| ObjectiveEstablish the model of posterior capsule opacification(PCO) in rats. Detect andanalysis the difference of gene expression profile of lens epithelial cells(LECs) after extracapsular lens extraction(ECLE) in rats, and look for the molecular mechanism and new research clews of PCO. Look for a new molecular biology method to prevent PCO.Methods(1) An ECLE was performed in 12 consecutive Sprague Dawley(SD) rats atdouble eyes. Rats were sacrefied at 0 hour, 1, 7, and 14 days after surgery. Eyes were enucleated and processed for light microscopy. Variance analysis was used to compare the number of proliferative LECs.(2) 20 SD rats were divided into 4 groups at random and all received ECLE at double eyes. The rats were sacrificed at 0 hour, 1, 7, and 14 days after surgery and the capsules of lens were collected. The cDNA were retrotranscribed from the extracted RNA and the cy3- or cy5-labeled cRNA derived from the transcription of cDNA were fragmented as probes respectively. The probes were hybridized with rat oligo gene chip containing 5705 genes. Scanarray Scanner was used to screen the signals of hybridization and GenePix Pro 4.0 software was applied to analysis the expression of genes. The expressed quantity of Bfsp1, Ccnd1, Cdk4, Cdkn1b and Cdkn2b were detected using quantitative real-time polymerase chitin reaction (qRT-PCR) to certify the results of genechips.(3) 48 SD rats were divided into 3 groups at random and all received ECLE at double eyes, Ccnd1 antisense oligonucleotides(Ccnd1-ASON) liposomes, Ccndl sense oligonucleotides(Ccnd1-SON) liposomes and liposome without any drugs were respectively injected into anterior chamber of different groups. 4 rats of each group were sacrefied at 0 hour, 1, 7, and 14 days after surgery and the eyes were enucleated. 4 eyes used to detect the expression of Ccnd1 using qRT-PCR. The other 4 eyes wereprocessed for light microscopy and the expression of cyclin Dl in LECs examined by immunohistochemical technique. Variance analysis was used to compare the number of proliferative LECs and the integrated optical density(IOD) of cyclin Dl positive LECs.Results(1) All eyes got successfully ECLE. Immediately after surgery, single layer ofLECs were present in the inner surface of the anterior capsule and lens bow;1 day after surgery, LECs started to migrate toward the center of the posterior capsule;at 7 days, multiple layers of LECs were emerged at the sufface of the posterior capsule and lens bow, capsular wrinkling was apparent;the mass of LECs, lens fibers and Soemmering's ring were observed in all eyes 14 days. The number of LECs increases obviously as time goes(p<0.05).(2) The quality control and the results of chips was reliable. There were 351(6.2%), 252(4.4%), 367(6.4%) differentially expressed genes at the 3 points after ECLE respectively using a two-fold criterion, among them 196,120,171 increased and 155,132,196 decreased. Integrated 3 points, 694 genes expressed differentially. These differentially expressed genes encompassed all functional classes.(3) The espression quantity of Ccndl in group of Ccndl-ASON was larger than the other two groups which got no difference. The numbers of LECs and espression level of cyclin Dl of LECs in group of Ccndl-ASON were higher than the other two groups(p<0.05) while the other two got no difference.Conclusions(1) Using ECLE, the model of PCO can be successfully established in rats.(2) The differential genes expression of LECs after ECLE in rats was intricate which indicate the molecular mechanism of PCO was complicated and multiplex regulative factors may involve in at the same time.(3) Anterior chamber injecting of Ccndl-ASON liposomes had inbiting effects on proferation of LECs and reduced PCO in some degree. |
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