黄芪多糖对激素性股骨头坏死模型骨细胞凋亡和SP1/MEK/ERK轴的影响  

Effect of astragalus polysaccharides on osteocyte apoptosis and SP1/MEK/ERK axis in cell models of steroid-induced osteonecrosis of femoral head

作　　者：卢敏[1] 许晓彤 申楠 张申尧[2] Lu Min;Xu Xiao-tong;Shen Nan;Zhang Shen-yao(Department of Orthopedics,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410021,China;Department of Orthopedics,The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410005,China)

机构地区：[1]湖南中医药大学第一附属医院骨伤科,湖南长沙410021 [2]湖南中医药大学第二附属医院骨伤科,湖南长沙410005

出　　处：《中国现代医学杂志》2023年第12期33-40,共8页China Journal of Modern Medicine

基　　金：湖南省自然科学基金(No:2022JJ40328)。

摘　　要：目的探究黄芪多糖对激素性股骨头坏死(SONFH)模型骨细胞凋亡和SP1/MEK/ERK轴的影响。方法通过地塞米松诱导小鼠骨细胞(MLO-Y4)建立SONFH细胞模型,加入300μg/mL黄芪多糖处理后通过MTT和流式细胞术检测骨细胞活性和细胞凋亡。进一步在细胞中过表达或者敲降SP1,通过Western blotting验证SP1表达对MEK/ERK通路活性的影响。结果SONFH模型组48 h后光密度值较对照组降低(P<0.05),SONFH+黄芪多糖组较SONFH+PBS组升高(P<0.05)。SONFH模型组细胞凋亡率较对照组高(P<0.05),SONFH+黄芪多糖组较SONFH模型组低(P<0.05),SONFH模型组Bax、Cleaved-caspase-3蛋白相对表达量较对照组高(P<0.05),Bcl-2蛋白相对表达量较对照组低(P<0.05),SONFH+黄芪多糖组Bax、Cleavedcaspase-3蛋白相对表达量较SONFH模型组低(P<0.05),Bcl-2蛋白相对表达量较SONFH模型组高(P<0.05)。SONFH模型组SP1 mRNA、蛋白相对表达量较对照组高(P<0.05),SONFH+黄芪多糖组较SONFH模型组低(P<0.05)。SONFH+黄芪多糖组SP1 mRNA相对表达量较SONFH模型组低,SONFH+黄芪多糖+OE-SP1组较SONFH+黄芪多糖+OE-NC组高。SONFH模型组细胞活性较对照组降低(P<0.05),SONFH+黄芪多糖组较SONFH模型组升高(P<0.05),SONFH+黄芪多糖组与SONFH+黄芪多糖+OE-NC组比较,差异无统计学意义(P>0.05),SONFH+黄芪多糖+OE-SP1组较SONFH+黄芪多糖组降低(P<0.05)。SONFH模型组细胞凋亡率和Bax、Cleaved-caspase-3蛋白相对表达量较对照组高(P<0.05),SONFH+黄芪多糖组较SONFH模型组低(P<0.05),SONFH+黄芪多糖+OE-SP1组较SONFH+黄芪多糖+OE-NC组高(P<0.05);SONFH模型组Bcl-2蛋白相对表达量较对照组低(P<0.05),SONFH+黄芪多糖组较SONFH模型组低(P<0.05),SONFH+黄芪多糖+OE-SP1组较SONFH+黄芪多糖+OE-NC组低(P<0.05)。OE-SP1组SP1 mRNA相对表达量较OENC组高,sh-SP1组较sh-NC组低。OE-SP1组SP1蛋白相对表达量较对照组、OE-NC组高(P<0.05),shSP1组较sh-NC低(P<0.05);OE-SP1组MEK、p-ERK蛋白相对表达量较对�Objective To explore the effect of astragalus polysaccharides on osteocyte apoptosis and SP1/MEK/ERK axis in cell models of steroid-induced osteonecrosis of femoral head(SONFH).Methods The cell model of SONFH was established by treating mouse osteocytes(MLO-Y4)with dexamethasone.After modeling,the SONFH cells were treated with 300μg/mL of astragalus polysaccharides,followed by the MTT assay and flow cytometry to detect the cell viability and apoptosis.Furthermore,SP1 was overexpressed or knocked down in SONFH cells,and the effect of SP1 expression on the activity of the MEK/ERK pathway was verified by Western blotting.Results The 48-hour optical density of the SONFH model group was lower than that of the control group(P<0.05),while that in the SONFH+astragalus polysaccharides group was higher compared with the SONFH+PBS group(P<0.05).The cell apoptosis rate in the SONFH model group was higher than that in the control group(P<0.05),while that in the SONFH+astragalus polysaccharides group was lower compared with the SONFH model group(P<0.05).Compared with the control group,the protein expressions of Bax and cleaved caspase-3 were higher,but the protein expression of Bcl-2 was lower in the SONFH model group(P<0.05).Compared with the SONFH model group,the protein expressions of Bax and cleaved caspase-3 were lower,but the protein expression of Bcl-2 was higher in the SONFH+astragalus polysaccharides group(P<0.05).The mRNA and protein expressions of SP1 in the SONFH model group were higher than those in the control group(P<0.05),whereas those in the SONFH+astragalus polysaccharides group were lower compared with the SONFH model group(P<0.05).The mRNA expression of SP1 in the SONFH+astragalus polysaccharides group was lower than that in the SONFH model group,while that in the SONFH+astragalus polysaccharides+OE-SP1 group was higher compared with the SONFH+astragalus polysaccharides+OE-NC group(P<0.05).The cell viability in the SONFH model group was lower than that in the control group(P<0.05),while that in the SONFH+a

关 键 词：激素性股骨头坏死 黄芪多糖 SP1 细胞凋亡 

分 类 号：R681.6[医药卫生—骨科学]