独活寄生汤含药血清对退变软骨细胞表达miRNA-140的影响

Effects of Serum Medicated with Duhuo Jisheng Tang(独活寄生汤) on the Expression of miRNA-140 in Degenerated Cartilages

目的:观察独活寄生汤含药血清对体外培养的大鼠退变软骨细胞表达miRNA-140的影响,探讨独活寄生汤防治骨关节炎的作用机制。方法:4周龄SD大鼠膝关节软骨建立体外培养的软骨细胞,采用Ⅱ型胶原原位杂交法对第3代(F3)软骨细胞进行鉴定。用10 ng·mL^(-1)白细胞介素-1β诱导F3软骨细胞,复制退变软骨细胞模型。成功后随机分为模型组和独活寄生汤含药血清组,分别在培养24,48,72 h后采用Taq Man real-time PCR法检测各组miRNA-140表达。结果:独活寄生汤含药血清组miRNA-140相对表达量随着干预时间的延长逐渐增加,差异有统计学意义(P<0.05)。结论:独活寄生汤治疗骨关节炎的部分作用机制是有效促进退变软骨细胞miRNA-140的表达。

Objective：To observe the effects of serum medicated with Duhuo Jisheng Tang （ 独活寄生汤） on the expression ofmiRNA-140 in degenerated cartilages cultured in vitro in order to explore the mechanism of Duhuo Jisheng Tang treating osteoarthritis.Methods：Chondrocytes were cultured in vitro in four-week old SD rats and the third generation （ F3 ） chondrocytes were identified by in situ hybridization of type Ⅱ collagen. F3 chondrocytes were induced by 10 ng ·mL-1 IL-1β,and the degenerated chondrocytes were duplicated.The rats were divided into a model group and a drug serum group.TaqMan real-time PCR method was used to detect the expression of miRNA-140 of each group after being cultured for 24,48 and 72 h respectively.Results：The expression time of miRNA-140 in serum medicated with Duhuo Jisheng Tang increased gradually with the prolonged time,and the difference was statistically significant （ P 〈 0.05 ） .Conclusion：The mechanism of Duhuo Jisheng Tang in the treatment of osteoarthritis is effective to promote the expression of miRNA-140 in degenerated cartilage.