姜黄素通过抑制NF-κB信号通路减轻高糖致H9C2心肌细胞损伤的实验研究

Circumin attenuates high glucose-induced H9C2 cardiomyocyte injury via NF-κB signaling pathway

关键词:  姜黄素  心肌细胞  炎症反应  氧化应激  NF-κB

Key words:  curcumin  cardiomyocytes  inflammation  oxidative stress  NF-κB

摘要:

[目的] 观察姜黄素对炎症反应、氧化应激相关指标及细胞核转录因子κB（NFκB）信号通路的影响，探讨其减轻高糖致H9C2细胞损伤的作用机制。[方法] 高糖孵育建立H9C2心肌细胞损伤模型。实验分为正常对照组、高糖对照组和姜黄素各剂量组（2.5、5、10、20 μmol/L）。酶联免疫吸附（ELISA）法测定细胞培养液上清中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、乳酸脱氢酶（LDH）的含量以及细胞内丙二醛（MDA）、超氧化物歧化酶（SOD）的含量。蛋白质印迹（Western Blot）法检测细胞内IκB激酶β（IKKβ）、NF-κB p65、p-NF-κB p65的蛋白表达水平。[结果] 与正常对照组比较，高糖对照组H9C2心肌细胞活性降低，细胞培养液上清中TNF-α、IL-6、LDH含量明显增加（P<0.05），心肌细胞内MAD含量增加（P<0.05）、SOD含量降低（P<0.05），心肌细胞内IKKβ蛋白表达增加（P<0.05），NFκB p65蛋白表达变化不明显，但p-NF-κB p65的蛋白表达水平增加（P<0.05）；与高糖对照组比较，姜黄素各剂量组预处理6 h可呈剂量依赖性增强H9C2细胞活性（P<0.05），降低细胞培养液上清中TNF-α、IL-6、LDH含量（P<0.05），降低细胞内MDA含量、增加SOD含量（P<0.05），降低IKKβ蛋白表达（P<0.05），降低p-NF-κBp65蛋白表达（P<0.05）。[结论] 姜黄素可通过抑制NF-κB信号通路过度激活，降低炎症反应及氧化应激状态，减轻高糖诱导的H9C2心肌细胞损伤。

Abstract:

[Objective] To observe the effects of curcumin on inflammation,oxidative and NF-κB signaling pathway to explore its action mechanism of alleviating H9C2 cell injury induced by high glucose.[Methods] Injured model of H9C2 cardiomyocyte was induced by high glucose. The normal control group,model control group and curcumin groups of different concentration (2.5,5,10,20 μmol/L) were set in this experiment. ELISA was used to determine the content of TNF-α,IL-6 and LDH in the cell culture supernatant,and contents of MDA and SOD in H9C2 cardiomyocyte. Western Blot was used to detect the protein expression levels of IKKβ,NF-κB p65 and p-NF-κB p65 in cells.[Results] High glucose treatment on H9C2 cardiomyocytes significantly down-regulated cell viability(P<0.05), the content of TNF-α,IL-6 and LDH in the cell culture supernatant were increased (P<0.05),the MDA content was increased and the SOD content was decreased in H9C2 cardiomyocyte (P<0.05);the protein expression of IKKβ,and p-NF-κB p65 were increased (P<0.05) in the model control group. Pretreatment with circumin (2.5,5,10,20 μmol/L) for 6h significantly maintained cell viability,reduced the content of TNF-α,IL-6 and LDH in the cell culture supernatant (P<0.05),and decreased the MDA content and increased the SOD content in H9C2 cardiomyocyte; down-regulated protein expression levels of IKKβ and p-NF-κB p65 (P<0.05).[Conclusion] Curcumin alleviates high glucose-induced H9C2 cardiomyocyte injury by inhibiting the excessive activation of IKK/NF-κB pathway,and alleviating inflammation and oxidative stress.