Power W J, Kaufman A H, Merayo-Lloves J, Arrunategui-Correa V, Foster C S
Rhoads Molecular Immunology, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, USA.
Curr Eye Res. 1995 Oct;14(10):879-86. doi: 10.3109/02713689508995127.
A semi-quantitative polymerase chain reaction (PCR) methodology was used to evaluate the kinetic changes occurring in collagens I, III, IV and V mRNA in rat cornea following excimer laser keratectomy. cDNA was synthesized from RNA extracted from rat cornea at various times following excimer laser photoablative keratectomy. Collagen cDNA sequences were subsequently amplified using specific sets of oligonucleotide primers. Competitive PCR amplification was carried out using an internal standard so that a semi-quantitative analysis of message for synthesis of collagen types I, III, IV and V could be performed and time course dynamics of message for these collagens studied. There was a biphasic increase in the levels of collagens III, IV and V mRNA following excimer laser keratectomy. Collagen I mRNA levels demonstrated a more sustained increase and were still elevated at 6 weeks following wounding. Collagens IV and V mRNA showed the largest increase with an approximate three fold increase over controls between 4 days and 1 week. Our results demonstrate that upregulation of stromal collagens I, III, and V mRNA and basement membrane collagen IV mRNA occurs in rat cornea following excimer laser keratectomy.
采用半定量聚合酶链反应（PCR）方法评估准分子激光角膜切削术后大鼠角膜中I、III、IV和V型胶原mRNA的动态变化。在准分子激光光蚀性角膜切削术后不同时间，从大鼠角膜提取的RNA合成cDNA。随后使用特定的寡核苷酸引物组扩增胶原cDNA序列。使用内标进行竞争性PCR扩增，以便对I、III、IV和V型胶原合成的信息进行半定量分析，并研究这些胶原信息的时间进程动态。准分子激光角膜切削术后，III、IV和V型胶原mRNA水平呈双相增加。I型胶原mRNA水平显示出更持续的增加，在受伤后6周仍升高。IV和V型胶原mRNA增加最大，在4天至1周之间比对照组增加约三倍。我们的结果表明，准分子激光角膜切削术后大鼠角膜中基质胶原I、III和V型mRNA以及基底膜胶原IV型mRNA上调。
