瘦素对体外培养软骨细胞分泌NO和MMP-13的影响

目的:观察瘦素对体外培养兔关节软骨细胞分泌一氧化氮(NO)和基质金属蛋白酶-13(MMP-13)的影响。方法:获取2月龄兔原代软骨细胞,培养、鉴定、传代。将第3代软骨细胞按实验要求种板培养,饥饿12 h后,以不同浓度瘦素单独或与 TNF-&agr;联合干预48或96 h,测定各组细胞上清液中NO及MMP-13的浓度。结果:不同浓度瘦素(5,10,15和20 μg/mL)组NO浓度与空白对照组差异无统计学意义(P>0.05)；而不同浓度的瘦素与TNF-&agr;(10 ng/mL)联合应用后,与TNF-&agr;组差异有统计学意义(P<0.05)。空白对照组并未测出MMP-13的存在,不同瘦素浓度(10,50和100 ng/mL)组测得MMP-13浓度在剂量主效应和时间主效应均有统计学意义(P<0.05)。结论:瘦素在体外能协同TNF-&agr;促进兔关节软骨细胞分泌NO和MMP-13。

Effect of leptin on inducible NO and MMP-13 in rabbit articular chondrocytes in vitro

Objective To observe the in vitro effect of leptin, alone or in combination with tumor necrosis factor-alpha (TNF-α) on inducible nitric oxide (NO) and on inducible matrix metallo-proteinase-1 3 (MMP-13) in rabbit articular chondrocytes. Methods The chondrocytes from the articular cartilage of 2-month-old rabbits were cultivated and identified, and the second filial generation chondrocytes were cocultured on plates with different concentrations of leptin alone or in combination with TNF-α for 48 h or 96 h after 12 h starvation. The concentration of NO and MMP-13 was measured in the chondrocytes culture supernatant fluid. The results were statistically analyzed. Results There was no significant difference in the concentrations of NO between the different concentrations of leptin alone groups and the blank control group (P ＞ 0. 05). In combination with the same concentration of TNF-α (10 ng/mL), leptin could dose-dependently increase the concentration of NO in the chondrocytes culture supernatant fluid in vitro. There was significant value in average concentration of MMP-13 on the main effect of both time and dose (P ＜0. 05) . No MMP-13 was detected in the blank control group. Conclusion Leptin can induce MMP-13 and have synergistic induction effect on NO with TNF-α in rabbit articular chondrocytes in vitro.