Extracellular matrix(ECM)stiffening is a typical characteristic of cartilage aging,which is a quintessential feature of knee osteoarthritis(KOA).However,little is known about how ECM stiffening affects chondrocytes an...Extracellular matrix(ECM)stiffening is a typical characteristic of cartilage aging,which is a quintessential feature of knee osteoarthritis(KOA).However,little is known about how ECM stiffening affects chondrocytes and other molecules downstream.This study mimicked the physiological and pathological stiffness of human cartilage using polydimethylsiloxane(PDMS)substrates.It demonstrated that epigenetic Parkin regulation by histone deacetylase 3(HDAC3)represents a new mechanosensitive mechanism by which the stiffness matrix affected chondrocyte physiology.We found that ECM stiffening accelerated cultured chondrocyte senescence in vitro,while the stiffness ECM downregulated HDAC3,prompting Parkin acetylation to activate excessive mitophagy and accelerating chondrocyte senescence and osteoarthritis(OA)in mice.Contrarily,intra-articular injection with an HDAC3-expressing adeno-associated virus restored the young phenotype of the aged chondrocytes stimulated by ECM stiffening and alleviated OA in mice.The findings indicated that changes in the mechanical ECM properties initiated pathogenic mechanotransduction signals,promoted the Parkin acetylation and hyperactivated mitophagy,and damaged chondrocyte health.These results may provide new insights into chondrocyte regulation by the mechanical properties of ECM,suggesting that the modification of the physical ECM properties may be a potential OA treatment strategy.展开更多
Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control...Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing serum),and then 24h after intervention respectively,the cell proliferation and Apoptosis rate;Western blot detected the expression levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65,MMP-13 proteins in chondrocytes.ELISA detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1βgroup;compared with IL-1βgroup,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell proliferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-αlevel decreased.13 protein levels and TNF-αand IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclusion:BSBT-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,improve the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.展开更多
Objective This study aimed to investigate the potential mechanisms by which lysyl oxidase like 3(LOXL3)affects the autophagy in chondrocytes in osteoarthritis(OA),specifically through the activation of mammalian targe...Objective This study aimed to investigate the potential mechanisms by which lysyl oxidase like 3(LOXL3)affects the autophagy in chondrocytes in osteoarthritis(OA),specifically through the activation of mammalian target of rapamycin complex 1(mTORC1).Methods To establish an OA model,rats underwent anterior cruciate ligament transection(ACLT).Chondrocytes were isolated from cartilage tissues and cultured.Western blotting was performed to assess the expression of LOXL3,Rheb,phosphorylation of p70S6K(p-p70S6K,a downstream marker of mTORC1),and autophagy markers.The autophagy of chondrocytes was observed using an immunofluorescence assay.Results The expression levels of both LOXL3 and Rheb proteins were upregulated in chondrocytes isolated from the OA model cartilage,in comparison to those from the normal cartilage.The silencing of LOXL3 resulted in a decrease in the protein levels of Rheb and p-p70S6K,as well as an increase in the expression of autophagy-related proteins.Additionally,the effect of LOXL3 could be reversed through the silencing of Rheb.The results of the immunofluorescence assay confirmed the impact of LOXL3 and Rheb on chondrocyte autophagy.Conclusion LOXL3 inhibits chondrocyte autophagy by activating the Rheb and mTORC1 signaling pathways.展开更多
Endochondral ossification requires proper control of chondrocyte proliferation,differentiation,survival,and organization.Here we show that knockout ofα-parvin,an integrin-associated focal adhesion protein,from murine...Endochondral ossification requires proper control of chondrocyte proliferation,differentiation,survival,and organization.Here we show that knockout ofα-parvin,an integrin-associated focal adhesion protein,from murine limbs causes defects in endochondral ossification and dwarfism.The mutant long bones were shorter but wider,and the growth plates became disorganized,especially in the proliferative zone.With two-photon time-lapse imaging of bone explant culture,we provide direct evidence showing thatα-parvin regulates chondrocyte rotation,a process essential for chondrocytes to form columnar structure.Furthermore,loss ofα-parvin increased binucleation,elevated cell death,and caused dilation of the resting zones of mature growth plates.Single-cell RNA-seq analyses revealed alterations of transcriptome in all three zones(i.e.,resting,proliferative,and hypertrophic zones)of the growth plates.Our results demonstrate a crucial role ofα-parvin in long bone development and shed light on the cellular mechanism through whichα-parvin regulates the longitudinal growth of long bones.展开更多
Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartil...Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartilage remains themajor hallmark of the disease. Current treatments targeting OA symptoms have a limited impact on impeding orreversing the OA progression. Understanding the molecular and cellular mechanisms underlying OA development isa critical barrier to progress in OA therapy. Recent studies by the current authors’ group and others have revealedthat the nuclear factor of activated T cell 1 (NFAT1), a member of the NFAT family of transcription factors, regulatesthe expression of many anabolic and catabolic genes in articular chondrocytes of adult mice. Mice lacking NFAT1exhibit normal skeletal development but display OA in both appendicular and spinal facet joints as adults. Thisreview mainly focuses on the recent advances in the regulatory role of NFAT1 transcription factor in the activities ofarticular chondrocytes and its implication in the pathogenesis of OA.展开更多
背景:有研究发现,软骨细胞铁死亡可作为重要的发病机制诱导骨关节炎的发生与发展。随着祖国医学的不断发展,有学者发现某些中药单体、中药复方等可通过多种作用机制抑制软骨细胞发生铁死亡,最终起到治疗骨关节炎的作用。目的:探讨铁死...背景:有研究发现,软骨细胞铁死亡可作为重要的发病机制诱导骨关节炎的发生与发展。随着祖国医学的不断发展,有学者发现某些中药单体、中药复方等可通过多种作用机制抑制软骨细胞发生铁死亡,最终起到治疗骨关节炎的作用。目的:探讨铁死亡与骨关节炎的关系及中医药干预铁死亡治疗骨关节炎的作用机制,以期为骨关节炎的治疗及预防提供新的思路。方法:以“铁死亡,骨关节炎,软骨细胞,脂质代谢,中医药,活性氧,谷胱甘肽过氧化物酶”为中文检索词,以“osteoarthritis,ferroptosis,chondrocytes,lipid metabolism,traditional Chinese medicine,ROS,GPX4”为英文检索词,检索中国知网、PubMed、万方、维普等数据库,筛选各数据库建库至2022年铁死亡与骨关节炎及其中医药干预研究的相关文章,并进行系统的整理分析。结果与结论:①细胞内铁超载会引发脂质过氧化反应,造成活性氧积累,最终导致铁死亡的发生,而铁过量会加速软骨细胞凋亡,影响软骨细胞的稳态,进而诱导骨关节炎的发生。②氨基酸代谢异常时,谷氨酸与胱氨酸交换失衡,导致谷氨酸在细胞外积聚,进而导致谷胱甘肽被耗尽,间接抑制了铁死亡相关因子的功能,对铁死亡起到促进作用。③细胞脂质代谢异常时,细胞膜的重要组成部分不饱和脂肪酸失去双烯丙基氢原子,导致细胞被破坏诱发铁死亡,而脂质代谢与骨关节炎的发生发展密切相关。④中医药可以通过干预诱导铁死亡发生的相关机制对骨关节炎起到防治作用。目前对铁死亡的研究仍不成熟,继续深入研究其作用机制,有望为临床治疗骨关节炎开辟新途径。展开更多
基金supported by the National Key R&D Program of China(2022YFB4600600)National Natural Science Foundation of China(32271181,82301785)+1 种基金Guangdong Basic and Applied Basic Research Foundation(2020B1515120001)Sichuan Province Central Guided Local Science and Technology Development Special Program(2023ZYD0072).
文摘Extracellular matrix(ECM)stiffening is a typical characteristic of cartilage aging,which is a quintessential feature of knee osteoarthritis(KOA).However,little is known about how ECM stiffening affects chondrocytes and other molecules downstream.This study mimicked the physiological and pathological stiffness of human cartilage using polydimethylsiloxane(PDMS)substrates.It demonstrated that epigenetic Parkin regulation by histone deacetylase 3(HDAC3)represents a new mechanosensitive mechanism by which the stiffness matrix affected chondrocyte physiology.We found that ECM stiffening accelerated cultured chondrocyte senescence in vitro,while the stiffness ECM downregulated HDAC3,prompting Parkin acetylation to activate excessive mitophagy and accelerating chondrocyte senescence and osteoarthritis(OA)in mice.Contrarily,intra-articular injection with an HDAC3-expressing adeno-associated virus restored the young phenotype of the aged chondrocytes stimulated by ECM stiffening and alleviated OA in mice.The findings indicated that changes in the mechanical ECM properties initiated pathogenic mechanotransduction signals,promoted the Parkin acetylation and hyperactivated mitophagy,and damaged chondrocyte health.These results may provide new insights into chondrocyte regulation by the mechanical properties of ECM,suggesting that the modification of the physical ECM properties may be a potential OA treatment strategy.
基金National Natural Science Foundation of China(No.82360934)Science and Technology Innovation Leading Talents Project of Xinjiang Uygur Autonomous Region(No.2022TSYCLJ0007)+1 种基金Xinjiang Uygur Autonomous Region Key Research and Development Task Special Project(No.2021B03006)Natural Science Foundat ion of Xinj iang Uygur Autonomous Region(No.2022D01C170,2022D01C171)。
文摘Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing serum),and then 24h after intervention respectively,the cell proliferation and Apoptosis rate;Western blot detected the expression levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65,MMP-13 proteins in chondrocytes.ELISA detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1βgroup;compared with IL-1βgroup,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell proliferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-αlevel decreased.13 protein levels and TNF-αand IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclusion:BSBT-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,improve the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.
基金the National Natural Science Foundation of China(No.81702187)Natural Science Foundation of Jiangxi Province(No.20202BAB206019)+4 种基金Science Fund for Distinguished Young Scholars of Jiangxi Province(No.20224ACB216018)Scientific Talents Grants of Jiangxi Province(No.S2018LQCQ0800)Scientific Grants of Health Commission of Jiangxi Province(No.20194048)Scientific Innovation Talents Grants of Ganzhou(No.2019-60-08)Leading Talents Grants and Ph.D.Programs Foundation of Ganzhou People’s Hospital(No.Bsqd2019003)and Academic leaders Program of Ganzhou Institutes of Health.
文摘Objective This study aimed to investigate the potential mechanisms by which lysyl oxidase like 3(LOXL3)affects the autophagy in chondrocytes in osteoarthritis(OA),specifically through the activation of mammalian target of rapamycin complex 1(mTORC1).Methods To establish an OA model,rats underwent anterior cruciate ligament transection(ACLT).Chondrocytes were isolated from cartilage tissues and cultured.Western blotting was performed to assess the expression of LOXL3,Rheb,phosphorylation of p70S6K(p-p70S6K,a downstream marker of mTORC1),and autophagy markers.The autophagy of chondrocytes was observed using an immunofluorescence assay.Results The expression levels of both LOXL3 and Rheb proteins were upregulated in chondrocytes isolated from the OA model cartilage,in comparison to those from the normal cartilage.The silencing of LOXL3 resulted in a decrease in the protein levels of Rheb and p-p70S6K,as well as an increase in the expression of autophagy-related proteins.Additionally,the effect of LOXL3 could be reversed through the silencing of Rheb.The results of the immunofluorescence assay confirmed the impact of LOXL3 and Rheb on chondrocyte autophagy.Conclusion LOXL3 inhibits chondrocyte autophagy by activating the Rheb and mTORC1 signaling pathways.
基金supported by the National Natural Science Foundation of China Grant 82273308,Inno HK@Health,Theme-based Research Scheme (Tl3-602/21-N)Guangdong-Dongguan Joint Research Scheme Guangdong-Hong Kong-Macao Program (2021B1515130004)+4 种基金the Natural Science Foundation of Guangdong Province Grant 2017B030301018the Special Support Program for Training High-Level Talents in Guangdong Grant 2019TQ05Y518the Shenzhen Innovation Committee of Science and Technology Grant JCYJ20220530112817040,ZDSYS20220606101604009the National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital&Shenzhen Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Shenzhen E010122002supported by the Lombardi and Shinozuka Experimental Pathology Research Endowment Fund,University of Pittsburgh。
文摘Endochondral ossification requires proper control of chondrocyte proliferation,differentiation,survival,and organization.Here we show that knockout ofα-parvin,an integrin-associated focal adhesion protein,from murine limbs causes defects in endochondral ossification and dwarfism.The mutant long bones were shorter but wider,and the growth plates became disorganized,especially in the proliferative zone.With two-photon time-lapse imaging of bone explant culture,we provide direct evidence showing thatα-parvin regulates chondrocyte rotation,a process essential for chondrocytes to form columnar structure.Furthermore,loss ofα-parvin increased binucleation,elevated cell death,and caused dilation of the resting zones of mature growth plates.Single-cell RNA-seq analyses revealed alterations of transcriptome in all three zones(i.e.,resting,proliferative,and hypertrophic zones)of the growth plates.Our results demonstrate a crucial role ofα-parvin in long bone development and shed light on the cellular mechanism through whichα-parvin regulates the longitudinal growth of long bones.
基金supported by the United States National Institutes of Health(NIH)under Award Number R01 AR059088(to J.W.)the Mary A.and Paul R.Harrington Distinguished Professorship Endowment.
文摘Osteoarthritis (OA), the most common form of joint disease, is characterized clinically by joint pain, stiffness,and deformity. OA is now considered a whole joint disease;however, the breakdown of the articular cartilage remains themajor hallmark of the disease. Current treatments targeting OA symptoms have a limited impact on impeding orreversing the OA progression. Understanding the molecular and cellular mechanisms underlying OA development isa critical barrier to progress in OA therapy. Recent studies by the current authors’ group and others have revealedthat the nuclear factor of activated T cell 1 (NFAT1), a member of the NFAT family of transcription factors, regulatesthe expression of many anabolic and catabolic genes in articular chondrocytes of adult mice. Mice lacking NFAT1exhibit normal skeletal development but display OA in both appendicular and spinal facet joints as adults. Thisreview mainly focuses on the recent advances in the regulatory role of NFAT1 transcription factor in the activities ofarticular chondrocytes and its implication in the pathogenesis of OA.
文摘背景:有研究发现,软骨细胞铁死亡可作为重要的发病机制诱导骨关节炎的发生与发展。随着祖国医学的不断发展,有学者发现某些中药单体、中药复方等可通过多种作用机制抑制软骨细胞发生铁死亡,最终起到治疗骨关节炎的作用。目的:探讨铁死亡与骨关节炎的关系及中医药干预铁死亡治疗骨关节炎的作用机制,以期为骨关节炎的治疗及预防提供新的思路。方法:以“铁死亡,骨关节炎,软骨细胞,脂质代谢,中医药,活性氧,谷胱甘肽过氧化物酶”为中文检索词,以“osteoarthritis,ferroptosis,chondrocytes,lipid metabolism,traditional Chinese medicine,ROS,GPX4”为英文检索词,检索中国知网、PubMed、万方、维普等数据库,筛选各数据库建库至2022年铁死亡与骨关节炎及其中医药干预研究的相关文章,并进行系统的整理分析。结果与结论:①细胞内铁超载会引发脂质过氧化反应,造成活性氧积累,最终导致铁死亡的发生,而铁过量会加速软骨细胞凋亡,影响软骨细胞的稳态,进而诱导骨关节炎的发生。②氨基酸代谢异常时,谷氨酸与胱氨酸交换失衡,导致谷氨酸在细胞外积聚,进而导致谷胱甘肽被耗尽,间接抑制了铁死亡相关因子的功能,对铁死亡起到促进作用。③细胞脂质代谢异常时,细胞膜的重要组成部分不饱和脂肪酸失去双烯丙基氢原子,导致细胞被破坏诱发铁死亡,而脂质代谢与骨关节炎的发生发展密切相关。④中医药可以通过干预诱导铁死亡发生的相关机制对骨关节炎起到防治作用。目前对铁死亡的研究仍不成熟,继续深入研究其作用机制,有望为临床治疗骨关节炎开辟新途径。
