体外培养牛眼小梁细胞表达胰岛素样生长因子-Ⅰ的研究

Expression of insulin-like growth factor-Ⅰ in bovine trabecular meshwork cells in vitro

目的 了解体外培养牛眼小梁细胞是否表达胰岛素样生长因子 Ⅰ (insulin likegrowthfactor Ⅰ ,IGF Ⅰ )。方法 体外培养新生牛眼小梁细胞并传 3代。采用Trizol试剂盒一步法提取细胞RNA ,应用高度特异性寡核苷酸引物对IGF Ⅰ的mRNA进行逆转录聚合酶链式反应 (reversetranscriptase polymerasechainreaction ,RT PCR)检测 ,并对扩增产物进行DNA双链测序鉴定。应用免疫组化Supervision染色法检测IGF Ⅰ蛋白的表达。结果 体外培养细胞符合牛眼小梁细胞的特点。RT PCR扩增得到单一的产物 ,其序列与已知序列完全一致。IGF Ⅰ免疫组化染色阳性。结论 小梁细胞自身产生IGF Ⅰ ,IGF Ⅰ的异常变化可能与原发性开角型青光眼的发病有关。

Objective To determine whether cultured bovine trabecular meshwork cells in vitro would express insulin like growth factor Ⅰ (IGF Ⅰ) Methods Newborn bovine trabecular meshwork cells of the third passage were cultured in vitro The whole RNA from 10 6 cells was extracted with Trizol reagent, and specific oligonucleotide primer pair and reverse transcription polymerase chain reaction (RT PCR) were used for detection of IGF Ⅰ messenger RNA Immunohistochemical stain was used to detect IGF Ⅰ protein Results Cultured cells had the specific characteristics of bovine trabecular meshwork cells A single RT PCR amplified product whose sequence was homologous to the known sequence was obtained IGF Ⅰ immunostain was positive Conclusions Trabecular meshwork cells may produce IGF Ⅰ This finding strongly supports the possibility that IGF Ⅰ may affect trabecular meshwork cells through paracrine and autocrine mechanisms