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目的研究视网膜近视因子基因在原代培养的豚鼠近视眼视网膜Müller细胞中的表达变化。方法3～4周龄断乳三色豚鼠40只,取20只建立近视眼模型(以右眼作为实验眼,以对侧未遮盖眼作自身对照),剩余的20只豚鼠作正常对照。用眼罩遮盖法建立豚鼠近视眼模型,用酶消化法原代培养其视网膜Müller细胞。RT-PCR检测酪氨酸羟化酶(tyrosine hydroxylase,TH)、诱导型一氧化氮合成酶(induciblenitric oxide synthase,iNOS)、神经型一氧化氮合成酶(neu-ronal nitric oxide synthase,nNOS)、内皮型一氧化氮合成酶(endothelial nitric oxide synthase,eNOS)、视黄醛脱氢酶(retinaldehyde dehydrogenase,RALDH)、醛脱氢酶(aldehydedehydrogenase,ALDH)、碱性成纤维细胞生长因子(basic fi-broblast growth factor,bFGF)、转化生长因子β(transforminggrowth factor-β,TGF-β)的mRNA在正常对照眼、自身对照眼和近视眼视网膜Müller细胞中的表达情况。数据分析采用One-way ANOVA检验。结果眼罩遮盖10d后,遮盖眼眼轴延长,近视形成。酶消化法成功培养出视网膜Müller细胞。正常对照眼、自身对照眼和遮盖眼视网膜Müller细胞均阳性表达nNOS、bFGF、TH、TGF-βmRNA,且遮盖眼表达nNOS、bFGF和TGF-βmRNA上调(P<0.05),TH mRNA下调(P<0.05)。iNOS mRNA仅在遮盖眼视网膜Müller细胞阳性表达。三组视网膜Müller细胞均不表达eNOS、RALDH和ALDHmRNA。结论豚鼠近视眼视网膜Müller细胞表达nNOS、iNOS、bFGF和TGF-βmRNA上调,TH mRNA下调。Müller细胞是近视眼视网膜信号因子一氧化氮(nitric oxide,NO)、多巴胺(dopamine,DA)、bFGF和TGF-β的一个重要来源。
Objective To study the changes of retinal myopia gene expression in primary cultured guinea pig myopic retinal Müller cells. Methods Totally 40 guinea pigs of weaning type were selected from 3 to 4 weeks old. Twenty eyes of 20 guinea pigs were established. The right eyes were used as the experimental eyes and the contralateral non-covered eyes were used as their own controls. The remaining 20 guinea pigs were used as controls. The model of guinea pig myopia was established by covering with eye mask, and the retinal Müller cells were primary cultured by enzyme digestion. RT-PCR was used to detect tyrosine hydroxylase (TH), inducible nitric oxide synthase (iNOS), neu-ronal nitric oxide synthase (nNOS) Endothelial nitric oxide synthase (eNOS), retinaldehyde dehydrogenase (RALDH), aldehyde dehydrogenase (ALDH), basic fibroblast growth factor (basic fi- (bFGF) and transforming growth factor-β (TGF-β) mRNA in normal control eye, self-control eye and myopic retinal Müller cells. Data analysis using One-way ANOVA test. Results eye mask cover 10d, cover the eye axis lengthening, myopia formation. Enzyme digestion of retinal Müller cells successfully cultured. The expression of nNOS, bFGF, TH and TGF-βmRNA in normal control eye, self-control eye and occluding retina Müller cells were upregulated (P < 0.05). iNOS mRNA is only expressed in retinal Müller cells. Three groups of retinal Müller cells did not express eNOS, RALDH and ALDH mRNA. Conclusion The expression of nNOS, iNOS, bFGF and TGF-βmRNA in guinea pig myopic retina Müller cells are up-regulated and the TH mRNA is down-regulated. Müller cells are an important source of nitric oxide (NO), dopamine (DA), bFGF and TGF-β in myopia.