视网膜内源性多肽CKASKGKL调节p53/cyclinD1信号通路拮抗多氯联苯暴露致子代视网膜损伤-东南大学学报医学版

视网膜内源性多肽CKASKGKL调节p53/cyclinD1信号通路拮抗多氯联苯暴露致子代视网膜损伤

单位：1. 南京医科大学附属妇产医院儿童保健科, 江苏南京 210004;
2. 宁波市第一医院新生儿科, 浙江宁波 315010;
3. 南京医科大学附属妇产医院医学研究中心, 江苏南京 210004

关键词：内源性多肽多氯联苯视网膜发育 p53/cyclinD1信号通路斑马鱼大鼠

分类号：R774.1

出版年·卷·期（页码）：2023·42·第一期（15-21）

摘要：

目的：探讨视网膜内源性多肽CKASKGKL对多氯联苯（PCBs）暴露致子代视网膜发育异常的拮抗作用及可能的作用机制。方法：以斑马鱼为模式动物，收集斑马鱼胚胎予以PCBs处理，同时给予多肽CKASKGKL进行挽救实验，收集受精后48 h幼鱼制备石蜡切片，显微镜下观察各组子代斑马鱼视网膜发育情况。以大鼠视网膜神经节细胞（RGC）为工具细胞，予以PCBs处理，同时给予多肽CKASKGKL进行挽救实验，CCK-8检测各组细胞增殖活性；采用基因富集分析（GSEA）了解多肽CKASKGKL可能作用的信号通路，Western blotting初步验证多肽CKASKGKL对视网膜神经节细胞p53/cyclinD1信号通路的影响。结果：多肽CKASKGKL可以拮抗PCBs暴露致子代斑马鱼视网膜发育异常，可以拮抗PCBs暴露致视网膜神经节细胞增殖活性降低；GSEA结果显示，多肽CKASKGKL可能通过参与p53信号通路发挥作用；Western blotting检测显示，多肽CKASKGKL干预后，视网膜神经节细胞中p53表达降低，其下游负调控基因cyclinD1表达增高。结论：多肽CKASKGKL可以拮抗PCBs暴露导致的子代视网膜损伤，可能与其通过调节p53/cyclinD1信号通路而促进细胞增殖有关。

Objective: To explore the antagonistic effect of endogenous retinal peptide CKASKGKL on offspring retinal dysplasia induced by PCBs and its possible mechanism. Methods: Zebrafish were selected as model animal, embryos were collected and treated with PCBs, and CKASKGKL was treated to perform rescue experiment simultaneously, the juveniles were collected to make paraffin sections 48 hours after fertilization, and the retina of zebrafish offspring in each group was observed under microscope. Rat retinal ganglion cells(RGCs) were used as tool cells, treated with PCBs, at the same time treated with CKASKGKL for rescue experiment, and CCK-8 was used to detect the proliferation activity of cells in each group. Gene enrichment analysis(GSEA) method was used to analyze the signaling pathways in which CKASKGKL might function, Western blotting was used to verify the effect of CKASKGKL on the p53/cyclinD1 signaling pathway in RGCs preliminarily. Results: CKASKGKL could antagonize the retinal dysplasia of zebrafish offspring caused by PCBs, and could antagonize the decrease of proliferation activity of RGCs caused by PCBs exposure. The results of GSEA showed that CKASKGKL might play a role through p53 signaling pathway. The results of Western blotting demonstrated that after treated with CKASKGKL, the expression of p53 in RGCs decreased, and the expression of its downstream negatively regulated gene cyclinD1 increased. Conclusion: CKASKGKL can antagonize the offsprings retinal damage caused by PCBs exposure, which may be related to the promotion of cell proliferation through the p53/cyclinD1 signaling pathway.

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