中国骨质疏松杂志

小白菊内酯对地塞米松诱导的成骨细胞增殖和分化的影响

Effects of parthenolide on dexamethasone-induced osteoblast proliferation and differentiation

DOI：10.3969/j.issn.1006-7108.2023.09.010

中文关键词: 小白菊内酯成骨细胞 Wnt/β-catenin信号通路增殖成骨分化

英文关键词:parthenolide osteoblast Wnt/β-catenin signal pathway proliferation osteogenic differentiation

基金项目:医院培训振兴科研项目（201905）

作者	单位
张卓1 刘鹏1* 买若鹏1 苗磊1 陆钊罡2 董梦婷3	1.宁夏回族自治区人民医院脊柱与骨肿瘤外科，宁夏银川 750000 2.宁夏回族自治区人民医院临床药学室，宁夏银川 750000 3.宁夏回族自治区人民医质量管理办公室，宁夏银川 750000

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中文摘要:

目的探讨小白菊内酯（parthenolide，PTL）对地塞米松（dexamethasone，Dex）诱导的成骨细胞（MC3T3-E1）成骨分化及Wnt/β-catenin信号通路的影响。方法用浓度为2.5~80 μmol/L的小白菊内酯与地塞米松共同处理MC3T3-E1细胞，CCK-8法检测细胞增殖活性；将MC3T3-E1细胞分为对照组（Control组）、地塞米松组（Dex组）、小白菊内酯低、中、高浓度组（PTL-L组、PTL-M组、PTL-H组）、小白菊内酯高浓度+Wnt抑制剂组（PTL-H+DKK1组），碱性磷酸酶（ALP）染色检测细胞ALP活性，茜素红S染色观察细胞矿化结节形成；Western blot检测细胞Runt相关转录因子2（RUNX2）、骨钙素（OCN）、骨形成蛋白2（BMP2）、细胞周期蛋白Cyclin D1及Wnt/β-catenin通路相关蛋白表达水平。结果浓度为2.5~80 μmol/L的小白菊内酯可促进地塞米松诱导的MC3T3-E1细胞增殖，选择小白菊内酯浓度为5.0、10.0、20.0 μmol/L进行后续实验；与Control组相比，Dex组ALP活性显著降低，矿化结节减少，RUNX2、OCN、BMP2、Cyclin D1、Wnt3a、β-catenin蛋白表达水平显著降低（P<0.05）；与Dex组比较，PTL-L组、PTL-M组、PTL-H组ALP活性、矿化结节数依次升高，RUNX2、OCN、BMP2、Cyclin D1、Wnt3a、β-catenin蛋白表达水平依次升高（P<0.05）；与PTL-H组比较，PTL-H+DKK1组ALP活性与矿化结节显著降低，UNX2、OCN、BMP2、Cyclin D1、Wnt3a、β-catenin蛋白表达水平显著降低（P<0.05）。结论小白菊内酯通过激活Wnt/β-catenin信号通路，减轻地塞米松对成骨细胞增殖与分化的抑制作用。

英文摘要:

Objective To investigate the effects of parthenolide (PTL) on the differentiation of osteoblasts (MC3T3-E1) induced by dexamethasone (Dex) and the Wnt/β-catenin signal pathway. Methods MC3T3-E1 cells were treated with 2.5-80 μmol/L pyrethrone and dexamethasone. The cell proliferation activity was detected with CCK-8 method. MC3T3-E1 cells were divided into Control group, dexamethasone group (Dex group), low-, medium-, and high-concentration parthenolide groups (PTL-L group, PTL-M group, and PTL-H group), and high concentration parthenolide+Wnt inhibitor group (PTL-H+DKK1 group). ALP activity was detected with alkaline phosphatase (ALP) staining. The formation of mineralized nodules was observed with Alizarin red S staining. Western blotting was applied to detect the expression levels of Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), bone morphogenetic protein 2 (BMP2), cyclin D1, and Wnt/β-catenin pathway related proteins. Results Chrysanthemone with a concentration of 2.5-80 μmol/L promoted the proliferation of MC3T3-E1 cells induced by dexamethasone, so the Chrysanthemone with a concentration of 5.0, 10.0, and 20.0 μmol/L was selected for subsequent experiments. Compared with those in the control group, ALP activity and mineralized nodules in Dex group decreased obviously, and the expression levels of RUNX2, OCN, BMP2, Cyclin D1, Wnt3a, and β-catenin proteins decreased obviously (P<0.05). Compared with those in Dex group, ALP activity and the number of mineralized nodules in PTL-L group, PTL-M group, and PTL-H group increased in turn, and the protein expression levels of RUNX2, OCN, BMP2, Cyclin D1, Wnt3a, and β-catenin increased in turn (P<0.05). Compared with those in PTL-H group, ALP activity and mineralized nodules in PTL-H+DKK1 group decreased obviously, and the protein expression levels of UNX2, OCN, BMP2, Cyclin D1, Wnt3a, and β-catenin decreased obviously (P<0.05). Conclusion Parthenolide alleviates the inhibitory effects of dexamethasone on the proliferation and differentiation of osteoblasts by activating Wnt/β-catenin signal pathway.

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