上海交通大学学报(医学版) ›› 2017, Vol. 37 ›› Issue (8): 1074-.doi: 10.3969/j.issn.1674-8115.2017.08.005

• 论著(基础研究) • 上一篇    下一篇

雌二醇对大鼠生长板软骨细胞C 型利钠肽及胰岛素样生长因子1表达的影响

于波1,王俊祺2,王伟2,孙曼青2,肖园2   

  1. 1. 上海健康医学院 临床医学院,上海 201318;2. 上海交通大学 医学院附属瑞金医院儿内科,上海 200025
  • 出版日期:2017-08-28 发布日期:2017-09-28
  • 通讯作者: 肖园,电子信箱:yxiao.penicilinum@gmail.com
  • 作者简介:于波(1979—),女,讲师,硕士生;电子信箱:pipi_yb@126.com
  • 基金资助:
    上海高校选拔培养优秀青年教师科研专项基金(ZZjkxy13003);金磊儿科中青年内分泌医师成长科研基金(PEGRF201002008)

Effect of estrodial on C type natriuretic peptide and insulin like growth factor 1 expression in rat growth plate chondrocytes

YU Bo1, WANG Jun-qi2, WANG Wei2, SUN Man-qing2, XIAO Yuan2   

  1. 1. Shanghai University of Medicine & Health Sciences, School of Clinical Medicine, Shanghai 201318, China; 2. Pediatric Department, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2017-08-28 Published:2017-09-28
  • Supported by:
    Shanghai Municipal Scientific Research Special Fund for Outstanding University Youth Teacher, ZZjkxy13003;Jin Lei Pediatric Endocrinology Growth Research Fund for Young Physicians, PEGRF201002008

摘要:  目的 · 探讨不同浓度的 17β 雌二醇(17β E2)对大鼠生长板软骨细胞 C 型利钠肽(CNP)、利钠肽受体 B(NPR-B)及胰岛素样 生长因子 1(IGF1)的调控作用,及其对软骨细胞增殖的影响。方法 · Wistar 大鼠 8 只,于出生后 14 d 处死,分离胫骨上段骺软骨板, 取得软骨细胞并培养 48 h 后,于培养液中加入 17β E2,并使其终浓度分别为 10-4、10-6、10-8、10-10 和 10-12 mol/L,同时设空白对照组。 继续培养 48 h。之后利用 CCK8 法测定软骨细胞增殖能力,应用 ELISA 法测定培养液中 CNP 和 IGF1 的浓度,并利用实时定量 PCR 检 测软骨细胞 CNP、NPR-B 及 IGF1 mRNA 的相对水平。结果 · 不同浓度的 17β E2 对生长板软骨细胞增殖有显著影响。当 17β E2 的浓度 为 10-8 mol/L 时,其对软骨细胞的促增殖效应最强;当浓度进一步增至 10-4 mol/L 时,则抑制其增殖。随着 17β E2 浓度的增加,细胞培 养液中 CNP 的浓度以及软骨细胞中 CNP mRNA 相对水平均出现明显差异,以 10-8 mol/L 组 CNP 的浓度和 mRNA 水平最高,10-4 mol/L 组为最低。IGF1 同样在 10-8 mol/L 组的浓度和 mRNA 表达水平达到最高,但最低值分别出现在 10-10 mol/L 组和 10-12 mol/L 组。软骨细 胞增殖水平与 CNP 的 mRNA 及蛋白浓度均呈正相关(均 P=0.000),但与 IGF1 的 mRNA 及蛋白浓度水平无明显相关性(均 P>0.05)。 结论 · 17β E2 对大鼠生长板软骨细胞增殖调控具有剂量- 效应关系,呈现出高浓度抑制、一定浓度范围内(10-10 ~ 10-8 mol/L)
 促进的作用;该作用与其调节生长板软骨细胞表达 CNP 水平呈正相关。

关键词: 雌激素, 17&beta, 雌二醇, 生长板, 软骨细胞, 大鼠, C 型利钠肽, 胰岛素样生长因子1, 利钠肽受体B

Abstract:  Objective · To observe effect of 17β estrodial (17β E2) with different concentrations on C type natriuretic peptide (CNP), insulin like growth factor 1 (IGF1), and natriuretic peptide receptor B (NPR-B) expression and proliferation of growth plate chondrocytes of rats in vitro.  Methods · Eight Wistar rats were sacrificed and their epiphyseal cartilages of the upper tibias were separated to obtain chondrocytes on the 14th day after birth. Then chondrocytes were cultured with 17β E2 in different concentrations (10-4、10-6、10-8、10-10 and 10-12 mol/L) for 48 h, while control group was cultured without 17β E2. CCK8 method, ELISA and qRT-PCR were used to analyze the proliferation of chondrocytes, the levels of CNP and IGF1 in culture medium and mRNA levels of CNP, NPR-B and IGF1, respectively.  Results · 17β E2 in different concentrations affected the proliferation of growth plate chondrocytes significantly. When the concentration of 17β E2 was 10-8 mol/L, it had the strongest effect on the cell proliferation. When the concentration increased to 10-4 mol/L, the proliferation of chondrocytes was inhibited. With the increasement of 17β E2 concentration, the levels of CNP in the culture medium and the mRNA levels of CNP in the chondrocytes were significantly different. The highest levels of CNP protein and mRNA both appeared in 10-8 mol/L group, while the lowest levels both appeared in 10-4 mol/L group. IGF1 and its mRNA also reached the highest levels in 10-8 mol/L group, but the lowest concentration and mRNA level were in 10-10 mol/L group and 10-12 mol/L, respectively. Both CNP mRNA and protein levels were positive correlated with the proliferation of chondrocytes (P=0.000). Nevertheless, there was no significant correlation between the proliferation of chondrocytes and IGF1 mRNA or protein levels (P>0.05).  Conclusion · 17β E2 modulates proliferation of rat growth plate chondrocytes in a dose-effect manner. It enhances proliferation at relatively low concentrations (10-10-10-8 mol/L) and inhibits proliferation at high concentration. This effect is positively related to CNP expression in chondrocytes.

Key words: 17&beta, estrodial, growth plate, chondrocyte, rat, C type natriuretic peptide, insulin like growth factor 1, natriuretic peptide receptor B