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黄芪甲苷对人膝骨关节炎退变关节软骨IL-1β表达的影响

汤晓晨 俞峰 孙书龙 姜宏 龚正丰 许建安 王拥军

汤晓晨, 俞峰, 孙书龙, 姜宏, 龚正丰, 许建安, 王拥军. 黄芪甲苷对人膝骨关节炎退变关节软骨IL-1β表达的影响[J]. 南京中医药大学学报, 2013, 29(1): 48-52.
引用本文: 汤晓晨, 俞峰, 孙书龙, 姜宏, 龚正丰, 许建安, 王拥军. 黄芪甲苷对人膝骨关节炎退变关节软骨IL-1β表达的影响[J]. 南京中医药大学学报, 2013, 29(1): 48-52.
TANGXiao-chen, YUFeng, SUNShu-long, JIANGHong, GONGZheng-feng, XUJian-an, WANGYong-jun. Effect of Astragaloside IV on IL-1β Expression of Degenerative Joint Cartilage in Human Knee Osteoarthritis[J]. Journal of Nanjing University of traditional Chinese Medicine, 2013, 29(1): 48-52.
Citation: TANGXiao-chen, YUFeng, SUNShu-long, JIANGHong, GONGZheng-feng, XUJian-an, WANGYong-jun. Effect of Astragaloside IV on IL-1β Expression of Degenerative Joint Cartilage in Human Knee Osteoarthritis[J]. Journal of Nanjing University of traditional Chinese Medicine, 2013, 29(1): 48-52.

黄芪甲苷对人膝骨关节炎退变关节软骨IL-1β表达的影响

Effect of Astragaloside IV on IL-1β Expression of Degenerative Joint Cartilage in Human Knee Osteoarthritis

  • 摘要: 目的 研究黄芪甲苷对人膝骨关节炎退变关节软骨IL-1β表达的影响。方法 取膝骨关节炎患者手术后的退变膝关节软骨,进行细胞培养,将培养的软骨细胞分离传代后,取第三代细胞,通过HE、Ⅱ型胶原免疫荧光染色进行软骨细胞鉴定,并进行IL-1β免疫荧光染色。观察软骨细胞退变情况后分为6组(A组:软骨细胞+DMEM对照组;B组:软骨细胞+25 mmol/L黄芪甲苷;C组:软骨细胞+50 mmol/L黄芪甲苷;D组:软骨细胞+75 mmol/L黄芪甲苷;E组:软骨细胞+100 mmol/L黄芪甲苷;F组:软骨细胞+500 mmol/L黄芪甲苷),取4、8、24、48、72 h采用实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测各组退变软骨细胞内IL-1β mRNA的表达情况。结果 ①形态观察。原代细胞中梭形细胞和多角形细胞并存,细胞核偏于细胞的一侧而贴近细胞膜,细胞纤丝较长。HE染色细胞质为红色,胞膜呈蓝色;甲苯胺蓝染色细胞质和胞膜均呈深蓝色;Ⅱ型胶原染色阳性为绿色荧光,细胞核采用4’,6-二脒基-2-苯基吲哚复染,在不同波段荧光激发下,胞浆和胞膜可见清晰绿色荧光,细胞核区域未见明显绿色荧光,证明培养细胞表达特征性基因Ⅱ型胶原,主要分布在胞浆和细胞膜上,证明所培养的细胞为软骨细胞。IL-1β免疫荧光染色可见清晰绿色荧光,软骨细胞以多角形多见。②IL-1β mRNA表达。不同时间点间软骨细胞IL-1β mRNA表达水平有差异(F=13.236,P<0.01);不同组间软骨细胞IL-1β mRNA表达水平有差异(F=98.762,P<0.01),A组高于B组、C组、D组、E组(P<0.05~0.01),F组高于A组、B组、C组、D组、E组(P<0.05~0.01);时间因素和组间因素存在交互效应(F=7.262,P<0.01)。结论 一定浓度的黄芪甲苷能通过抑制退变软骨细胞合成IL-1β,延缓软骨细胞退变;浓度过高则会促进退变软骨细胞合成IL-1β,加速软骨细胞退变。

     

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出版历程
  • 收稿日期:  2012-10-19
  • 修回日期:  2012-11-28
  • 刊出日期:  2013-01-10

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