免疫组织化学法及 Real-time PCR 技术检测各组 RPE 细胞 N-cadherin 和 fibronectin 的表达。 　　结果：高糖组 RPE 细胞出现排列紊乱,胞体变长变大,高糖72h 组最明显。免疫组织化学法检测结果显示：与对照组相比,高糖组 RPE 细胞 N-cadherin 表达呈下降趋势；而fibronectin 表达出现并呈升高趋势。 Real-time PCR 检测结果显示：与对照组相比,高糖组 RPE 细胞 N -cadherin mRNA 表达水平在24h 时出现下降,72h 时降低最明显(F =12.252,P=0.000),24h 组与对照组相比差异无统计学意义,48h 及72h 组分别与对照组相比差异有统计学意义(P<0.05)；与对照组相比,高糖组 RPE 细胞 fibronectin mRNA表达水平在24h 时出现升高,72h 时升高最明显( F =50.543,P=0.000),24h 组与对照组相比差异无统计学意义,48h 及72h 组 fibronectin mRNA 表达水平明显升高,分别与对照组相比差异有统计学意义(P=0.000,P=0.000)。结论：高糖条件下 RPE 细胞上皮标记物 N-cadherin 表达下调,间质标记物 fibronectin 表达出现,发生 EMT 改变,这一现象为探讨增生性糖尿病视网膜病变的发病机制及其防治提供新的思路。
목적：체외건립고당배양인시망막색소상피(hRPE)세포모형,관찰고당조건하 RPE 세포신경개점련단백(N-cadherin)화섬유련접단백( fibronectin)적표체변화,탐토고당대RPE 세포발생상피간질전화(EMT)적영향。 　　방법：체외배양 hRPE 세포,이60mmol / L 적포도당건립고당모형,실험분대조조(5.5mmol / L 적포도당)화고당24,48,72h 조,용면역조직화학법급 Real-time PCR 기술검측각조 RPE 세포 N-cadherin 화 fibronectin 적표체。 　　결과：고당조 RPE 세포출현배렬문란,포체변장변대,고당72h 조최명현。면역조직화학법검측결과현시：여대조조상비,고당조 RPE 세포 N-cadherin 표체정하강추세；이fibronectin 표체출현병정승고추세。 Real-time PCR 검측결과현시：여대조조상비,고당조 RPE 세포 N -cadherin mRNA 표체수평재24h 시출현하강,72h 시강저최명현(F =12.252,P=0.000),24h 조여대조조상비차이무통계학의의,48h 급72h 조분별여대조조상비차이유통계학의의(P<0.05)；여대조조상비,고당조 RPE 세포 fibronectin mRNA표체수평재24h 시출현승고,72h 시승고최명현( F =50.543,P=0.000),24h 조여대조조상비차이무통계학의의,48h 급72h 조 fibronectin mRNA 표체수평명현승고,분별여대조조상비차이유통계학의의(P=0.000,P=0.000)。결론：고당조건하 RPE 세포상피표기물 N-cadherin 표체하조,간질표기물 fibronectin 표체출현,발생 EMT 개변,저일현상위탐토증생성당뇨병시망막병변적발병궤제급기방치제공신적사로。
To observe the expression of N - cadherin and fibronectin in retinal pigment epithelium ( RPE) cells in vitro under high glucose conditions, furthermore, to explore the effects of high glucose on epithelial -mesenchymal transition (EMT) in RPE cells. ●METHODS: Human RPE (hRPE) cells were cultured in vitro. Containing a final concentration of 60mmol/ L glucose was used for high glucose treatment. The cells were divided into normal glucose group (5. 5mmol/ L, NG) and high glucose group (24, 48 and 72h) respectively. The expression of N - cadherin and fibronectin in hRPE cells were evaluated by immunofluorescence and real -time PCR. ●RESULTS:RPE cells became disorganized and swollen over time under high glucose conditions, especially in 72h subgroup. lmmunohistochemical analysis revealed that the expression of N - cadherin in RPE cells under high glucose conditions was decreased compared with that in the control group, while the expression of fibronectin was increased. Real - time PCR results showed that the expression of N - cadherin mRNA in high glucose group was decreased at 24h compared with that in the control group, and declined markedly at 72h ( F = 12. 252, P =0. 000). There were no significant differences between the control group and the high glucose group at 24h, while the differences between the control group and the high glucose group (48 and 72h) were significant respectively (P < 0. 05 ). Meanwhile, the expression of fibronectin mRNA in RPE cells was increased in high glucose group at 24h, and reached the peak at 72h (F = 50. 543, P = 0. 000). There were no significant differences between the control group and the high glucose group at 24h. Compared with the control group, the expression of fibronectin mRNA in hRPE cells was increased significantly in high glucose group (48 and 72h) respectively (P= 0. 000, P= 0. 000). ●CONCLUSlON: The expression of epithelium marker N-cadherin is down - regulated under high glucose conditions in hRPE cells in vitro. Meanwhile, the expression of mesenchymal maker fibronectin is induced and appeared to EMT changes. Results of this study will enrich our growing understanding in proliferative diabetic retinopathy and hopefully lead to novel insights for the pathogenesis and therapeutic treatments.