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

发布时间：2018-02-28 12:03

本文关键词： 雌激素 β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利钠肽受体B　出处：《上海交通大学学报(医学版)》2017年08期 　论文类型：期刊论文

【摘要】：目的·探讨不同浓度的17β雌二醇(17βE2)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17βE2,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17βE2对生长板软骨细胞增殖有显著影响。当17βE2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17βE2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P0.05)。结论·17βE2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。
[Abstract]:Objective: to investigate the effects of 17 尾 estradiol 17 尾 E2 (17 尾 E 2) on the growth plate chondrocytes C-type natriuretic peptide (CNP), natriuretic peptide receptor (BnPR-B1) and insulin-like growth factor 1IGF1 (IGF1) in chondrocytes of rats. Methods: the proliferation of chondrocytes was observed in 8 Wistar rats. The epiphyseal chondrocytes were isolated from the epiphyseal plate of the upper tibia on the 14th day after birth, and the chondrocytes were obtained and cultured for 48 h. 17 尾 E _ 2 was added to the culture medium and the final concentrations were 10 ~ (10) ~ (-4) ~ 10 ~ (-6) ~ 10 ~ (-10) ~ 10 ~ (-10) mol 路L ~ (-1) and 10 ~ 10 ~ (-12) mol 路L ~ (-1), respectively, and a blank control group was set up. The proliferation of chondrocytes was determined by CCK8 assay and the concentration of CNP and IGF1 in culture medium was determined by ELISA method. The relative levels of CNPnPR-B and IGF1 mRNA in chondrocytes were measured by real-time quantitative PCR. Results: the proliferation of chondrocytes was significantly affected by 17 尾 E2 at different concentrations. When the concentration of 17 尾 E2 was 10 ~ 8 mol/L, the effect of 17 尾 E _ 2 on the proliferation of chondrocytes was the strongest. With the increase of 17 尾 E 2 concentration, the concentration of CNP in cell culture medium and the relative level of CNP mRNA in chondrocytes were significantly different. The highest concentration of CNP and the highest level of mRNA were found in the mol/L group. The lowest. IGF1 level and mRNA expression level in the mol/L group were also the highest in the mol/L group, and the expression of mRNA was the highest in the mol/L group, and the expression of mRNA was the highest in the mol/L group, and the expression of mRNA was the highest in the mol/L group. However, the lowest values were found in the mol/L group and the mol/L group, respectively. There was a positive correlation between the proliferation of chondrocytes and the mRNA and protein concentrations of CNP (all P < 0.000), but no significant correlation with the mRNA and protein concentrations of IGF1 (P 0.05). Conclusion\\\. The proliferation regulation of long plate chondrocytes has a dose-effect relationship. The inhibitory effect of high concentration and the effect of 10 ~ 10 ~ 10 ~ (-8) mol 路L ~ (-1) 路L ~ (-1) on the expression of CNP in chondrocytes of growth plate were positively correlated.
【作者单位】：上海健康医学院临床医学院;上海交通大学医学院附属瑞金医院儿内科;
【基金】：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)~~
【分类号】：R965

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