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

[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、20μmol/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-κB p65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。

[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(2.5,5,10,20μmol/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(2.5,5,10,20μmol/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 IKKβand 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.