干扰小RNA下调瘦素受体抑制人骨关节炎软骨细胞白细胞介素-1β和一氧化氮的信使RNA表达

Down-regulated Leptin Receptor by Small Interfering RNA Inhibits the Messenger RNA Expressions of Interleukin-1β and Nitric Oxide of Human Osteoarthritis Chondrocytes

目的观察干扰小RNA(si RNA)下调瘦素受体对人骨关节炎软骨细胞白细胞介素(IL)-1β和一氧化氮(NO)表达的抑制作用,为临床基础研究提供参考依据。方法取全膝置换术患者残存的无菌膝关节软骨进行分离培养人关节软骨细胞,取第3代软骨细胞对瘦素受体的Ob-Rb的分别用携带si RNA的脂质体(实验组)和空白脂质体(对照组)进行转染,转染成功后24、48、72 h采用荧光定量聚合酶链反应测前炎性因子基因IL-1β和NO的信使RNA表达量。结果转染成功后24、48及72 h,实验组IL-1β的表达逐渐略有上升,NO的m RNA表达逐渐下降,但均明显低于对照组(P<0.05),且随时间推移差值逐渐变大。结论 si RNA技术下调瘦素受体可明显抑制人骨关节炎软骨细胞IL-1β和NO的m RNA的表达水平。

Objective To study the effect of down-regulated leptin receptor by small interfering RNA （siRNA） in inhibiting the messenger RNA （mRNA） expressions of interleukin （IL）-1β and nitric oxide （NO） of human osteoarthritis chondrocytes, in order to provide reference for basic clinical research. Methods Cartilage was harvested under sterile conditions from osteoarthritis knee joints in patients undergoing total knee arthroplasty. Human articular chondrocytes were isolated and the cells were cultured in vitro. The cells in the 3rd passage were transferred by siRNA Ob-Rb （experimental group） and blank Ob-Rb （control group）, respectively. Then mRNA expressions of IL-1β and NO were tested by quantitative polyrnerase chain reaction at hour 24, 48 and 72 after successful transfection. Results The mRNA expressions of IL-1β increased slightly and that of NO declined slightly at hour 24, 48 and 72 after transfection in the treatment group, but they all were significantly lower than those in the control group （P 〈 0.05）, and the differences became much larger as time went on. Conclusion Leptin receptor under siRNA technology can significantly inhibit the mRNA expressions of IL-1β and NO in human osteoarthritis chondrocytes,