高尿酸对心肌细胞活力的影响及其相关机制探讨高尿酸对心肌细胞活力的影响及其相关机制探讨

摘要：目的　探讨高尿酸在体外对心肌细胞活力的影响及其相关机制。方法　实验分为空白对照组（Con组）、高尿酸组（HUA组）、抗氧化剂组（NAC组）、ERK抑制剂组（PD98059组）、P38抑制剂组（SB203580组）、高尿酸+抗氧化剂组（NAC+HUA组）、高尿酸+ERK抑制剂组（PD98059+HUA组）、高尿酸+P38抑制剂组（SB203580+HUA组）以及高尿酸+抗氧化剂+ERK抑制剂组（NAC+PD98059+HUA组），采用MTT法检测心肌细胞活力，免疫荧光染色及流式细胞仪检测细胞内活性氧（ROS）水平，Western blot检测细胞外信号调节激酶（ERK）及P38的蛋白表达水平。结果　经高尿酸处理后，心肌细胞活力在尿酸为0.15 g/L时下降最为明显。与Con组比较，HUA组的氧化压力明显上升；与HUA组比较，NAC+HUA组的氧化压力下降（P＜0.05）。与Con组相比，HUA组心肌细胞活力水平明显下降；与HUA组相比，NAC+HUA组、PD98059+HUA组及SB203580+HUA组的心肌细胞活力均上升（P＜0.05）。与Con组比较，HUA组ERK和P38的磷酸化水平明显增加；与HUA组相比，SB203580+HUA组的P38磷酸化水平明显下降而NAC+HUA组、PD98059+HUA组、NAC+PD98059+HUA组的ERK和P38的磷酸化水平均明显下降（P＜0.05）。结论　高尿酸在体外通过氧化应激激活ERK/P38信号通路抑制心肌细胞活力。

The effect of high uric acid on the activity of cardiomyocytes and its related mechanism

Abstract: Objective　To investigate the effect of high uric acid on cardiomyocyte activity in vitro and its related mechanism. Methods　H9C2 cells were divided into the control group (Con), the high uric acid group (HUA), the antioxidant group (NAC), the ERK inhibitor group (PD98059), the P38 inhibitor group (SB203580), the high uric acid + antioxidant group (NAC+HUA), the high uric acid + ERK inhibitor group (PD98059+HUA), the high uric acid + P38 inhibitor group (SB203580+HUA) and the high uric acid + antioxidant + ERK inhibitor group (NAC+PD98059+HUA). MTT method was used to detect the vitality of cardiomyocytes. Immunofluorescence staining and flow cytometry were used to detect the reactive oxygen species (ROS). Western blot assay was used to detect the protein expression levels of extracellular signal regulating kinase (ERK) and phosphorylated P38. Results　After treatment with high uric acid (0.15 g/L), the activity of cardiomyocytes decreased significantly. Compared with the Con group, the oxidative pressure increased significantly in the HUA group, while compared with the HUA group, the oxidative pressure decreased in the NAC+HUA group (P＜0.05). Compared with Con group, the activity level of cardiomyocytes decreased significantly in the HUA group, while compared with the HUA group, the activity levels of cardiomyocytes increased significantly in the NAC+HUA group, PD98059+HUA group and SB203580+HUA group (P＜0.05). Compared with the Con group, phosphorylation levels of ERK and P38 were increased significantly in HUA group. Compared with the HUA group, the phosphorylation levels of P38 of SB203580 group were decreased significantly, while the phosphorylation levels of ERK and P38 were decreased significantly in NAC+HUA group, PD98059+HUA group and NAC+PD98059+HUA group (P＜0.05). Conclusion　High uric acid inhibits myocardial cell activity by activating ERK/P38 signaling pathway through oxidative stress in vitro.