Kawaji Takahiro, Ando Yukio, Nakamura Masaaki, Yamamoto Keiichi, Ando Eiko, Takano Akiomi, Inomata Yasuya, Hirata Akira, Tanihara Hidenobu
Department of Ophthalmology and Visual Science, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
Exp Eye Res. 2005 Sep;81(3):306-12. doi: 10.1016/j.exer.2005.02.003.
Ocular symptoms of transthyretin (TTR)-related familial amyloidotic polyneuropathy (FAP) suggest that ciliary pigment epithelium (CPE) may synthesize TTR and its TTR may lead to amyloid formation in addition to TTR from vessels and retinal pigment epithelium (RPE). To clarify sites of TTR synthesis in ocular tissues, we performed in situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR) for qualitative detection of TTR mRNA. In addition, we quantified levels of TTR mRNA expression by means of real-time quantitative RT-PCR. Furthermore, although TTR is an anti-acute phase protein in serum level, no reports on changes in TTR expression in ocular tissues during acute inflammation exist. To investigate changes in TTR expression in ocular tissues during inflammation, we induced uveitis by endotoxin challenge in rabbits and used real-time quantitative RT-PCR to examine changes in TTR mRNA expression in ocular tissues. In situ hybridization and RT-PCR qualitatively demonstrated TTR mRNA not only in RPE but also in CPE. Real-time quantitative RT-PCR showed that the level of TTR mRNA expression in the CPE was about one-third of that in the RPE. TTR mRNA expression in ocular tissues decreased as the degree of inflammation increased. These results suggest that TTR synthesized in the CPE may lead to ocular manifestations, especially glaucoma, in FAP. TTR mRNA also acts as an anti-acute phase reactant in ocular tissues.
转甲状腺素蛋白(TTR)相关家族性淀粉样多神经病(FAP)的眼部症状表明,睫状体色素上皮(CPE)可能合成TTR,并且其TTR除了来自血管和视网膜色素上皮(RPE)的TTR外,可能导致淀粉样蛋白形成。为了阐明眼部组织中TTR的合成部位,我们进行了原位杂交和逆转录聚合酶链反应(RT-PCR)以定性检测TTR mRNA。此外,我们通过实时定量RT-PCR对TTR mRNA表达水平进行定量。此外,尽管TTR在血清水平上是一种抗急性期蛋白,但尚无关于急性炎症期间眼部组织中TTR表达变化的报道。为了研究炎症期间眼部组织中TTR表达的变化,我们通过内毒素攻击在兔中诱导葡萄膜炎,并使用实时定量RT-PCR检测眼部组织中TTR mRNA表达的变化。原位杂交和RT-PCR定性显示TTR mRNA不仅存在于RPE中,也存在于CPE中。实时定量RT-PCR显示CPE中TTR mRNA表达水平约为RPE中的三分之一。眼部组织中TTR mRNA表达随炎症程度增加而降低。这些结果表明,CPE中合成的TTR可能导致FAP中的眼部表现,尤其是青光眼。TTR mRNA在眼部组织中也作为一种抗急性期反应物起作用。