[1]王刘玉,万全会,陈军.黄芪多糖对MC-3T3-E1成骨细胞增殖的影响及作用机制研究[J].中医正骨,2023,35(08):1-7.
　WANG Liuyu,WAN Quanhui,CHEN Jun.Effects and mechanism of Astragalan on proliferation of MC-3T3-E1 cells:an experimental study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2023,35(08):1-7.
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黄芪多糖对MC-3T3-E1成骨细胞增殖的影响及作用机制研究()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第35卷
期数:
2023年08期
页码:
1-7
栏目:
基础研究
出版日期:
2023-08-20

文章信息/Info

Title:
Effects and mechanism of Astragalan on proliferation of MC-3T3-E1 cells:an experimental study
作者:
王刘玉; 万全会; 陈军
南阳市第二人民医院,河南 南阳 473003
Author(s):
WANG Liuyu; WAN Quanhui; CHEN Jun
Nanyang Second General Hospital,Nanyang 473003,Henan,China
关键词:
骨质疏松;  成骨细胞;  细胞增殖;  黄芪多糖;  信号传导
Keywords:
osteoporosis;  osteoblasts;  cell proliferation;  astragalan;  signal transduction
摘要:
目的:探讨黄芪多糖对MC-3T3-E1成骨细胞增殖的影响及作用机制。方法:将培养的第3代MC-3T3-E1成骨细胞分为黄芪多糖低、中、高浓度组及黄芪多糖联合抑制剂干预组、对照组,黄芪多糖低、中、高浓度组分别用含有0.1 mol·L-1、1.0 mol·L-1、10 mol·L-1黄芪多糖的DMEM培养基进行培养,黄芪多糖联合抑制剂干预组用含有10 mol·L-1黄芪多糖和10 μmol·L-1 Compound C的DMEM培养基进行培养,对照组用DMEM培养基进行培养。干预24 h后,检测细胞增殖活力及碱性磷酸酶(alkaline phosphatase,ALP)、骨钙素(osteocalcin,OCN)、B淋巴细胞瘤-2(B-lymphoblastoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-related X protein,BAX)、半胱氨酸天冬氨酸蛋白酶(cysteine aspartic acid specific protease,Caspase)-3、磷酸化腺苷一磷酸活化蛋白激酶(phosphorylated AMP-activated protein kinase,p-AMPK)、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)等基因的蛋白表达情况。结果:①MC-3T3-E1成骨细胞增殖活力检测结果。黄芪多糖中、高浓度组细胞增殖活力均大于对照组(LSD-t=6.708,P=0.000; LSD-t=8.221,P=0.000)和黄芪多糖联合抑制剂干预组(LSD-t=2.314,P=0.049; LSD-t=5.286,P=0.001),黄芪多糖高浓度组细胞增殖活力大于黄芪多糖低、中浓度组(LSD-t=5.579,P=0.001; LSD-t=4.423,P=0.010)。②MC-3T3-E1成骨细胞成骨标志基因蛋白表达检测结果。黄芪多糖低、中、高浓度组细胞ALP、OCN的蛋白表达量均高于对照组(ALP:LSD-t=3.528,P=0.008; LSD-t=4.417,P=0.002; LSD-t=6.822,P=0.000; OCN:LSD-t=3.153,P=0.014; LSD-t=6.485,P=0.000; LSD-t=6.543,P=0.000)和黄芪多糖联合抑制剂干预组(ALP:LSD-t=2.414,P=0.042; LSD-t=3.155,P=0.013; LSD-t=5.892,P=0.000; OCN:LSD-t=2.339,P=0.047; LSD-t=5.926,P=0.000; LSD-t=14.546,P=0.000),黄芪多糖高浓度组细胞ALP、OCN的蛋白表达量高于黄芪多糖低、中浓度组(ALP:LSD-t=3.727,P=0.006; LSD-t=3.702,P=0.006; OCN:LSD-t=4.737,P=0.001; LSD-t=2.625,P=0.031)。③MC-3T3-E1成骨细胞线粒体凋亡途径相关基因蛋白表达检测结果。黄芪多糖中、高浓度组细胞Bcl-2的蛋白表达量均高于对照组(LSD-t=5.238,P=0.001; LSD-t=9.618,P=0.000)和黄芪多糖联合抑制剂干预组(LSD-t=3.821,P=0.006; LSD-t=8.246,P=0.000),黄芪多糖高浓度组细胞Bcl-2的蛋白表达量高于黄芪多糖低、中浓度组(LSD-t=8.875,P=0.001; LSD-t=6.102,P=0.000); 黄芪多糖中、高浓度组细胞BAX、Caspase-3的蛋白表达量均低于对照组(BAX:LSD-t=5.285,P=0.001; LSD-t=7.340,P=0.000; Caspase-3:LSD-t=3.654,P=0.006; LSD-t=6.875,P=0.000)和黄芪多糖联合抑制剂干预组(BAX:LSD-t=3.654,P=0.006; LSD-t=6.875,P=0.000; Caspase-3:LSD-t=2.940,P=0.019; LSD-t=6.314,P=0.000),黄芪多糖高浓度组细胞BAX、Caspase-3的蛋白表达量均低于黄芪多糖低、中浓度组(BAX:LSD-t=7.442,P=0.001; LSD-t=3.690,P=0.006; Caspase-3:LSD-t=4.496,P=0.002; LSD-t=4.642,P=0.002)。④MC-3T3-E1成骨细胞AMPK/eNOS信号通路相关基因蛋白表达检测结果。黄芪多糖低、中、高浓度组细胞p-AMPK、eNOS的蛋白表达量均高于对照组(p-AMPK:LSD-t=3.082,P=0.015; LSD-t=4.546,P=0.002; LSD-t=5.064,P=0.001; eNOS:LSD-t=3.395,P=0.009; LSD-t=5.873,P=0.000; LSD-t=7.327,P=0.000)和黄芪多糖联合抑制剂干预组(p-AMPK:LSD-t=5.819,P=0.000; LSD-t=6.731,P=0.000; LSD-t=6.961,P=0.000; eNOS:LSD-t=4.851,P=0.001; LSD-t=7.761,P=0.000; LSD-t=8.200,P=0.000)。黄芪多糖高浓度组细胞p-AMPK、eNOS的蛋白表达量高于黄芪多糖低浓度组(LSD-t=3.200,P=0.013; LSD-t=4.985,P=0.001)。结论:黄芪多糖能够促进MC-3T3-E1成骨细胞增殖,且该作用具有一定的浓度依赖性,其作用机制可能与调控线粒体凋亡途径及AMPK/eNOS信号通路有关。
Abstract:
Objective:To investigate the effects and mechanism of Astragalan on proliferation of MC-3T3-E1 cells.Methods:The MC-3T3-E1 cells were cultured,and the third-generation cells were collected and divided into Astragalan low-concentration group,Astragalan medium-concentration group,Astragalan high-concentratione group,Astragalan combined inhibitor intervention group and control group.The MC-3T3-E1 cells in control group were cultured in normal Dulbecco's Modified Eagle's Medium(DMEM),the ones in Astragalan low-,medium- and high-concentration group were cultured in DMEM supplemented with Astragalan with concentration of 0.1,1.0 and 10 mol/L respectively,and the ones in Astragalan combined inhibitor intervention group were cultured in DMEM supplemented with Astragalan and Compound C with concentration of 10 mol/L and 10 μmol/L respectively.After 24-hour culture,the MC-3T3-E1 cells proliferation activity and the protein expression levels of alkaline phosphatase(ALP),osteocalcin(OCN),B-lymphoblastoma-2(Bcl-2),Bcl-2-related X protein(BAX),cysteine aspartic acid specific protease(Caspase)-3,phosphorylated AMP-activated protein kinase(p-AMPK)and endothelial nitric oxide synthase(eNOS)were detected.Results:①The MC-3T3-E1 cells proliferation activity was higher in Astragalan middle- and high-concentration group compared to control group(LSD-t=6.708,P=0.000; LSD-t=8.221,P=0.000)and Astragalan combined inhibitor intervention group(LSD-t=2.314,P=0.049; LSD-t=5.286,P=0.001),and was higher in Astragalan high-concentration group compared to Astragalan low- and middle-concentration group(LSD-t=5.579,P=0.001; LSD-t=4.423,P=0.010).②The protein expression levels of ALP and OCN were higher in Astragalan low-,middle- and high-concentration group compared to control group(ALP:LSD-t=3.528,P=0.008; LSD-t=4.417,P=0.002; LSD-t=6.822,P=0.000; OCN:LSD-t=3.153,P=0.014; LSD-t=6.485,P=0.000; LSD-t=6.543,P=0.000)and Astragalan combined inhibitor intervention group(ALP:LSD-t=2.414,P=0.042; LSD-t=3.155,P=0.013; LSD-t=5.892,P=0.000; OCN:LSD-t=2.339,P=0.047; LSD-t=5.926,P=0.000; LSD-t=14.546,P=0.000),and was higher in Astragalan high-concentration group compared to Astragalan low- and middle-concentration group(ALP:LSD-t=3.727,P=0.006; LSD-t=3.702,P=0.006; OCN:LSD-t=4.737,P=0.001; LSD-t=2.625,P=0.031).③The protein expression level of Bcl-2 was higher in Astragalan middle- and high-concentration group compared to control group(LSD-t=5.238,P=0.001; LSD-t=9.618,P=0.000)and Astragalan combined inhibitor intervention group(LSD-t=3.821,P=0.006; LSD-t=8.246,P=0.000),and was higher in Astragalan high-concentration group compared to Astragalan low- and middle-concentration group(LSD-t=8.875,P=0.001; LSD-t=6.102,P=0.000).The protein expression levels of BAX and Caspase-3 were lower in Astragalan middle- and high-concentration group compared to control group(BAX:LSD-t=5.285,P=0.001; LSD-t=7.340,P=0.000; Caspase-3:LSD-t=3.654,P=0.006; LSD-t=6.875,P=0.000)and Astragalan combined inhibitor intervention group(BAX:LSD-t=3.654,P=0.006; LSD-t=6.875,P=0.000; Caspase-3:LSD-t=2.940,P=0.019; LSD-t=6.314,P=0.000),and was lower in Astragalan high-concentration group compared to Astragalan low- and middle-concentration group(BAX:LSD-t=7.442,P=0.001; LSD-t=3.690,P=0.006; Caspase-3:LSD-t=4.496,P=0.002; LSD-t=4.642,P=0.002).④The protein expression levels of p-AMPK and eNOS were higher in Astragalan low-,middle- and high-concentration group compared to control group(p-AMPK:LSD-t=3.082,P=0.015; LSD-t=4.546,P=0.002; LSD-t=5.064,P=0.001; eNOS:LSD-t=3.395,P=0.009; LSD-t=5.873,P=0.000; LSD-t=7.327,P=0.000)and Astragalan combined inhibitor intervention group(p-AMPK:LSD-t=5.819,P=0.000; LSD-t=6.731,P=0.000; LSD-t=6.961,P=0.000; eNOS:LSD-t=4.851,P=0.001; LSD-t=7.761,P=0.000; LSD-t=8.200,P=0.000),and was higher in Astragalan high-concentration group compared to Astragalan low-concentration group(LSD-t=3.200,P=0.013; LSD-t=4.985,P=0.001).Conclusion:Astragalan can promote the proliferation of MC-3T3-E1 cells,which exhibits a certain concentration-dependence,and its mechanism may be related to the regulation of mitochondrial-mediated apoptosis pathway and AMPK/eNOS signaling pathway.

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中医正骨2023年8月第35卷第8期 J Trad Chin Orthop Trauma,2023,Vol.35,No.8(总567)
(总568)中医正骨2023年8月第35卷第8期 J Trad Chin Orthop Trauma,2023,Vol.35,No.8
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更新日期/Last Update: 1900-01-01