摘要
目的通过建立H2O2诱导软骨细胞损伤模型,观察姜黄素对软骨细胞的保护作用。方法体外培养SD大鼠关节软骨细胞,随机分为对照组、模型组(H2O2)、姜黄素低中高剂量剂量组(20、40、80μmol/L)。姜黄素与软骨细胞培养48 h后,加入H2O2,24 h后收集细胞,MTT和流式细胞仪测定细胞增殖能力,测定细胞超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)含量,Real-Time PCR和Western blot法检测细胞Nrf2 mRNA及其蛋白表达水平。结果与对照组比较,模型组软骨细胞存活率降低,具有分裂象的S期和G2/M期细胞明显减少;SOD、CAT含量降低,MDA含量增加,Nrf2 mRNA及其蛋白表达水平下调。经姜黄素预先处理,软骨细胞存活率升高,细胞增殖能力恢复,SOD、CAT含量升高,MDA含量降低,Nrf2 mRNA及其蛋白表达水平升高。随浓度升高变化更明显。结论姜黄素对软骨细胞氧化应激损伤有保护作用,可通过增强Nrf2表达实现。
Objective To observe the protective effects of curcumin on chondrocytes through setting up the models of chondrocytes injury of rats in vitro induced by H 2 O2 K.Methods The articular chondrocytes of SD rats were prepared and cultured in vitro,which were randomly divided into five groups：control group, model group （H2O2）, low-dose curcumin group （20μmol/L）,median-dose curcumin group （40μmol/L） and high-dose curcumin group （80μmol/L）.On 48 hours after the articular chondrocytes were cultured in culture medium with curcumin , H2 O2 was added into the culture medium and co-cultured for 24 hous, then the articular chondrocytes were collected .The cell proliferation ability and the contents of superoxide dismutase （SOD）,malondialdehyde （MDA）,catalase （CAT） were detected by MTT assay and flow cytometry , respectively.The expression levels of Nrf2 mRNA and protein were detected by Real-time PCR and Western Blot,respectively. Results As compared with those in control group ,the survival rates of chondrocytes in model group were decreased ,and the numbers of chondrocytes with mitotic figure in stage S and G 2/M were obviously decreased .The contents of SOD and CAT were decreased,however,the levels of MDA were increased.The expression levels of Nrf2 mRNA and protein were down-regulated. After the chondrocytes were pretreated by curcumin , the survival rates of chondrocytes were increased , the cell proliferation ability was improved , and the contents of SOD and CAT were increased , however , the levels of MDA were decreased .The expression levels of Nrf2 mRNA and protein were increased with the increase of concentration of curcumin .Conclusion The curcumin has protective effects on oxidative stress injury of chondrocytes by increasing the expression levels of Nrf 2.
出处
《河北医药》
CAS
2016年第23期3525-3528,共4页
Hebei Medical Journal
基金
河北省医学科学研究重点课题(编号:20150666)
中国博士后科学基金资助项目(编号:2015M582480)
广东省自然科学基金项目(编号:2016A030313100)
