兰州大学学报（医学版）2023，Vol.49Issue(11)：9-17,25,10.DOI:10.13885/j.issn.1000-2812.2023.11.002
原花青素对过氧化氢诱导羊颞下颌关节盘细胞氧化损伤的保护作用研究
The shielding power of proanthocyanidins against oxidative harm caused by hydrogen peroxide in sheep temporomandibular joint disc cells
摘要
Abstract
Objective To investigate the protective power of proanthocyanidins against oxidative harm caused by hydrogen peroxide in temporomandibular joint disc cells.Methods Temporomandibular joint disc cells were treated with concentrations of 0,100,200,400 and 800 μmol/L hydrogen peroxide for 12,24 and 48 h,respectively,and cell viability detected by CCK-8.Then temporomandibular joint disc cells were treated with different concentrations of proanthocyanidins(0,5,10,20,40 μmol/L)for 24,48 h,and CCK-8 assayed the effect of proanthocyanidins on temporomandibular joint disc cell viability.Then the experiments were divided into the control group,model group,low concentration group(concentration of 5 μmol/L proanthocyanidins),medium concentration group(concentration of 10 μmol/L proanthocyanidins)and high concentration group(concentration of 20 μmol/L proanthocyanidins).Cell viability was detected using the CCK-8 method,while levels of malondialdehyde and superoxide dismutase were detected using colorimetric assay.Reactive oxygen species were detected using DCFH-DA fluorescent probe staining,apoptosis was measured using flow cytometry,and mRNA expression levels of inflammatory factors,extracellular matrix anabolic factors,and extracellular matrix catabolic factors were detected using quantitative reverse transcription PCR.Results Cell viability was 60.57%when temporomandibular joint disc cells were treated with 400 μmol/L hydrogen perox-ide for 24 h,and compared with the control group,the cell survival rate decreased(P<0.05).Compared with the control group,the model group showed decreased temporomandibular joint disc cells viability,increased levels of reactive oxygen species and malondialdehyde,and decreased activity of superoxide dismutase.Compared to the model group,the 5,10 and 20 μ mol/L proanthocyanidins treatment groups can increase temporomandibular joint disc cells viability induced by hydrogen peroxide,reduce the levels of reactive oxygen species and malondialdehyde,and increase the activity of intracellular superoxide dismutase(P<0.05).The flow results revealed a higher level of apoptosis in the model group than in the control group,yet this was diminished post-proanthocyanidins pretreatment(P<0.05).Moreover,the expression of intracellular inflam-matory factors(interleukin-1β,tumornecrosis factor-α,interleukin-6)was higher in the model group,whereas the expression of interleukin-1β,tumornecrosis factor-α,interleukin-6 was reduced post-proanthocyanidins pretreatment(P<0.05).The expression of typeⅠcollagen and aggrecan was augmented by proanthocyanidins when hydrogen peroxide was induced(P<0.05),while the expression of matrix metalloproteinase-1,matrix metalloproteinase-3 and matrix metalloproteinase-9 was inhibited(P<0.05)and extracellular matrix degrada-tion suppressed.Conclusion The most effective action condition for constructing an oxidative stress model of temporomandibular joint disc cells was 400 μmol/L hydrogen peroxide for 24 h.Proanthocyanidins reversed hydrogen peroxide-induced lipid peroxidation,reactive oxygen species generation,inflammatory damage and extracellular matrix degradation in temporomandibular joint disc cells,and inhibited oxidative stress injury by increasing antioxidant enzyme levels and attenuating apoptosis.关键词
颞下颌关节盘细胞/原花青素/过氧化氢/氧化应激/凋亡/细胞外基质Key words
temporomandibular joint disc cells/proanthocyanidins/hydrogen peroxide/oxidative stress/apop-tosis/extracellular matrix分类
医药卫生引用本文复制引用
胡静静,包广洁,康宏..原花青素对过氧化氢诱导羊颞下颌关节盘细胞氧化损伤的保护作用研究[J].兰州大学学报（医学版）,2023,49(11):9-17,25,10.基金项目
甘肃省自然科学基金资助项目(21JR7RA161) （21JR7RA161）
兰州大学口腔医学院基金资助项目(533000/071100122) （533000/071100122）
