Nicotinamide improves in vitro lens regeneration in a mouse capsular bag model.
Nicotinamide improves in vitro lens regeneration in a mouse capsular bag model.
复制标题
烟酰胺改善小鼠囊袋模型的体外晶状体再生
DOI：
10.1186/s13287-022-02862-8
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发表时间：
2022-05-12
影响因子：
7.5
通讯作者：
中科院分区：
文献类型：
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作者：
背景哺乳动物透镜再生在白内障治疗中具有巨大的潜力。然而，哺乳动物透镜再生的机制尚不清楚，优化的方法仍然存在问题。MethodsWe开发了一种体外透镜再生模型，使用小鼠囊袋培养，并提高了再生透镜的透明度，使用烟酰胺（NAM）。我们分别使用D4476和SSTC 3作为酪蛋白激酶1A抑制剂和激动剂。晶状体特异性标记物的表达通过实时PCR、免疫染色和蛋白质印迹进行检查。采用3，3 ′-二己基氧杂碳菁碘化物（DiOC 6）和亚甲蓝染色、末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）和透射电镜等方法对体外再生晶状体的结构进行了研究。在早期阶段，晶状体透镜上皮细胞穿过后囊增殖并分化为透镜纤维细胞（LFC）。再生晶状体在28天后出现不透明;然而，NAM处理有效地保持了再生透镜的透明度。我们证明，NAM维持透镜上皮细胞的存活，促进LFC的分化和规则的细胞排列，并减少晶状体相关的细胞凋亡。机械地，NAM增强了分化和透明的再生晶状体部分通过抑制酪蛋白激酶1A activity.ConclusionThis研究提供了一个新的体外模型再生研究，并证明了潜在的NAM在体外哺乳动物透镜再生。
BackgroundMammalian lens regeneration holds great potential as a cataract therapy. However, the mechanism of mammalian lens regeneration is unclear, and the methods for optimization remain in question.MethodsWe developed an in vitro lens regeneration model using mouse capsular bag culture and improved the transparency of the regenerated lens using nicotinamide (NAM). We used D4476 and SSTC3 as a casein kinase 1A inhibitor and agonist, respectively. The expression of lens-specific markers was examined by real-time PCR, immunostaining, and western blotting. The structure of the in vitro regenerated lens was investigated using 3,3′-dihexyloxacarbocyanine iodide (DiOC6) and methylene blue staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and transmission electron microscopy.ResultsThe in vitro lens regeneration model was developed to mimic the process of in vivo mammalian lens regeneration in a mouse capsular bag culture. In the early stage, the remanent lens epithelial cells proliferated across the posterior capsule and differentiated into lens fiber cells (LFCs). The regenerated lenses appeared opaque after 28 days; however, NAM treatment effectively maintained the transparency of the regenerated lens. We demonstrated that NAM maintained lens epithelial cell survival, promoted the differentiation and regular cellular arrangement of LFCs, and reduced lens-related cell apoptosis. Mechanistically, NAM enhanced the differentiation and transparency of regenerative lenses partly by inhibiting casein kinase 1A activity.ConclusionThis study provides a new in vitro model for regeneration study and demonstrates the potential of NAM in in vitro mammalian lens regeneration.
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影响因子：
64.8
作者：
通讯作者：
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影响因子：
--
作者：
Henry, Jonathan J.;Thomas, Alvin G.;Hamilton, Paul W.;Moore, Lisa;Perry, Kimberly J.
通讯作者：
Perry, Kimberly J.
影响因子：
3.7
作者：
Griffin SM;Pickard MR;Orme RP;Hawkins CP;Williams AC;Fricker RA
通讯作者：
Fricker RA
影响因子：
16
作者：
Avalos, JL;Bever, KM;Wolberger, C
通讯作者：
Wolberger, C
影响因子：
64.5
作者：
Sachs, Norman;de Ligt, Joep;Clevers, Hans
通讯作者：
Clevers, Hans