摘要
目的:研究黄芪甲苷对人膝骨关节炎退变关节软骨VEGF表达的影响。方法:取人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VEGF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+25mmol/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+100mmol/L黄芪甲苷;F组:软骨细胞+500 mmol/L黄芪甲苷),取4 h、8 h、24 h、48 h、72 h采用实时荧光定量聚合酶链式反应(PCR)检测各组退变软骨内VEGF表达mRN A的表达情况。结果:培养的细胞为退变的软骨细胞;实时荧光定量PCR结果显示,不同浓度黄芪甲苷有上调退变膝关节软骨细胞VEGF的作用,且75 mmol/L黄芪甲苷在48 h时上调作用最好;在24 h、48 h、72 h不同浓度(50～100 mmol/l)黄芪甲苷膝关节软骨细胞VEGF表达量高于对照组(F=21.304,P<0.001)。结论:黄芪甲苷可通过调节退变软骨细胞合成VEGF延缓膝骨关节炎关节软骨细胞的退变。
Objective ： To investigate the effect of Astragaloside IV adjusting VEGF mRNA expression in chondrocyte of patients with knee osteoarthfitis. Methods：The cartilage of patients with knee osteoarthritis after operation was carried out and cell culture was made. When cells were more enough, and the third generation cells were given HE stain and fluoreseein stain with II - colla- gen to checkout whether they were the cartilage ceils. Then the third generation ceils were divided equally into six group（ group A： cartilage cells ＋ control group of DMEM ; group B ： cartilage cells ＋ concentration of AstragalosideIV was 25 mmol/L ; group C ： car- tilage cells ＋ concentration of AstragalosideIV was 50 mmol/L ; group D ： cartilage cells ＋ concentration of AstragalosideIV was 75 mmol/L;group E：cartilage cells ＋ concentration of AstragalosideIV was 100 mmol/L;group F：eartilage cells ＋ concentration of AstragalosideIV was 500 retool/L） ,at~ 4 h,8 h,24 h,48 h,72 h. Total RNA isolated from these samples was used for real - time PCR of VEGF. Results ： Cultured cells were cheekouted cartilage cells. Real - time PCR showed concentration of Astragaloside IV at 50 ~ 100 mmol/L upregulated mRNA expression of VEGF. Conclusion：Astragaloside IV can prevents cartilage degeneration of patients with knee osteoarthritis.
出处
《辽宁中医杂志》
CAS
2013年第5期1043-1045,共3页
Liaoning Journal of Traditional Chinese Medicine
基金
江苏省中医药管理局计划项目(HZ07089)
关键词
膝骨关节炎
退变
黄芪甲苷
软骨细胞
VEGF
knee osteoarthritis
degeneration
Astragaloside IV
cartilage cell
VEGF
