透明质酸对颞下颌关节滑膜细胞氧化应激的保护作用

Protective Effects of Hyaluronic Acid on Synovial Fibroblasts from Temporomandibular Joint Against Oxidative Stress Induced by Hydrogen Peroxide

目的:采用过氧化氢(H2O2)模拟颞下颌关节腔内的氧化应激状态,并利用系列浓度的高分子量透明质酸(hyaluronic acid,HA)进行干预,探讨氧化应激对滑膜细胞的影响及HA的干预作用。方法:实验动物为日本大耳白兔,通过外科手术获取颞下颌关节滑膜组织并进行细胞培养。采用系列浓度的H2O2进行氧化应激实验,探索合适的H2O2实验浓度,将H2O2与不同浓度的HA共同作用于培养细胞,观察细胞粘附情况,并采用噻唑蓝(methyl thiazol tetrazolium,MTT)法观察H2O2和不同浓度HA对滑膜细胞增殖的影响。结果:低浓度的H 2O 2(5、10μmol/L)对细胞活力没有抑制作用(P>0.05),而100μmol/L H2O2对细胞活力的抑制作用过强(P<0.05)。50μmol/L H2O2对细胞活力的抑制作用约为对照组的60%,因此选择此浓度为本实验的刺激浓度。中等浓度的HA(0.01%和0.05%)对滑膜细胞氧化应激的保护作用最强,与其他实验组比较差异有统计学意义(P<0.05);低浓度的HA效果不明显,与单纯H2O2组比较差异无统计学意义(P>0.05);而高浓度的HA抑制滑膜细胞增殖。结论:利用50μmol/L的H2O2可以模拟兔颞下颌关节滑膜细胞氧化应激状态,中等浓度的HA可以对滑膜细胞起到保护作用。

Objective:To observe the effects of oxygen free radicals on the synovial fibroblasts from the temporomandibular joint and the protective effects of hyaluronic acid against the oxidative stress.Methods:Synovial tissues were harvested from the rabbit temporomandibular joint and cultured en bloc.The synovial fibroblasts gained at 3-8 passages were used for further study.Hydrogen peroxide was introduced to mimic the effect of oxygen free radicals within the temporomandibular joint,and the hyaluronic acid was added to counteract the effects of reactive oxygen species.Results:There was no difference in the vitality of synovial fibroblasts between H2O2 groups(5μmol/L and 10μmol/L)and control group(P>0.05),while the 100μmol/L H2O2 overinhibited the viability of the synovial fibroblasts.50μmol/L H2O2 appropriately inhibited the activity of synovial fibroblasts(60%)and was selected as the ideal stimulus.In the serial concentrations of hyaluronic acid,the 0.01%and 0.05%groups significantly recovered the activity of synovial fibroblasts in contrast to the low or high concentrations of HA(P<0.05).Conclusion:Proper oxidative stress to the synovial fibroblasts can be induced by 50μmol/L H2O2,while the 0.01%and 0.05%HA can protect the synovial fibroblasts from the damage of oxygen free radicals.