【摘要】 目的探讨二氢杨梅素(DHM)对高糖(HG)诱导的心肌细胞H9C2损伤的影响及机制。方法细胞处理分为对照组、35 mmol/L HG组、35mmol/L HG+50μmol/L DHM组及50μmol/L DHM组。CCK-8法检测细胞活力,化学比色法检测丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)水平,流式细胞术检测ROS水平;荧光定量PCR法及Elisa法分别检测TNF-α、IL-1β、IL-6 mRNA和含量,Western Blotting检测p-IκBα、IκBα蛋白及核蛋白NF-κB p65的表达水平。采用单因素方差分析进行组间比较。结果对照组、35mmol/L HG组、35 mmol/L HG+50μmol/L DHM组、35 mmol/L HG+100μmol/L DHM组的细胞活力分别是(100±0.00)%、(52.23±5.69)%、(74.58±6.12)%和(86.04±3.76)%,差异具有统计学意义(F=40.61,P <0.01)。对照组、35 mmol/L HG组和35 mmol/L HG+100μmol/L DHM组的MDA和ROS水平,SOD和CAT活性分别是(0.44±0.06) nmol/ml,(2.33±0.40) nmol/ml,(1.48±0.41) nmol/ml、(156.0±9.00)U/ml,(325.3±10.69)U/ml,(244.0±9.54) U/ml,(10.62±1.59) U/ml,(5.18±0.34)U/ml,(7.75±0.53)U/ml,(11.31±0.98) U/ml,(5.20±1.12) U/ml和(8.06±0.66)U/ml,差异具有统计学意义(F=30.34,29.75,14.72,P均<0.01)。DHM预处理可明显拮抗HG对H9C2心肌细胞TNF-α、IL-1β和IL-6 mRNA及含量的上调作用,差异存在统计学意义(P均<0.01)。DHM可抑制HG对H9C2心肌细胞p-IκBα/IκBα蛋白和核蛋白NF-κB p65表达的增加作用,差异存在统计学意义(P均<0.01)。结论 DHM可拮抗HG诱导的H9C2心肌细胞损伤,这可能与其抑制NF-κB信号通路有关。更多还原

【Abstract】 Objective To investigate the influence of dihydrobayberry(DHM)on high glucose(HG)-induced cardiomyocytes H9 χ~2 cells damage and its poteintial mechanism.Methods Cells were divided into the following groups: control group, 35 mmol/L HG group,35 mmol/L HG+50 μmol/L DHM group and 50 μmol/L DHM group. CCk-8 assay was used to detect cell viability. The levels of malondialdehyde(MDA), superoxide dismutase(SOD) and catalase(CAT) were determined by chemical colorimetry. ROS levels were measured by flow cytometry.TNF-α, IL-1β and IL-6 mRNA expression and contents were determined by fluorescence quantitative PCR and Elisa assays respectively. The expression levels of p-IκBα and IκBα proteins, and nucleoprotein NF-κB p65 were detected by Western Blot. Univariate analysis of variance was used for comparison between groups. Results Pretreatment with 50 and 100 μmol/L DHM significantly inhibited the reduced cell viability of H9 C2 myocardial cells caused by 35 mmol/L HG: The cell viability of the control group, 35 mmol/L HG group, 35 mmol/L HG+50 μmol/L DHM group,35 mmol/L HG+100 μmol/L DHM group were(100±0.00)%,(52.23±5.69)%,(74.58±6.12)%and(86.04±3.76)%, respectively(F = 40.61, P < 0.001). We also found that pretreatment with DHM(50 μmol/L)significantly inhibited the enhanced MDA and ROS levels, and decreased SOD and CAT activity of H9 C2 myocardial cells induced by HG(35 mmol/L). The MDA level, SOD and CAT activity of the control group, HG group and HG+ DHM group were(0.44±0.06) nmol/ml,(2.33±0.40)nmol/ml,(1.48±0.41)nmol/ml,(156.00±9.00)U/ml,(325.3±10.69)U/ml,(244.0±9.54)U/ml,(10.62±1.59)U/ml,(5.18±0.34)U/ml,(7.75±0.53) U/ml and(11.31±0.98) U/ml,(5.20±1.12)U/ml,(8.06±0.66)U/ml, respectively.(F = 30.34, 29.75,14.72, P all < 0.001). Pretreatment with DHM significantly inhibited the increased expression levels of TNF-α, IL-1β and IL-6 mRNA and their contents caused by HG in H9 C2 myocardial cells(P all <0.001). DHM significantly inhibited the increased expression of p-IκBα/IκBα and nucleoprotein NF-κB p65 in H9 C2 myocardial cells caused by HG(P all < 0.001). Conclusion DHM can antagonize HG-induced H9 C2 myocardial cell injury, which may be related to its inhibition of NF-κB signaling pathway.更多还原