Effects of Low-dose Triamcinolone Acetonide on Rat Retinal Progenitor Cells under Hypoxia Condition
Effects of Low-dose Triamcinolone Acetonide on Rat Retinal Progenitor Cells under Hypoxia Condition
复制标题
小剂量曲安奈德对缺氧条件下大鼠视网膜祖细胞的影响
DOI：
10.4103/0366-6999.184474
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发表时间：
2016
影响因子：
6.1
通讯作者：
Q. Kang
中科院分区：
文献类型：
--
作者：
Y. Xing;Li;Q. Kang
背景：视网膜退行性疾病是发达国家致盲的主要原因。视网膜祖细胞（Retinal progenitor cells, RPCs）在视网膜修复中起着关键作用。曲安奈德（Triamcinolone acetonide， TA）广泛用于视网膜退行性疾病的治疗。在本研究中，我们探讨了TA在缺氧条件下对RPCs的作用。方法：原代培养，免疫荧光染色鉴定。在TA处理条件下，细胞分别在常氧、缺氧6 h和缺氧6 h下培养。TA处理组在缺氧条件下培养6 h后，用不同浓度TA处理RPCs 48 ~ 72 h，采用细胞计数试剂盒-8 （CCK-8）法测定细胞活力。流式细胞术检测细胞周期。Western blot检测细胞周期蛋白D1、Akt、p-Akt、核因子(NF)-&kgr的表达；bp65和caspase-3。结果：CCK-8检测显示，与对照组相比，缺氧组0.01 mg/ml TA处理48 h后，RPCs活力明显提高（P < 0.05）。72h后，与对照组相比，0.01 mg/ml和0.02 mg/ml TA组细胞活力均有所提高（均P < 0.05）。流式细胞术显示，缺氧6 h组s期细胞数量明显多于常氧对照组（P < 0.05）。与其他各组相比，给予TA组S期和G2/M期的RPCs降低（均P < 0.05）。各组大鼠总Akt蛋白表达差异无统计学意义，p-Akt和NF-&kgr表达上调；与缺氧6 h组和对照组比较，0.01 mg/ml TA组B p65蛋白表达下调，caspase-3、cyclin D1蛋白表达下调（均P < 0.05）。结论：小剂量TA对RPCs有抗凋亡作用，但对细胞增殖无刺激作用。
Background:Retinal degenerative diseases are the leading causes of blindness in developed world. Retinal progenitor cells (RPCs) play a key role in retina restoration. Triamcinolone acetonide (TA) is widely used for the treatment of retinal degenerative diseases. In this study, we investigated the role of TA on RPCs in hypoxia condition. Methods:RPCs were primary cultured and identified by immunofluorescence staining. Cells were cultured under normoxia, hypoxia 6 h, and hypoxia 6 h with TA treatment conditions. For the TA treatment groups, after being cultured under hypoxia condition for 6 h, RPCs were treated with different concentrations of TA for 48–72 h. Cell viability was measured by cell counting kit-8 (CCK-8) assay. Cell cycle was detected by flow cytometry. Western blotting was employed to examine the expression of cyclin D1, Akt, p-Akt, nuclear factor (NF)-&kgr;B p65, and caspase-3. Results:CCK-8 assays indicated that the viability of RPCs treated with 0.01 mg/ml TA in hypoxia group was improved after 48 h, comparing with control group (P < 0.05). After 72 h, the cell viability was enhanced in both 0.01 mg/ml and 0.02 mg/ml TA groups compared with control group (all P < 0.05). Flow cytometry revealed that there were more cells in S-phase in hypoxia 6 h group than in normoxia control group (P < 0.05). RPCs in S and G2/M phases decreased in groups given TA, comparing with other groups (all P < 0.05). There was no significant difference in the total Akt protein expression among different groups, whereas upregulation of p-Akt and NF-&kgr;B p65 protein expression and downregulation of caspase-3 and cyclin D1 protein expression were observed in 0.01 mg/ml TA group, comparing with hypoxia 6 h group and control group (all P < 0.05). Conclusion:Low-dose TA has anti-apoptosis effect on RPCs while it has no stimulatory effect on cell proliferation.