姜黄素对H2O2致H9c2心肌细胞损伤的抗凋亡作用及对NF-κB信号通路的调节作用研究
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篇名: 姜黄素对H2O2致H9c2心肌细胞损伤的抗凋亡作用及对NF-κB信号通路的调节作用研究
TITLE: Study on the Anti-apoptotic Effect of Curcumin on H 2O2-induced H 9c2 Cardiomyocyte Injury and the Regulation of NF-κB Signaling Pathway
摘要: 目的:探讨姜黄素对H2O2致H9c2心肌细胞损伤的抗凋亡作用及对核因子κB(NF-κB)信号通路的调节作用。方法:将大鼠H9c2心肌细胞随机分为正常对照组、损伤模型组和姜黄素低、中、高剂量组(25、50、100μmol/L)。正常对照组不作任何干预;损伤模型组细胞以50μmol/LH2O2诱导12h以建立损伤模型;姜黄素各剂量组经相应浓度药物预处理24h后,再以50μmol/LH2O2诱导12h。继续培养24h后,采用MTT法和TUNEL法分别检测细胞的存活率和凋亡率;采用WTS-8法和显色法分别检测细胞中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;采用免疫荧光法检测细胞中微管相关蛋白1轻链3(LC3)阳性表达的相对荧光强度;采用实时定量聚合酶链式反应法检测细胞中NF-κBp65mRNA的表达水平;采用Westernblotting法检测细胞中NF-κBp65、磷酸化NF-κBp65(p-NF-κBp65)蛋白的表达水平。结果:与正常对照组比较,损伤模型组细胞的存活率和SOD活性均显著降低,细胞凋亡率、MDA含量、LC3阳性表达相对荧光强度以及NF-κBp65mRNA和NF-κBp65、p-NF-κBp65蛋白的表达水平以及p-NF-κB/NF-κB比值均显著升高(P<0.05)。与损伤模型组比较,姜黄素各剂量组细胞存活率和SOD活性均显著升高,细胞凋亡率、MDA含量、LC3阳性表达相对荧光强度以及NF-κBp65mRNA和NF-κB/p65、p-NF-κBp65蛋白的表达水平以及p-NF-κBp65/NF-κBp65比值均显著降低(P<0.05)。结论:姜黄素能够提高H2O2致损伤H9c2心肌细胞的存活率、降低其凋亡率,提高心肌细胞中SOD活性并降低MDA含量;上述作用可能与调节心肌细胞NF-κB信号通路有关。
ABSTRACT: OBJECTIVE:To investigate the anti-apoptotic effect of curcumin on H 2O2-induced H 9c2 cardiomyocyte injury and the regulation of NF-κB signaling pathway. METHODS:H9c2 cardiomyocyte were randomly divided into normal control group , injury model group ,curcumin low-dose ,medium-dose and high-dose groups (25,50,100 μmol/L). Normal control group didn ’t received any intervention. The cells in injury model group were induced with 50 μmol/L H2O2 for 12 h to establish the injury model. The cells in curcumin groups were treated with relevant concentration of drugs for 24 h,and then induced with 50 μmol/L H2O2 for 12 h. After cultured for 24 h,survival rate and apoptotic rate of cells were measured by MTT method and TUNEL method ;SOD activity and MDA content were determined by WTS- 8 assay and color test ;relative fluorescence intensity of LC 3 positive expression was detected by immunofluorescence method ;mRNA expression of NF-κB p65 in cells was detected by real-time PCR ; Western blotting assay was used to detect the protein expression of NF-κB p65 and p-NF-κB p65 in cells. RESULTS :Compared with normal control group ,survival rate and SOD activity were decreased significantly in injury model group ,while apoptotic rate , MDA content ,relative fluorescence intensity of LC 3 positive expression ,mRNA expression of NF-κB,protein expression of NF-κ B p 65 and p-NF-κB p65 as well as p-NF-κB/NF-κB were increased significantly(P<0.05). Compared with injury model group , survival rates and SOD activities were increased significantly in curcumin groups ,while apoptotic rates ,MDA contents ,relative fluorescence intensities of LC 3 positive expression ,mRNA expression of LC 3 positive cells ,protein expression of NF-κB p65 and p-NF-κ B p65 as well as p-NF-κ B p65/NF-κ B p65 were decreased significantly (P<0.05). CONCLUSIONS :Curcumin can increase the survival rate of H 2O2-induced H 9c2 cardiomyocyte injury ,decrease its apoptotic rate ,increase SOD activity and decrease MDA content in cardiomyocytes. Above effects may be related to the regulation of NF-κB signaling pathway.
期刊: 2020年第31卷第23期
作者: 顾长海,李忠,陈冬玲
AUTHORS: GU Changhai ,LI Zhong ,CHEN Dongling
关键字: 姜黄素;H9c2细胞;心肌细胞损伤;抗凋亡作用;核因子κB信号通路
KEYWORDS: Curcumin;H9c2 cells;Cardiomyocyte injury ;Anti-apoptotic effect ;NF-κB signaling pathway
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