畜牧兽医学报2024，Vol.55Issue(8)：3714-3724,11.DOI:10.11843/j.issn.0366-6964.2024.08.043
白果内酯调控AMPK-SIRT3正反馈环路介导的线粒体生物发生改善ATDC5软骨细胞炎性损伤
Bilobalide Regulates Mitochondrial Biogenesis Mediated by AMPK-SIRT3 Positive Feedback Loop and Improves Inflammatory Damage of ATDC5 Chondrocytes
摘要
Abstract
This study takes the AMPK-SIRT3 positive feedback loop as the starting point.The purpose is to explore the mechanism of improvement of bilobalide on inflammatory damage in-duced interleukin-1β(IL-1β)in ATDC5 chondrocytes.Using IL-1β to induce inflammatory dam-age of ATDC5 chondrocytes to construct an osteoarthritis model in vitro.Randomly divided into 5 groups(a control group,IL-1β group,IL-1β group cotreated with bilobalide)which cotreat-ment group is further divided into low,medium,and high dose groups based on the application concentration of bilobalide(15,30 and 60 μmol·L-1).Western blot and real-time fluorescence quantitative PCR(qRT-PCR)methods were used to detect the protein and mRNA expression of ADAMTS4,PGC-1a,Collagen Type Ⅱ,MMP-3,NRF-1,and Fis1 in each group.A reagent kit was used to detect the ATP content in each group.The expression level of SIRT3 was detected by immunofluorescence.Western blot was used to detect the levels of AMPK-SIRT3 positive feedback loop related proteins(p-AMPK,AMPK,and SIRT3).Construct an AMPK-SIRT3 sig-naling pathway blockade model by treating ATDC5 chondrocytes with Compound C and 3-TYP.Mainly detecting downstream related protein changes(PGC-1a,NRF-1,and Fis1).The results showed that bilobalide down-regulated the expression of ADAMTS4 and MMP-3(P＜0.05),promoted the expression of Type Ⅱ collagen(P＜0.05),regulated ECM metabolic balance,and promoted ATP synthesis.Mechanistically,after intervention with bilobalide,p-AMPK,SITR3,PGC-1a and NRF-1 levels(P＜0.05)significantly increased while Fis1 levels(P＜0.05)signifi-cantly decreased.After pre-treatment with Compound C and 3-TYP in ATDC5 chondrocytes,PGC-1a,NRF-1 and Fis1 protein levels were inhibited to varying degrees.In summary,bilob-alide activates the expression of PGC-1a through the AMPK-SIRT3 positive feedback loop and regulates mitochondrial biogenesis to improve inflammatory damage in ATDC5 chondrocytes.关键词
软骨细胞/炎性损伤/白果内酯/AMPK-SIRT3正反馈环路/线粒体生物发生Key words
chondrocytes/inflammatory damage/bilobalide/AMPK-SIRT3 positive feedback loop/mitochondrial biogenesis分类
农业科技引用本文复制引用
李亚楠,马天文,马玉辉,魏成威..白果内酯调控AMPK-SIRT3正反馈环路介导的线粒体生物发生改善ATDC5软骨细胞炎性损伤[J].畜牧兽医学报,2024,55(8):3714-3724,11.基金项目
中国博士后科学基金项目(2023TQ0053) （2023TQ0053）
黑龙江省自然科学基金项目(LH2023C025 （）
YQ2023C015) （）
