中华眼视光学与视觉科学杂志
中華眼視光學與視覺科學雜誌
중화안시광학여시각과학잡지
CHINESE JOURNAL OF OPTOMETRY OPHTHALMOLOGY AND VISUAL SCIENCE
2012年
7期
429-433
,共5页
近视,退行性%转甲状腺素蛋白%视网膜裂孔,黄斑%视网膜脱离,黄斑
近視,退行性%轉甲狀腺素蛋白%視網膜裂孔,黃斑%視網膜脫離,黃斑
근시,퇴행성%전갑상선소단백%시망막렬공,황반%시망막탈리,황반
Myopia,degenerative%Transthyretin%Retinal holes,macular%Retinal detachment,macular
目的 探讨转甲状腺素蛋白(TTR)在病理性近视患者玻璃体中表达及其结构分析.方法 实验研究.收集黄斑区脱离(n=14)和病理性近视黄斑裂孔(n=9)的玻璃体样本,另选正常角膜供体眼玻璃体样本(n=8)作为正常对照.对3组玻璃体进行了酶联免疫吸附试验(ELISA)、聚丙烯酰胺凝胶电泳(SDS-PAGE)、免疫印迹分析(western blot,WB)和圆二色谱(CD)分析.对所得数据进行单因素方差分析.结果 正常人、黄斑区脱离患者、黄斑裂孔患者玻璃体中TTR的浓度分别为(45.2±1.0)mg/L、(230.0±16.3)mg/L、( 109.8±19.0)mg/L,差异有统计学意义(F=2.69,P＜0.05).SDS-PAGE结果显示,黄斑区脱离患者玻璃体中TTR四聚体结构明显比黄斑裂孔患者和正常对照的稳定;对电泳条带进行黑度积分分析,黄斑区脱离患者玻璃体中TTR含量约为黄斑裂孔患者玻璃体TTR含量的2.4倍,与ELISA的结果相一致.CD结果显示黄斑区脱离和黄斑裂孔患者玻璃体TTR的二级结构分别含有45％α螺旋和32％α螺旋,且在不同pH(4～11)下保持稳定.正常样本玻璃体TTR的二级结构含有5％α螺旋,且随pH(4～11)变化产生巨大波动.结论 黄斑区脱离患者以及黄斑裂孔患者玻璃体中TTR含量较高,二级结构相对稳定,且黄斑区脱离患者玻璃体TTR四聚体结构牢固,这可能与蛋白变异和错误折叠有关.
目的 探討轉甲狀腺素蛋白(TTR)在病理性近視患者玻璃體中錶達及其結構分析.方法 實驗研究.收集黃斑區脫離(n=14)和病理性近視黃斑裂孔(n=9)的玻璃體樣本,另選正常角膜供體眼玻璃體樣本(n=8)作為正常對照.對3組玻璃體進行瞭酶聯免疫吸附試驗(ELISA)、聚丙烯酰胺凝膠電泳(SDS-PAGE)、免疫印跡分析(western blot,WB)和圓二色譜(CD)分析.對所得數據進行單因素方差分析.結果 正常人、黃斑區脫離患者、黃斑裂孔患者玻璃體中TTR的濃度分彆為(45.2±1.0)mg/L、(230.0±16.3)mg/L、( 109.8±19.0)mg/L,差異有統計學意義(F=2.69,P＜0.05).SDS-PAGE結果顯示,黃斑區脫離患者玻璃體中TTR四聚體結構明顯比黃斑裂孔患者和正常對照的穩定;對電泳條帶進行黑度積分分析,黃斑區脫離患者玻璃體中TTR含量約為黃斑裂孔患者玻璃體TTR含量的2.4倍,與ELISA的結果相一緻.CD結果顯示黃斑區脫離和黃斑裂孔患者玻璃體TTR的二級結構分彆含有45％α螺鏇和32％α螺鏇,且在不同pH(4～11)下保持穩定.正常樣本玻璃體TTR的二級結構含有5％α螺鏇,且隨pH(4～11)變化產生巨大波動.結論 黃斑區脫離患者以及黃斑裂孔患者玻璃體中TTR含量較高,二級結構相對穩定,且黃斑區脫離患者玻璃體TTR四聚體結構牢固,這可能與蛋白變異和錯誤摺疊有關.
목적 탐토전갑상선소단백(TTR)재병이성근시환자파리체중표체급기결구분석.방법 실험연구.수집황반구탈리(n=14)화병이성근시황반렬공(n=9)적파리체양본,령선정상각막공체안파리체양본(n=8)작위정상대조.대3조파리체진행료매련면역흡부시험(ELISA)、취병희선알응효전영(SDS-PAGE)、면역인적분석(western blot,WB)화원이색보(CD)분석.대소득수거진행단인소방차분석.결과 정상인、황반구탈리환자、황반렬공환자파리체중TTR적농도분별위(45.2±1.0)mg/L、(230.0±16.3)mg/L、( 109.8±19.0)mg/L,차이유통계학의의(F=2.69,P＜0.05).SDS-PAGE결과현시,황반구탈리환자파리체중TTR사취체결구명현비황반렬공환자화정상대조적은정;대전영조대진행흑도적분분석,황반구탈리환자파리체중TTR함량약위황반렬공환자파리체TTR함량적2.4배,여ELISA적결과상일치.CD결과현시황반구탈리화황반렬공환자파리체TTR적이급결구분별함유45％α라선화32％α라선,차재불동pH(4～11)하보지은정.정상양본파리체TTR적이급결구함유5％α라선,차수pH(4～11)변화산생거대파동.결론 황반구탈리환자이급황반렬공환자파리체중TTR함량교고,이급결구상대은정,차황반구탈리환자파리체TTR사취체결구뢰고,저가능여단백변이화착오절첩유관.
Objective To detect structures and levels of transthyretin (TTR) in the vitreous of pathological myopia patients.Methods It was a experimental study.TTR concentrations in the vitreous of pathological myopia patients complicated by macular detachment (MD) (n=14) and macular holes (MH) (n=9),and healthy controls (n=8) were analyzed by enzyme-linked immune sorbent assay (ELISA),sodium dodecyl sulfate gel electrophoresis (SDS-PAGE),Western Blot analysis and circular dichroism (CD) spectroscopy.The data abtained were analyzed by ANOVA.Results The concentration of TTR in the vitreous of healthy controls,macular detachment patients and macular hole patients was 45.2±1.0 mg/L,230.0±16.3 mg/L,109.8±19.0 mg/L,the difference was significant (F=2.69,P＜0.05).The transthyretin extracted from the vitreous of patients with MD showed a much more stable tetrameric structure than transthyretin extracted from the vitreous of patients with MH and the controls.Additionally,ELISA analysis and integrated optical density (IOD) analysis of bands on SDS-PAGE gels revealed that transthyretin concentration in the vitreous of patients with MD was 2.4 times higher than in patients with MH.CD spectroscopy revealed that the secondary structures of transthyretin in the vitreous of MD patients,MH patients and healthy,controls contained 45％,32％ and 5％ α-helix,respectively.Furthermore,the second structures of MD and MH transthyretin were stable at different pH levels (4-11),while that of healthy controls varied at pH levles 4-11.Conclusion Our results suggest that MD and MH patients have higher vitreous concentrations of transthyretin,and more stable secondary structures; additionally,MD TTR has a significantly steady tetrameric structure.These could possibly be due to mutations and misfolding.
