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[1]王劉玉,萬(wàn)全會(huì),陳軍.黃芪多糖對(duì)MC-3T3-E1成骨細(xì)胞增殖的影響及作用機(jī)制研究[J].中醫(yī)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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黃芪多糖對(duì)MC-3T3-E1成骨細(xì)胞增殖的影響及作用機(jī)制研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第35卷
期數(shù):
2023年08期
頁(yè)碼:
1-7
欄目:
基礎(chǔ)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:
王劉玉; 萬(wàn)全會(huì); 陳軍
南陽(yáng)市第二人民醫(yī)院,河南 南陽(yáng) 473003
Author(s):
WANG Liuyu; WAN Quanhui; CHEN Jun
Nanyang Second General Hospital,Nanyang 473003,Henan,China
關(guān)鍵詞:
骨質(zhì)疏松;  成骨細(xì)胞;  細(xì)胞增殖;  黃芪多糖;  信號(hào)傳導(dǎo)
Keywords:
osteoporosis;  osteoblasts;  cell proliferation;  astragalan;  signal transduction
摘要:
目的:探討黃芪多糖對(duì)MC-3T3-E1成骨細(xì)胞增殖的影響及作用機(jī)制。方法:將培養(yǎng)的第3代MC-3T3-E1成骨細(xì)胞分為黃芪多糖低、中、高濃度組及黃芪多糖聯(lián)合抑制劑干預(yù)組、對(duì)照組,黃芪多糖低、中、高濃度組分別用含有0.1 mol·L-1、1.0 mol·L-1、10 mol·L-1黃芪多糖的DMEM培養(yǎng)基進(jìn)行培養(yǎng),黃芪多糖聯(lián)合抑制劑干預(yù)組用含有10 mol·L-1黃芪多糖和10 μmol·L-1 Compound C的DMEM培養(yǎng)基進(jìn)行培養(yǎng),對(duì)照組用DMEM培養(yǎng)基進(jìn)行培養(yǎng)。干預(yù)24 h后,檢測(cè)細(xì)胞增殖活力及堿性磷酸酶(alkaline phosphatase,ALP)、骨鈣素(osteocalcin,OCN)、B淋巴細(xì)胞瘤-2(B-lymphoblastoma-2,Bcl-2)、Bcl-2相關(guān)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)、內(nèi)皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)等基因的蛋白表達(dá)情況。結(jié)果:①M(fèi)C-3T3-E1成骨細(xì)胞增殖活力檢測(cè)結(jié)果。黃芪多糖中、高濃度組細(xì)胞增殖活力均大于對(duì)照組(LSD-t=6.708,P=0.000; LSD-t=8.221,P=0.000)和黃芪多糖聯(lián)合抑制劑干預(yù)組(LSD-t=2.314,P=0.049; LSD-t=5.286,P=0.001),黃芪多糖高濃度組細(xì)胞增殖活力大于黃芪多糖低、中濃度組(LSD-t=5.579,P=0.001; LSD-t=4.423,P=0.010)。②MC-3T3-E1成骨細(xì)胞成骨標(biāo)志基因蛋白表達(dá)檢測(cè)結(jié)果。黃芪多糖低、中、高濃度組細(xì)胞ALP、OCN的蛋白表達(dá)量均高于對(duì)照組(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)和黃芪多糖聯(lián)合抑制劑干預(yù)組(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),黃芪多糖高濃度組細(xì)胞ALP、OCN的蛋白表達(dá)量高于黃芪多糖低、中濃度組(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成骨細(xì)胞線粒體凋亡途徑相關(guān)基因蛋白表達(dá)檢測(cè)結(jié)果。黃芪多糖中、高濃度組細(xì)胞Bcl-2的蛋白表達(dá)量均高于對(duì)照組(LSD-t=5.238,P=0.001; LSD-t=9.618,P=0.000)和黃芪多糖聯(lián)合抑制劑干預(yù)組(LSD-t=3.821,P=0.006; LSD-t=8.246,P=0.000),黃芪多糖高濃度組細(xì)胞Bcl-2的蛋白表達(dá)量高于黃芪多糖低、中濃度組(LSD-t=8.875,P=0.001; LSD-t=6.102,P=0.000); 黃芪多糖中、高濃度組細(xì)胞BAX、Caspase-3的蛋白表達(dá)量均低于對(duì)照組(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)和黃芪多糖聯(lián)合抑制劑干預(yù)組(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),黃芪多糖高濃度組細(xì)胞BAX、Caspase-3的蛋白表達(dá)量均低于黃芪多糖低、中濃度組(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成骨細(xì)胞AMPK/eNOS信號(hào)通路相關(guān)基因蛋白表達(dá)檢測(cè)結(jié)果。黃芪多糖低、中、高濃度組細(xì)胞p-AMPK、eNOS的蛋白表達(dá)量均高于對(duì)照組(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)和黃芪多糖聯(lián)合抑制劑干預(yù)組(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)。黃芪多糖高濃度組細(xì)胞p-AMPK、eNOS的蛋白表達(dá)量高于黃芪多糖低濃度組(LSD-t=3.200,P=0.013; LSD-t=4.985,P=0.001)。結(jié)論:黃芪多糖能夠促進(jìn)MC-3T3-E1成骨細(xì)胞增殖,且該作用具有一定的濃度依賴性,其作用機(jī)制可能與調(diào)控線粒體凋亡途徑及AMPK/eNOS信號(hào)通路有關(guān)。
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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中醫(yī)正骨2023年8月第35卷第8期 J Trad Chin Orthop Trauma,2023,Vol.35,No.8(總567)
(總568)中醫(yī)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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通訊作者:陳軍 E-mail:[email protected]
更新日期/Last Update: 1900-01-01