玻璃体切割联合硅油填充术后晶状体基因表达谱的变化及验证

doi:10.6040/j.issn.1673-3770.0.2018.101

摘要/Abstract

摘要： 目的运用基因芯片技术研究玻璃体切割联合硅油填充术后晶状体的基因表达谱变化并进行验证。方法健康成年兔18只,随机分为术后3 d、1个月、3个月三组,每组6只,任一眼行玻璃体切割术后充填硅油,未手术眼作为对照。在不同的时间点各处死2只后取晶状体组织,应用基因芯片技术对晶状体的基因表达谱进行检测,和差异表达基因的筛选,并采用实时定量PCR技术对差异表达的基因进行验证。结果玻璃体切割硅油填充术后3 d与对照组相比差异表达基因共有129个,以炎症反应和氧化应激反应相关的基因为主;术后1个月与对照组相比差异表达基因共有140个,以细胞凋亡、物质转运相关的基因为主;硅油填充术后3个月与对照组相比差异表达基因共134个,以物质能量代谢、细胞分化增殖相关的基因为主。聚类分析显示,硅油填充术后3 d、1个月和3个月的实验组与对照组相比,共同表达变化的基因共有29个,其中共同表达上调的基因是NPPA、CD38、BPGM、 PIN、ELAM1等n个基因;共同表达下调的基因是VTN、GSTM2、BFSP2、LTB4DH、MMP2、CAPNS1等n个基因。应用real time PCR技术对这些表达变化关键基因的表达水平进行了进一步的验证,证实了表达谱芯片结果的真实性和可靠性。结论硅油填充术后晶状体基因表达谱涉及多基因改变,与炎症反应、氧化应激反应相关的基因的改变可能是术后形成晶状体混浊的始动因素。

关键词: 表达谱芯片, 差异表达基因, 晶状体, 硅油

Abstract: Objective To explore the significantly differentially expressed genes(DEGs)in lenses via microarray expression profiling after vitrectomy and silicone oil tamponade co-treatment. Methods After vitrectomy and silicone oil tamponade usage, mRNA expression profiling was carried out 3 days, 1 month, and 3 months after surgery in experimental rabbits. A non-surgery control group was also established and analyzed. Furthermore, genes with significantly altered expression after surgery were detected via quantitative real-time PCR. Results The expression of 129 genes was significantly altered 3 days after surgery and these included 55 upregulated and 74 downregulated genes(altered ratio, ≥ 2 or ≤ 0.5). The DEGs were mainly involved in the oxidative stress and inflammation regulatory pathways. Moreover, 140 genes were differentially expressed 1 month after the surgery, including 72 significantly upregulated and 68 significantly downregulated genes. Three months after surgery, 134 genes showed significantly altered expression, including 75 upregulated and 59 downregulated genes. Cluster analysis revealed 29 post-surgery DEGs common to the 3 experimental groups, including 10 upregulated and 9 downregulated genes. The representative upregulated genes included NPPA, CD38, BPGM, PIN, and ELAM1, and the representative downregulated genes included VTN, GSTM2, BFSP2, LTB4DH, MMP2, and CAPNS1. The significantly DEGs were involved in functions such as oxidative stress, inflammation, transport, signaling transduction, apoptosis, and cell differentiation. Furthermore, real-time PCR analysis confirmed the altered expression of these genes, indicating the authenticity and reliability of our microarray expression profiling analysis. Conclusion mRNA expression profiling revealed a group of genes significantly altered in lenses after vitrectomy and silicone oil tamponade co-treatment. These altered genes were involved in oxidative stress and inflammation regulatory pathways, which may be responsible for cataract formation.

Key words: Gene chip, Differentially expression gene, Lenses, Silicone oil

中图分类号:

R776

刘冬梅,李洋,宋继科,郭大东,马先祯,毕宏生. 玻璃体切割联合硅油填充术后晶状体基因表达谱的变化及验证[J]. 山东大学耳鼻喉眼学报, 2019, 33(2): 95-98.

LIU Dongmei, LI Yang, SONG Jike, GUO Dadong, MA Xianzhen, BI Hongsheng. Gene expression profile of lenses after vitrectomy and silicone oil tamponade co-treatment[J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2019, 33(2): 95-98.

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