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胰岛素样生长因子-I基因在牛小梁网组织和细胞中的克隆及蛋白表达

Insulin-like growth factor-I gene cloning and protein expression in bovine trabecular meshwork tissue and cells.

作者信息

Cao Yang, Wei Houren, Da Banghong, Pfaffl Michael, Li Zhongyu

机构信息

Department of Ophthalmology, Xiehe Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022.

出版信息

J Huazhong Univ Sci Technolog Med Sci. 2002;22(1):69-72. doi: 10.1007/BF02904794.

Abstract

Whether cultured bovine trabecular meshwork cells and trabecular tissue ex vivo express insulin-like growth factor-I (IGF-I) messenger RNA (mRNA) and protein was investigated. Total RNA of cultured bovine trabecular meshwork cells as well as trabecular meshwork tissue freshly excised from bovine eyes was extracted, and reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect IGF-I mRNA. RT-PCR product was verified by sequencing. Immunohistochemical stain was used to detect IGF-I protein. The results showed that a single PCR amplified product was obtained, and the sequence was homologous to the known sequence.. IGF-I immunostain was positive in the cytoplasm of trabecular meshwork cells. It was concluded that trabecular meshwork cells produce IGF-I and contribute to the presence of IGF-I in trabecular meshwork microenvironment as well as aqueous humor. Trabecular meshwork cells were affected by IGF-I not only through paracrine, but also autocrine action. Whether abnormal down-regulations in IGF-I production may contribute to the pathogenesis of primary open-angle glaucoma and the possibility of promoting the autocrine action of IGF-I by trabecular meshwork cells to treat the disease is worth further investigation.

摘要

研究了体外培养的牛小梁网细胞和小梁组织是否表达胰岛素样生长因子-I(IGF-I)信使核糖核酸(mRNA)和蛋白质。提取了培养的牛小梁网细胞以及从牛眼新鲜切除的小梁网组织的总RNA,并用逆转录聚合酶链反应(RT-PCR)检测IGF-I mRNA。通过测序验证RT-PCR产物。采用免疫组织化学染色检测IGF-I蛋白。结果显示获得了单一的PCR扩增产物,且序列与已知序列同源。IGF-I免疫染色在小梁网细胞的细胞质中呈阳性。结论是小梁网细胞产生IGF-I,并导致小梁网微环境以及房水中存在IGF-I。小梁网细胞不仅通过旁分泌,而且通过自分泌作用受IGF-I影响。IGF-I产生的异常下调是否可能导致原发性开角型青光眼的发病机制,以及小梁网细胞促进IGF-I自分泌作用以治疗该疾病的可能性,值得进一步研究。

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