RBPJ对鸭胚成纤维细胞增殖和基因表达的影响机制

doi:10.3969/j.issn.1004-1524.20250649

浙江农业学报 ›› 2026, Vol. 38 ›› Issue (7): 1336-1344.DOI: 10.3969/j.issn.1004-1524.20250649

RBPJ对鸭胚成纤维细胞增殖和基因表达的影响机制

牛禹鸰^a(), 张羽洁^a, 贾渌雅^a, 赵阿勇^a, 刘长国^b, 杜雪^a^,^*()

^a 浙江农林大学动物科技学院·动物医学院,浙江杭州 311300
^b 浙江农林大学食品与健康学院(现代粮食产业学院),浙江杭州 311300

收稿日期:2025-10-17 出版日期:2026-07-25 发布日期:2026-08-20
作者简介:牛禹鸰,主要从事动物遗传育种与繁殖研究。E-mail:nyl@stu.zafu.edu.cn
通讯作者: *杜雪,E-mail: duxue@zafu.edu.cn
基金资助:
国家自然科学基金(32402739);浙江省畜禽农业新品种选育重大科技专项(2021C02068-10);国家级大学生创新创业训练计划项目(202410341011)

The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts

NIU Yuling^a(), ZHANG Yujie^a, JIA Luya^a, ZHAO Ayong^a, LIU Zhangguo^b, DU Xue^a^,^*()

^a College of Animal Science and Technology · College of Veterinary Medicine,Zhejiang A& F University,Hangzhou 311300,China
^b College of Food and Health (College of Modern Food Industry),Zhejiang A& F University,Hangzhou 311300,China

Received:2025-10-17 Published:2026-07-25 Online:2026-08-20

摘要/Abstract

摘要： 蛋鸭输卵管在产蛋过程中需要持续经历组织重塑、细胞更新和稳态维持,其中成纤维细胞作为主要间质细胞,在细胞外基质重塑、上皮分化支持及平滑肌功能维持中发挥基础作用。免疫球蛋白Kappa J区重组信号结合蛋白(recombination signal binding protein for immunoglobulin kappa J region,RBPJ)是Notch信号通路的重要效应因子,可通过调节细胞的增殖分化,影响组织发育、形态和功能。然而,RBPJ是否通过影响鸭输卵管成纤维细胞进而调控蛋鸭输卵管功能的研究尚未见报道。本研究通过细胞培养、RNA干扰(RNAi)和转录组学分析,探讨RBPJ对鸭成纤维细胞活力和增殖的影响及其对下游基因表达的调控作用。结果表明:(1)成功分离鸭胚成纤维细胞,并构建RBPJ干扰细胞模型;(2)RBPJ基因干扰后,鸭胚成纤维细胞活力(12 h)和增殖能力(24 h)显著(p<0.05)降低;(3)RBPJ基因干扰后引起364个基因差异表达,这些差异基因显著富集在MAPK和钙信号通路等信号通路中。综上,RBPJ基因通过调节PLA2G4C和AREG基因表达影响MAPK信号通路,通过调节FGF9、NTRK3和AGTR1基因表达影响钙信号通路,进而影响细胞增殖。研究结果弥补了RBPJ对鸭胚成纤维细胞增殖功能影响的研究空白,同时也为后续解析RBPJ对鸭输卵管成纤维细胞的功能奠定了研究基础,并在实际生产中为提高产蛋性能提供理论基础和科研支撑。

关键词: RBPJ基因, 鸭, 输卵管, 成纤维细胞, 细胞功能

Abstract:

During the egg-laying process, the oviduct of laying ducks must continuously undergo tissue remodeling, cell renewal, and homeostasis maintenance. Among these processes, fibroblasts, as the primary stromal cells, play a fundamental role in extracellular matrix remodeling, supporting epithelial differentiation, and maintaining smooth muscle function. The recombination signal binding protein for immunoglobulin kappa J region (RBPJ) is an important effector of the Notch signaling pathway, which regulates cell proliferation and differentiation, thereby affecting tissue development, morphology, and function. However, whether RBPJ regulates the function of duck oviduct by influencing duck oviduct fibroblasts has not yet been reported. This study investigated the effects of RBPJ on the viability and proliferation of duck embryo fibroblasts and its regulatory role in downstream gene expression through cell culture, RNA interference (RNAi), and transcriptome analysis. The results showed that: (1) Duck embryo fibroblasts were successfully isolated and RBPJ-interfered cell models were successfully constructed; (2) RBPJ gene interference significantly (p<0.05) reduced the viability at 12 h and the proliferation ability at 24 h of duck embryonic fibroblasts; (3) After RBPJ gene interference, 364 genes were differentially expressed and these significantly enriched in signaling pathways such as the MAPK and calcium signaling pathways. In conclusion: RBPJ affects the MAPK signaling pathway by regulating the expression of the PLA2G4C and AREG genes, and affects the calcium signaling pathway by regulating the expression of the FGF9, NTRK3, and AGTR1 genes, thereby affecting cell proliferation. These findings fill the research gap regarding the role of RBPJ in the proliferation of duck embryo fibroblasts. Moreover, they lay a research foundation for future analysis of RBPJ functions in duck oviduct fibroblasts and provide a theoretical basis and scientific support for improving egg-laying performance in practical production.

Key words: RBPJ gene, duck, oviduct, fibroblasts, cell function

中图分类号:

S834

牛禹鸰, 张羽洁, 贾渌雅, 赵阿勇, 刘长国, 杜雪. RBPJ对鸭胚成纤维细胞增殖和基因表达的影响机制[J]. 浙江农业学报, 2026, 38(7): 1336-1344.

NIU Yuling, ZHANG Yujie, JIA Luya, ZHAO Ayong, LIU Zhangguo, DU Xue. The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts[J]. Acta Agriculturae Zhejiangensis, 2026, 38(7): 1336-1344.

图/表 5

表1 siRNA序列

Table 1 siRNA sequences

siRNA名称 siRNA name	序列(5'→3') Sequence (5'→3')
NC-F	UUCUCCGAACGUGUCACGUTT
NC-R	ACGUGACACGUUCGGAGAATT
Si260-F	GCAGAAAUCAUAUGGAAAUTT
Si260-R	AUUUCCAUAUGAUUUCUGCTT
Si440-F	CUUGGAAGGAAAGAAUUAUTT
Si440-R	AUAAUUCUUUCCUUCCAAGTT

表2 引物序列

Table 2 Primer sequences

基因 Gene	正向引物序列(5'→3') Forward primer sequence (5'→3')	反向引物序列(5'→3') Reverse primer sequence (5'→3')	产物长度/bp Product length/bp
ACTB	AGCCATCTTTCTTGGGTATGGA	TTGTCACAAGGGTGTGGGTG	158
RBPJ	TAATCCAGTTTCAAGCCACTCC	CTTGGACTGGTTGACGGACC	165

图1 RBPJ干扰效果检测 A,RBPJ mRNA相对表达量,以ACTB为内参基因;B,RBPJ蛋白条带图;C,RBPJ蛋白灰度值相对表达量,以GAPDH为内参蛋白。NC,阴性对照siRNA;Si260和Si440,为靶向RBPJ的siRNA。柱上无相同小写字母表示差异显著(p<0.05);ns,差异不显著(p>0.05)。下同。

Fig.1 Detection of RBPJ knockdown efficiency A, Relative mRNA expression level of RBPJ, normalized to ACTB; B, Representative Western blot bands of RBPJ protein; C, Relative gray value quantification of RBPJ protein expression, normalized to GAPDH. NC, negative control siRNA; Si260 and Si440, siRNAs targeting RBPJ. Different lowercase letters above the bars indicate significant differences (p<0.05); ns, No significant difference (p>0.05). The same applies to the following figures.

图2 RBPJ干扰对成纤维细胞增殖能力和细胞活力的影响 A,EdU法检测细胞增殖(比例尺=100 μm)。Hoechst 33342染色标记细胞核(蓝色),EdU掺入标记S期细胞(红色),Merge图为叠加图像。B,EdU阳性细胞率统计。C,CCK-8法检测细胞活力。*代表NC组与Si440组之间存在显著(p<0.05)差异。

Fig.2 Effects of RBPJ siRNA on fibroblast proliferation and viability A, EdU assay was used to detect cell proliferation (scale bar=100 μm). Hoechst 33342 staining labeled the nuclei (blue), and EdU incorporation labeled S-phase cells (red), the Merge panels show the overlays of the two channels. B, Quantification of EdU-positive cell rates. C, Cell viability was measured by CCK-8 assay. * indicates a significant (p<0.05) difference between the NC group and the Si440 group.

图3 RBPJ干扰对成纤维细胞转录组表达谱的影响 A,主成分分析;B,差异表达基因火山图;C,差异表达基因的GO功能富集分析;D,差异表达基因的KEGG通路富集分析。

Fig.3 The effect of RBPJ interference on fibroblast transcriptome profiles A, Principal component analysis; B, Volcano plot of differentially expressed genes; C, GO functional enrichment analysis of differentially expressed genes; D, KEGG pathway enrichment analysis of differentially expressed genes.

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RBPJ对鸭胚成纤维细胞增殖和基因表达的影响机制

The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts

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