不同蕈毒類藥劑對抑制近視眼球生長的影響（３／３）
Date Issued
2002
Date
2002
Author(s)
施永豐
DOI
902314B002184
Abstract
The prevention and treatment of myopia are important issue of public health in many countries, especially in Taiwan where the prevalence rate of myopia is extremely high. The most important complication of extreme myopia is retinal degeneration affecting the posterior pole that is associated with elongation of ocular axial length. Unfortunately, the actual mechanism of the development of myopia is still unknown
Various pharmacological agents have been tried to treat myopia. Anti-cholinergic agnets, such as atropine and pirenzepine were reported to be effective to prevent the progression of myopia. However the actual mechanism of these agent is still unknown. We hypothesized that atropine and pirenzepine could prevent the progression of myopia through influencing the expression of growth factors in retina-RPE-choroidal complex and sclera. In this project, the effect of 1% atropine and 2% pirenzepine on the expression of growth factor mRNA from the retina-RPE-choroidal complex and sciera will be examined in the chick model of form deprivation myopia.
Subtraction-hybridization PCR method is used to selectively amplify target
cDNA fragments and simultaneously suppress nontarget DNA amplification (45, 46). It can achieve greater than 1000-fold enrichment of differentially expressed cDNAs (ie. cDNA from myopic and control eyes). The basic idea of
subtraction-hybridization PCR is those tracers DNA (in our experiment, the cDNA from myopic eyes) will primarily reassouciate with excess driver DNA (in our
experiment, the cDNA from control eyes) while target sequences having no counterparts in driver will inevitably reassociate with each other, or remain
single-stranded. The reassociated fragments common for driver and tracer are discarded, and the remaining DNA enriched in target sequences is cloned and
analyzed. With this method, Ishibashi et al. showed the upreglation of crystalline mRNAs in form-deprived chick eyes (47). Den Hollander and coworker used this technique to isolate and map the novel candidate genes for retinal disorders (48). It
is proven that subtraction-hybridization PCR is a powerful tool to study chick myopia.
chloroform, shaking for 15 minutes, then cooling at 4¢XC for 5 minutes. The suspension was centrifuged for 15 minutes at 4¢XC and the aqueous phase transferred to a new tube. The RNA was precipitated by adding 600 t1 isopropanol, incubating on ice for 15 minutes, and centrifuging for 15 minutes at 4¢XC. The RNA pellets were washed once with I ml 75 % ethanol, dried, resuspended in 20 il diethyl pyrocarbonate (DEPC)-treated water and incubated for 10 minutes at 60¢XC. The RNA was stored frozen at -80¢XC. RNA purity was determined by measuring the 0D260/0D280 and RNA quantity was estimated from OD260.
SDGs
Publisher
臺北市：國立臺灣大學醫學院眼科
Type
journal article
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