心房颤动患者血清TNF-α、TGF-β水平及其对心肌成纤维细胞趋化作用的影响

Serum Levels of Tumor Necrosis Factor-alpha and Transforming Growth Factor-beta in Atrial Fibrillation and Its Chemotatic Effects on Cardiac Fubroblast

目的研究心房颤动与非心房颤动患者血清中转化生长因子β(TGF-β)、肿瘤坏死因子α(TNF-α)的动态变化及其对心肌成纤维细胞趋化运动诱导能力的差异。方法采用酶联免疫吸附法检测心房颤动与非心房颤动患者血清中TGF-β、TNF-α蛋白含量;培养新生SD大鼠的心肌成纤维细胞,取3~4代细胞采用Transwell装置检测不同浓度(1μg/L、10μg/L、50μg/L)的TGF-β、TNF-α诱导成纤维细胞运动的能力差异。结果血清中TGF-β、TNF-α含量各组均有明显差异,其中持续性心房颤动组最高,非心房颤动房性心律失常组明显高于阵发性心房颤动组,对照组最低;在不同浓度(1μg/L、10μg/L、50μg/L)TGF-β、TNF-α的趋化诱导作用下,迁移至下室的细胞50μg/L组较其它浓度组明显增高,10μg/L组次之,与其它各组比较均具有统计学意义。Logistic回归分析表明,迁移至滤膜下的细胞数与血清中TGF-β、TNF-α含量有相关性。结论各组血清中TGF-β、TNF-α含量不同,说明心房颤动的发生与潜在的炎症状态有关;TGF-β、TNF-α对心肌成纤维细胞具有趋化作用,心房颤动患者心肌成纤维细胞的趋化作用部分是由TNF-α、TGF-β介导的;血清中TNF-α、TGF-β水平变化间接反映了心肌损伤的存在、范围和程度。

Aim To investigate the different serum levels of tumor necrosis factor-alpha( TNF-α) and transforming grouth factor-beta( TGF-β) between atrial fibrilliation and other control groups,and its chemotatic effects on cardiac fubroblast. Methods Serum concentrations of TNF-α and TGF-β were measured by enzyme-linked immune-absorbent assay( ELISA); Cardiac fibroblast were isolated from the venticles of 1 ~ 3 days Sprague-Dawley neonatal rats and passaged three to four generation,the transwell chamber assay were used to test the different chemotatic effects of the cardiac fibroblasts with the different concentration( 1 μg / L,10 μg / L,50 μg / L) of TNF-α and TGF-β. Results Compared with control group,the serum of TNF-α and TGF-β in other groups were obviously increased. The group of persistent atrial fibrillation( per-Af) was the highest,the serum of TNF-α and TGF-β in non-Af atrial arrhythmia( AA) was higher than paroxysmal Af( per-Af),the control group was the least. In different concentration( 1 μg / L,10 μg / L,50 μg / L) of TNF-αand TGF-β chemotaxis induced,the number of cells that migrated through the polycarbonate membrane were different,50μg /L group were significantly increased compared with other concentrations,the number of migrated cell in 10 μg /L group was higher than 1 μg / L group. The multiple Logistic regression analysis showed that the migrated cells in the lower surface were related to the concentration of TNF-α and TGF-β. Conclusions The different concentration of TNF-α and TGF-β among each group show that the occurrence of atrial fibrillation is associated with the potential inflammatory state.TNF-α and TGF-β have chemotaxis effect on cardiac fibroblast,its level is correlated with chemotatic movement. The chemotaxis of the serum with atrial fibrillation patients is partly mediated by TNF-α and TGF-β and it plays an important role in atrial fibrosis. The concentration of TNF-α and TGF-β can be used to reflect indirectly the presence,severity,extent of the myocardial damage.