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蛋白激酶C活性调节剂对豚鼠近视眼视网膜Mller细胞的作用及意义
  • ISSN号:1003-0808
  • 期刊名称:眼科研究
  • 时间:0
  • 页码:3222-3226
  • 语言:中文
  • 分类:R774.03[医药卫生—眼科;医药卫生—临床医学]
  • 作者机构:[1]中南大学湘雅医院眼科,长沙410008
  • 相关基金:本课题为国家自然科学基金青年基金资助(30600694)
  • 相关项目:Müller细胞调控豚鼠近视眼视网膜信号因子的研究
中文摘要:

目的研究蛋白激酶C(PKC)激活剂PMA和抑制剂GF109203X对豚鼠近视眼视网膜Mailer细胞生长状态及PKC蛋白表达的影响。方法眼罩遮盖诱导近视模型,酶消化法培养其视网膜Mailer细胞,并鉴定。PMA、GF109203X分别干预近视眼Mailer细胞。MTT测细胞活力,TUNEL染色检测凋亡细胞,Westren blotting测PKC蛋白表达。结果95%以上的培养细胞阳性表达胶质纤维酸性蛋白(GFAP)和波形蛋白(Vimentin)。近视组PKC蛋白表达较正常组上调(P〈0.05)。PMA、GF109203X均能引起近视眼Mailer细胞的抑制率升高和诱导其凋亡(P〈0.05)。PMA诱导近视眼Mailer细胞PKC蛋白表达进一步上调,100nmol/L组和1000nmol/L组表达量高于10nmol/L组(P〈0.05)。1μmol/L和10μmol/L GF109203X均能引起近视眼Mailer细胞PKC蛋白表达下调,10μmol/L的抑制作用大于1μmol/L(P〈0.05)。结论PKC蛋白在豚鼠近视眼视网膜Mailer细胞表达升高,且其表达受PMA和GF109203X调控。

英文摘要:

Objective To investigate the effect of protein kinase C (PKC) activitor,PMA,and PKC inhibitor,GF109203X,on the growth state and PKC protein in the primary cultured retinal Mailer ceils of myopic eye in guinea pig. Methods The form- deprived myopia model was induced in 30 guinea pigs by translucent goggles. Retinal Mailer cells from myopia model were isolated and cultured using enzyme-digesting method and identified by glial fibrillary acidic protein(GFAP) and Vimentin stain. Cultured retinal Mailer cells were divided into normal control group,myopia group, myopia plus PMA 10 nmol/L group, myopia plus PMA 100 nmol/L group,myopia plus PMA 1 000 nmol/L group, myopia plus GF109203X 0. 1μmol/L group, myopia plus GF109203X 1 μmol/L group and myopia plus GF109203X 10 μmol/L group. Cell vitality was evaluated by MTT colorimetry, and the apoptotic Mailer cells were detected by one-step TUNEL staining. The expression of PKC protein in cultured retinal Miiller cells was analyzed by Westren blotting. Results After 14 days,the occluded eyes became myopic in guinea pigs. Over 95% cultured cells showed positive reaction for GFAP and Vimentin. Compared with normal control group, the expression of PKC protein was significantly increased in myopia group(P 〈0.05) ,and that in myopia plus 100 nmol/L PMA and myopia plus 1 000 nmol/L PMA groups was stonger than in 10 nmol/L PMA group(P 〈 0. 05 ). No statistically significant change was found between 100 nmol/L PMA group and 1 000 nmol/L PMA group(P 〉0. 05). The expression of PKC protein in myopic Mailer cells was significantly decreased in 1 μmol/L and 10 μmol/L GF109203X groups,and a statistically significant enhance was seen in 10 μmol/L GF109203X group compared with 1 μmol/L GF109203X group (P 〈 0. 05). Conclusion PKC protein may be involved in the formation of form-deprived myopia. The level of PKC protein in retinal Mailer cells is regulated by PMA and GF109203X to certain degree.

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