摘要
目的:探讨狗脊多糖通过调节lncRNA NEAT1延缓椎间盘终板软骨细胞衰老的作用机制。方法:将15只C57BL/6小鼠的椎间盘终板软骨细胞进行体外培养;经10 ng·mL-1白细胞介素-1β(IL-1β)干预24 h诱导软骨细胞;将软骨细胞随机分为空白对照组、模型对照组(10 ng·mL-1IL-1β)和狗脊多糖各剂量组(100,200,400μg·mL-1);Real-time PCR及Western blot分别检测狗脊多糖对IL-1β诱导软骨细胞lncRNA NEAT1、衰老相关通路p53-p21中p53、p21水平及衰老相关分泌表型的基质金属蛋白酶-13(MMP-13)、CollagenⅡ、Caspase-3蛋白表达情况;流式细胞术检测各组软骨细胞凋亡率情况。结果:Real-time PCR结果显示,经IL-1β诱导的椎间盘终板软骨细胞中lncRNA NEAT1、p21、p53水平显著增高;与模型对照组比较,狗脊多糖各剂量组lncRNA NEAT1、p21、p53水平显著降低;与空白对照组比较,经IL-1β诱导的椎间盘终板软骨细胞中MMP-13、Caspase-3蛋白表达含量显著增高,CollagenⅡ蛋白表达降低;经狗脊多糖干预后,可显著改善MMP-13、CollagenⅡ、Caspase-3蛋白表达;此外,与模型对照组比较,狗脊多糖各剂量组椎间盘终板软骨细胞凋亡率呈现下降趋势。结论:狗脊多糖通过调控lncRNA NEAT1影响p53、p21、MMP-13、CollagenⅡ、Caspase-3表达,降低软骨细胞凋亡率,进而延缓椎间盘终板软骨细胞衰老。
Objective:To explore the mechanism of cibotium barometz polysaccharides in delaying the aging of intervertebral disc endplate chondrocytes by regulating lncRNA NEAT1.Methods:The intervertebral disc endplate chondrocytes of 15 C57BL/6 mice were cultured in vitro.10 ng·mL-1 of interleukin-1β(IL-1β)was used for intervention for 24 hours to induce chondrocytes.Chondrocytes were randomly divided into a blank control group and a model control group(10 ng·mL-1 IL-1β)and different dosage groups of cibotium barometz polysaccharides(100,200,400μg·mL-1).Real time PCR and Western blot were used to detect the effect of cibotium barometz polysaccharides on IL-1βinduced lncRNA NEAT1,the levels of p53,p21 in aging related pathway p53-p21 and the expression of aging related secretory phenotype MMP-13,CollagenⅡ,and Caspase-3 protein.Flow cytometry was used to detect the apoptosis rate of chondrocytes in each group.Results:Real time PCR results showed that the levels of lncRNA NEAT1,p21 and p53 in IL-1βinduced intervertebral disc endplate chondrocytes were significantly increased;compared with the model control group,the levels of lncRNA NEAT1,p21,and p53 in each dose group of cibotium barometz polysaccharides were significantly reduced;compared with the blank control group,the expression levels of MMP-13 and Caspase-3 proteins in IL-1βinduced intervertebral disc endplate chondrocytes were significantly increased,while the expression of CollagenⅡprotein was reduced.After intervention with cibotium barometz polysaccharides,the expression of MMP-13,CollagenⅡ,and Caspase-3 proteins could be significantly improved.In addition,compared with the model control group,the apoptosis rate of intervertebral disc endplate chondrocytes in the cibotium barometz polysaccharide group showed a decreasing trend.Conclusion:Cibotium barometz polysaccharides affect the expression of p53,p21,MMP-13,CollagenⅡ,and Caspase-3 by regulating lncRNA NEAT1,reducing the apoptosis rate of chondrocytes and delaying the aging of intervertebral disc
作者
付长龙
涂海水
张文键
陈毅涛
陆诗雨
李超
金灵璐
郑春松
FU Chang-long;TU Hai-shui;ZHANG Wen-jian;CHEN Yi-tao;LU Shi-yu;LI Chao;JIN Ling-lu;ZHENG Chun-song
出处
《风湿病与关节炎》
2024年第2期1-5,共5页
Rheumatism and Arthritis
基金
福建省自然科学基金项目(2022J01370)
福建中医药大学高层次人才科研启动资金项目(X2019011-人才)
陈可冀中西医结合发展基金资助项目(CKJ2022008)。
