本文關(guān)鍵詞： 雌激素 β雌二醇 生長(zhǎng)板 軟骨細(xì)胞 大鼠 C型利鈉肽 胰島素樣生長(zhǎng)因子 利鈉肽受體B　出處：《上海交通大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年08期 　論文類型：期刊論文

【摘要】：目的·探討不同濃度的17β雌二醇(17βE2)對(duì)大鼠生長(zhǎng)板軟骨細(xì)胞C型利鈉肽(CNP)、利鈉肽受體B(NPR-B)及胰島素樣生長(zhǎng)因子1(IGF1)的調(diào)控作用,及其對(duì)軟骨細(xì)胞增殖的影響。方法·Wistar大鼠8只,于出生后14 d處死,分離脛骨上段骺軟骨板,取得軟骨細(xì)胞并培養(yǎng)48 h后,于培養(yǎng)液中加入17βE2,并使其終濃度分別為10~(-4)、10~(-6)、10~(-8)、10~(-10)和10~(-12) mol/L,同時(shí)設(shè)空白對(duì)照組。繼續(xù)培養(yǎng)48 h。之后利用CCK8法測(cè)定軟骨細(xì)胞增殖能力,應(yīng)用ELISA法測(cè)定培養(yǎng)液中CNP和IGF1的濃度,并利用實(shí)時(shí)定量PCR檢測(cè)軟骨細(xì)胞CNP、NPR-B及IGF1 mRNA的相對(duì)水平。結(jié)果·不同濃度的17βE2對(duì)生長(zhǎng)板軟骨細(xì)胞增殖有顯著影響。當(dāng)17βE2的濃度為10~(-8) mol/L時(shí),其對(duì)軟骨細(xì)胞的促增殖效應(yīng)最強(qiáng);當(dāng)濃度進(jìn)一步增至10~(-4) mol/L時(shí),則抑制其增殖。隨著17βE2濃度的增加,細(xì)胞培養(yǎng)液中CNP的濃度以及軟骨細(xì)胞中CNP mRNA相對(duì)水平均出現(xiàn)明顯差異,以10~(-8) mol/L組CNP的濃度和mRNA水平最高,10~(-4) mol/L組為最低。IGF1同樣在10~(-8) mol/L組的濃度和mRNA表達(dá)水平達(dá)到最高,但最低值分別出現(xiàn)在10~(-10) mol/L組和10~(-12) mol/L組。軟骨細(xì)胞增殖水平與CNP的mRNA及蛋白濃度均呈正相關(guān)(均P=0.000),但與IGF1的mRNA及蛋白濃度水平無(wú)明顯相關(guān)性(均P0.05)。結(jié)論·17βE2對(duì)大鼠生長(zhǎng)板軟骨細(xì)胞增殖調(diào)控具有劑量-效應(yīng)關(guān)系,呈現(xiàn)出高濃度抑制、一定濃度范圍內(nèi)(10~(-10)~10~(-8) mol/L)促進(jìn)的作用;該作用與其調(diào)節(jié)生長(zhǎng)板軟骨細(xì)胞表達(dá)CNP水平呈正相關(guān)。
[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.
【作者單位】： 上海健康醫(yī)學(xué)院臨床醫(yī)學(xué)院;上海交通大學(xué)醫(yī)學(xué)院附屬瑞金醫(yī)院兒內(nèi)科;
【基金】：上海高校選拔培養(yǎng)優(yōu)秀青年教師科研專項(xiàng)基金(ZZjkxy13003) 金磊兒科中青年內(nèi)分泌醫(yī)師成長(zhǎng)科研基金(PEGRF201002008)~~
【分類號(hào)】：R965

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