北京大学学报(医学版) ›› 2014, Vol. 46 ›› Issue (1): 5-11.

• 论著 • 上一篇    下一篇

激活Notch信号通路可延迟人牙髓细胞的老化表现

常思佳,邹晓英,庄姮△,岳林,高学军   

  1. (北京大学口腔医学院·口腔医院牙体牙髓科,北京100081)
  • 出版日期:2014-02-18 发布日期:2014-02-18

Notch activation delayed ageing of human dental pulp cells

CHANG Si-jia, ZOU Xiao-ying, ZHUANG Heng△, YUE Lin, GAO Xue-jun   

  1. (Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology, Beijing 100081, China)
  • Online:2014-02-18 Published:2014-02-18

摘要: 目的:建立Notch信号通路激活的人牙髓细胞模型,探讨激活Notch信号通路对人牙髓细胞老化的影响。方法:体外培养人牙髓细胞,从第4代开始实验,将牙髓细胞分为激活组和阴性对照组。激活组采用10 mg/L的Jagged1蛋白包被培养皿的方法激活Notch信号通路,阴性对照组的培养皿不做处理。以实时定量PCR(real-time quantitative PCR,RT-qPCR)检测第4代、第8代、第10代Notch信号通路下游基因Hes1的表达水平。分三个水平观察Notch信号通路激活后人牙髓细胞的变化:(1)细胞水平,用倒置显微镜观察细胞形态的变化,用MTT法检测细胞活力改变;(2)代谢酶水平,用碱性磷酸酶(alkaline phosphatase, ALP)染色及活性测定试剂盒检测ALP的表达和活性,用衰老相关β-半乳糖苷酶染色方法检测阳性细胞百分率;(3)基因水平,用RT-qPCR检测老化相关基因p53和p16的表达。t检验比较激活组和阴性对照组各指标的差异。结果:采用Jagged1蛋白包被培养皿后,Notch信号通路下游基因Hes1表达增高,表明建立了Notch信号通路激活的人牙髓细胞模型。激活组牙髓细胞与阴性对照组牙髓细胞相比,较晚出现形态显现衰老的细胞,第8代、第10代细胞活力升高,ALP表达量升高;第10代ALP活性升高,β-半乳糖苷酶表达下降;各代p16基因表达水平均降低,第8代、第10代p53基因表达水平降低。结论:Jagged1蛋白可有效激活Notch信号通路,激活Notch信号通路后,人牙髓细胞在细胞水平、代谢酶水平、基因水平均表现出老化延迟的趋势。

关键词: 受体, Notch, 信号传导, 细胞衰老, 牙髓

Abstract: Objective:To establish model of dental pulp cells with activated Notch signaling pathway, and investigate the effect of activating Notch signaling pathway on senescence of human dental pulp cells in vitro. Methods: Human dental pulp cells were isolated, cultured as usual, and used from the 4th passage. The cells were divided into the activated group and the negative control group. In the activated group, the way of coating dishes with Jagged1 protein (10 mg/L) was used to activate Notch signaling pathway. The negative control group cells received no treatment. In the 4th, 8th, and 10th passages, the expression levels of the Notch signaling pathway downstream gene Hes1 were verified by real-time quantitative PCR (RT-qPCR). The cell changes after activating Notch signaling pathway were observed at three levels: (1) The cell morphology changes were observed through invert phase contrast microscope. The cell activity was detected with MTT assay. (2) The alkaline phosphatase (ALP) expression and its activity, and senescence-associated β-galatosidase (SA-β-Gal) expression were observed with the kit. (3) The expression changes of senescence related genes were verified using RT-qPCR. The difference between the negative control group and the activated group was analyzed using student’s t test. Results: The expression level of the downstream gene Hes1 of Notch signaling pathway increased after coating the dishes with Jagged1 protein, indicating the establishment of the model of dental pulp cells with activated Notch signaling pathway. Compared with the negative control group, the aging cells of the activated group appeared relatively late. In the 8th and 10th passage, the cell activity increased. In the 10th passage, ALP activity increased, but SA-β-Gal expression decreased. p16 gene expression decreased in each passage, and p53 gene expression decreased in the 8th and 10th passages. Conclusion: Jagged1 could activate Notch signaling pathway effectively. Through activating Notch signaling pathway, the dental pulp cells showed a trend of senescence delay at different levels, such as cell morphology, metabolic enzyme expressions and related gene expressions.

Key words: Receptors, Notch, Signal transduction, Cell aging, Dental pulp

[1] 赵晓一,刘畅,钱锟,潘洁. 成熟恒牙牙髓切断术的疗效及影像学评价[J]. 北京大学学报(医学版), 2024, 56(1): 138-143.
[2] 郑佳佳,杨雪,温泉,付元,邵校,丁美丽. 生物活性陶瓷iRoot BP Plus®在儿童年轻恒前牙复杂冠折牙髓切断术中的应用[J]. 北京大学学报(医学版), 2024, 56(1): 179-184.
[3] 扶琼,叶霜. 嵌合抗原受体T细胞治疗在自身免疫疾病中的应用和思考[J]. 北京大学学报(医学版), 2023, 55(6): 953-957.
[4] 叶雨阳,岳林,邹晓英,王晓燕. 成牙本质方向分化牙髓干细胞外泌体形态及微小RNA表达谱特征[J]. 北京大学学报(医学版), 2023, 55(4): 689-696.
[5] 代云飞,刘鹤,彭楚芳,姜玺军. 年轻恒牙牙髓再生治疗术后36个月的临床疗效评估[J]. 北京大学学报(医学版), 2023, 55(4): 729-735.
[6] 朱晓娟,张虹,张爽,李东,李鑫,徐玲,李挺. 人表皮生长因子受体2低表达乳腺癌的临床病理学特征及预后[J]. 北京大学学报(医学版), 2023, 55(2): 243-253.
[7] 叶佳学,梁宇红. 牙髓专科医师应用锥形束CT的现况调查[J]. 北京大学学报(医学版), 2023, 55(1): 114-119.
[8] 雍颹,钱锟,朱文昊,赵晓一,刘畅,潘洁. 成年恒牙牙髓切断后牙髓钙化的X线片评价[J]. 北京大学学报(医学版), 2023, 55(1): 88-93.
[9] 程昉,杨邵英,房星星,王璇,赵福涛. CCL28-CCR10通路在类风湿关节炎单核细胞迁移中的作用[J]. 北京大学学报(医学版), 2022, 54(6): 1074-1078.
[10] 王爽,彭楚芳,刘鹤. 新型生物陶瓷材料用于乳磨牙牙髓切断术的临床疗效[J]. 北京大学学报(医学版), 2022, 54(6): 1196-1201.
[11] 董尔丹. 心血管受体的信号转导与疾病[J]. 北京大学学报(医学版), 2022, 54(5): 796-802.
[12] 王跃,张爽,张虹,梁丽,徐玲,程元甲,段学宁,刘荫华,李挺. 激素受体阳性/人表皮生长因子受体2阴性乳腺癌临床病理特征及预后[J]. 北京大学学报(医学版), 2022, 54(5): 853-862.
[13] 程晓静,蒋栋,张连海,王江华,李雅真,翟佳慧,闫宝琪,张露露,谢兴旺,李子禹,季加孚. KRAS G12V特异性T细胞受体治疗恶性肿瘤的临床前研究[J]. 北京大学学报(医学版), 2022, 54(5): 884-895.
[14] 李芷晴,俞冰,蔡泽宇,王迎宝,张煦,周彪,方晓红,于芳,付毅,孙金鹏,李伟,孔炜. 柚皮素抑制马凡综合征小鼠胸主动脉瘤的形成[J]. 北京大学学报(医学版), 2022, 54(5): 896-906.
[15] 蔡天玉,朱振鹏,徐纯如,吉星,吕同德,郭振可,林健. 成纤维细胞生长因子受体2在肾透明细胞癌中的表达及意义[J]. 北京大学学报(医学版), 2022, 54(4): 628-635.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!