世界中医药
文章摘要
中药淫羊藿苷对TDP-43介导的骨关节炎软骨细胞病变的作用机制
Action Mechanism Analysis of Icariin on TDP-43 Mediated Chondrocytic Lesions in Osteoarthritis
投稿时间:2018-08-17  
DOI:10.3969/j.issn.1673-7202.2019.07.010
中文关键词: 骨关节炎  软骨细胞病变  淫羊藿苷  TDP-43  作用机制  中药  慢病毒转染  细胞凋亡
英文关键词: Osteoarthritis  Chondrocyte lesions  Icariin  TDP-43  Mechanism of action  Traditional Chinese medicine  Lentivirus transfection  Apoptosis
基金项目:大连市卫生和计划生育委员会医学科学研究计划项目([2016]111号)
作者单位
孙海峰,杨茂伟,时永臣,王守赟 大连市第二人民医院创伤外科大连116011 
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中文摘要:
      目的:研究分析中药淫羊藿苷对TDP-43介导的骨关节炎软骨细胞病变的作用机制,从而为临床有效治疗骨关节炎软骨细胞病变提供参考依据。方法:选取并构建TDP-43慢病毒转染人软骨细胞,并予以不同浓度的中药淫羊藿苷进行干预。采用荧光定量PCR检测不同组别人软骨细胞的TDP-43基因表达情况,采用流式细胞仪检测TDP-43慢病毒转染的人软骨细胞加入中药淫羊藿苷前后细胞凋亡情况,通过连续细胞计数观察细胞增殖情况。分别比较3组人软骨细胞中靶基因TDP-43与Ⅰ型胶原蛋白α1表达情况,TDP-43慢病毒转染的人软骨细胞加入中药淫羊藿苷前后细胞凋亡情况,加入不同浓度中药淫羊藿苷培养后的软骨细胞计数水平。结果:TDP-43慢病毒转染的人软骨细胞中TDP-43基因表达水平为(16.02±1.45),比正常人软骨细胞、空白慢病毒转染的人软骨细胞的(1.86±0.13)、(1.88±0.12)高,差异有统计学意义(P<0.05);而3组Ⅰ型胶原蛋白α1基因表达差异无统计学意义(P>0.05)。加入不同浓度的中药淫羊藿苷后TDP-43慢病毒转染的人软骨细胞凋亡率相比加入中药淫羊藿苷前均降低,差异有统计学意义(P<0.05)。培养3 d后、6 d后、12 d后高浓度中药淫羊藿苷培养的软骨细胞计数比中浓度高,而中浓度中药淫羊藿苷培养的软骨细胞计数水平比低浓度高,差异有统计学意义(P<0.05)。结论:TDP-43可能在骨关节炎软骨细胞病变过程中发挥负调控作用,而中药淫羊藿苷可通过改善上述过程,改善TDP-43介导的骨关节炎软骨细胞病变。
英文摘要:
      To study and analyze the mechanism of icariin on TDP-43 mediated chondrocytic lesions in osteoarthritis,so as to provide a reference for effective treatment of chondrocystic lesions in osteoarthritis.Methods:TDP43 lentivirus was transfected into human chondrocytes,and intervened by icariin with different concentrations.Fluorescence quantitative PCR was used to detect the expression of TDP-43 gene in different groups of human chondrocytes.Flow cytometry was used to detect the apoptosis of human chondrocytes transfected with TDP43 lentivirus before and after the addition of icariin.Three sets of target genes in human cartilage cells TDP43 and collagen expression in alpha 1 Ⅰ type,the cell apoptosis of TDP43 lentivirus transfection human cartilage cells before and after adding icariin,cartilage cell count after cultivation by adding different concentration of icariin were compared respectively.Results:TDP43 gene expression level in human chondrocytes transfected with TDP43(16.02±1.45)was higher than that in normal human chondrocytes and human chondrocytes transfected with lentivirus(1.86±0.13)and(1.88±0.12),with statistically significant differences(all P<0.05).The comparison of Ⅰ type collagen and three groups of alpha 1 gene expression levels were not obvious(P>0.05).The apoptosis rate of human chondrocytes transfected with TDP43 lentivirus after adding different concentrations of icariin was lower than before adding icariin,with statistically significant differences(all P<0.05).The chondrocytes cultured with high concentration of icariin after 3 d,6 d and 12 d cultivation were higher than the medium concentration,while the chondrocytes cultured with medium concentration of icariin were higher than the low concentration.And one-way analysis of variance showed that the difference between the groups was statistically significant(all P<0.05).Conclusion:TDP-43 may play a negative regulatory role in osteoarthritis chondrocytic lesions,while icariin can improve the above process and further improve the TDP-43 mediated chondrocytic lesions in osteoarthritis.
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