scholarly journals Baicalin Promotes Chondrocyte Viability and the Synthesis of Extracellular Matrix through TGF-β/Smad3 Pathway in Mouse Chondrocytes

2021 ◽  
Author(s):  
Pengzhen Wang ◽  
Shaoheng Zhang ◽  
Chaosheng Yu ◽  
Xifeng Xiong ◽  
Aiguo Li ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Nuclear Translocation ◽  
Type Ii Collagen ◽  
The Elderly ◽  
Joint Disease ◽  
Down Regulation ◽  
Chondrocyte Viability ◽  
Scutellaria Baicalensis Georgi ◽  
Smad3 Phosphorylation ◽  
Flavonoid Monomer

Abstract Background: Osteoarthritis (OA) is epidemic in the elderly people as a common chronic joint disease. By now, drug and surgical treatments are two main therapies for OA worldwidely. Baicalin (BA) is a flavonoid monomer extracted from Scutellaria baicalensis Georgi and is reported that BA has anti-inflammatory, anti-deformation and anti-bacterial effects. Methods: Micromass culture, alcian blue and Safran O (SO)/fast green staining were used to investigate chondrocyte viability and ECM synthesis in chondrocytes of all groups. The expression of SOX9 and Smad3 in chondrocytes of all groups were detected by western blot and RT-qPCR, the expression of aggrecan (AGG), type II collagen (Col2α), MMP9/13 and ADAMTS5 were detected by RT-qPCR. In current study, we demonstrated that BA neutralized the down-regulation of chondrocyte viability and extracellular matrix (ECM) secretion, including AGG and Col2α, induced by IL-1β. As the key regulators of ECM, the down-regulation of SOX9, and the up-regulation of MMP9/13 and ADAMTS5 induced by IL-1β were abolished by BA. Moreover, BA increased the nuclear translocation and phosphorylation of Smad3, one key mediator of TGF-β/Smads pathway. Interestingly, the addition of Smad3 inhibitor SIS3 reversed the promotions of BA on chondrocyte viability, ECM secretion, SOX9 expression, Smad3 nuclear translocation and Smad3 phosphorylation, and the down-regulation of BA on the expressions of MMP9/13 and ADAMTS5. Conclusions: These results imply that BA can protect chondrocytes against IL-1β-induced inflammatory injury through the acceleration of Smad3 phosphorylation and nuclear translocation in chondrocytes. This study demonstrates that BA may be a potential drug for OA clinical treatment.

scholarly journals Kelainan Matriks Ekstraseluler Agrekan pada Osteoarthritis

2020 ◽  
Vol 9 (1) ◽  
pp. 67-80
Author(s):  
Umiatin Umiatin ◽  
Jeanne A Diwinata Pawitan
Keyword(s):  
Extracellular Matrix ◽  
Enzyme Activity ◽  
Molecular Mechanisms ◽  
Cartilage Damage ◽  
Type Ii Collagen ◽  
Joint Disease ◽  
Joint Cartilage ◽  
Keywords Osteoarthritis ◽  
A Disintegrin And Metalloproteinase ◽  
Thrombospondin Motif

Abstrak Osteoarthritis (OA) merupakan penyakit sendi dengan prevalensi paling tinggi yang menyebabkan nyeri kronis dan disabilitas. Berbagai faktor antara lain faktor mekanik, biokimia dan faktor enzimatik berperan dalam perkembangan OA. Perkembangan OA dicirikan oleh degradasi berlebihan pada agrekan dalam matriks ekstraseluler tulang rawan sendi. Agrekan berfungsi menyediakan fleksibilitas, viskoelastisitas dan kompresibilitas jaringan. Struktur agrekan tidak konstan sepanjang hidup, namun mengalami perubahan yang disebabkan oleh aktivitas sintesis maupun degradasi. Degradasi agrekan merupakan penanda awal kerusakan tulang rawan sendi pada OA, yang diikuti oleh kerusakan kolagen tipe II. Sejauh ini mekanisme molekulernya belum diketahui pasti, sehingga diperlukan penelitian lebih lanjut mengenai mekanisme dan penyebab kerusakan agrekan. Tulisan ini merupakan suatu kajian naratif berdasarkan artikel dari jurnal nasional dan internasional yang bertujuan untuk memberikan informasi mengenai agrekan meliputi struktur, fungsi, dan faktor-faktor yang berperan pada perubahan struktur agrekan yang menginduksi terjadinya OA. Hasil kajian menunjukkan bahwa perubahan struktur agrekan erat kaitannya dengan perubahan fungsi mekanik tulang rawan sendi. Perubahan ini terjadi terutama karena degradasi yang disebabkan oleh aktivitas enzim, dari keluarga matriks metalloprotease (MMP) dan a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS). Dari kajian ini disimpulkan bahwa degradasi agrekan karena aktivitas enzim berperan penting dalam perkembangan OA, sehingga perlu dilakukan penelitian untuk mencari inhibitor enzim MMP dan ADAMTS sebagai agen terapeutik untuk menghambat perkembangan dan progresivitas OA. Kata kunci: osteoarthritis, matriks ektraseluler, agrekan, degradasi. Abstract Osteoarthritis (OA) is a joint disease with the highest prevalence and a major cause of chronic pain and disability. Many factors such as mechanical, biochemical, and enzymatic factors are involved in OA development. The development of OA is characterized by excessive degradation of aggrecan in the extracellular matrix of articular cartilage, which functions to provide flexibility, viscoelasticity, and tissue compressibility. The structure of aggrecan is not constant throughout life but undergoes changes caused by synthesis and degradation activities. Aggrecan degradation is an early marker of joint cartilage damage in OA, followed by type II collagen damage. However, the molecular mechanisms are not completely understood, so further research is needed on the mechanisms and causes of aggrecan damage. Here we provide a narrative review based on articles from national and international journals to describe the structure, function, and factors that contribute to the degradation of aggrecan. The results of the study show that changes in the structure of aggrecan are closely related to changes in the mechanical function of joint cartilage. This change occurs mainly due to degradation caused by enzyme activity, a family of matrix metalloproteinase (MMP) and a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS). The present study concludes that aggrecan degradation caused by enzyme activity was very crucial in the development of OA, it was needed to find MMP and ADAMTS inhibitors as a therapeutic agent to prevent the development and progression of OA. Keywords: osteoarthritis, extracellular matrix, aggrecan, degradation

scholarly journals Down-regulation of microRNA-216b inhibits IL-1β-induced chondrocyte injury by up-regulation of Smad3

2017 ◽  
Vol 37 (2) ◽  
Author(s):  
Jiye He ◽  
Jiahong Zhang ◽  
Dongliang Wang
Keyword(s):  
Cell Proliferation ◽  
Type Ii Collagen ◽  
Joint Disease ◽  
Luciferase Reporter ◽  
Type Ii ◽  
Articular Chondrocyte ◽  
Down Regulation ◽  
Protein Levels ◽  
Normal Tissues ◽  
Smad3 Expression

Osteoarthritis (OA) is the most common type of joint disease, leading to a major cause of pain and disability. OA is characterized by the continuous degradation of articular cartilage, mainly resulting in an imbalance between synthesis and degradation of articular chondrocyte extracellular matrix (ECM). Aberrant miR-216b expression has been found in multiple cancers. However, the level of miR-216b in OA cartilage and its role in progression of this disease are still unknown. In the present study, the functional roles of miR-216b and its expression in OA tissues and interleukin-1β (IL-1β)-induced chondrocytes were examined. We found that the level of miR-216b was significantly higher and Smad3 expression was obviously lower in OA cartilage and IL-1β-induced chondrocytes than in normal tissues and cells. Furthermore, a bioinformatics analysis and luciferase reporter assay identified Smad3 as a direct target gene of miR-216b, and Smad3 expression was reduced by miR-216b overexpression at both the mRNA and protein levels. A functional analysis demonstrated that miR-216b down-regulation obviously alleviated the IL-1β-induced inhibition in cell proliferation, type II collagen, and aggrecan down-regulation and matrix metalloproteinase-13 (MMP-13) up-regulation, while miR-216b overexpression had the opposite effects. Knockdown of Smad3 by siRNA reversed the effects of the miR-216b inhibitor on cell proliferation, the expressions of type II collagen, aggrecan, and MMP-13. Our results suggested that miR-216b contributes to progression of OA by directly targeting Smad3, providing a potential therapeutic target for treatment of OA.

scholarly journals LINGO-1 shRNA protects the brain against ischemia/reperfusion injury by inhibiting the activation of NF-κB and JAK2/STAT3

Human Cell ◽  
2021 ◽  
Author(s):  
Jiaying Zhu ◽  
Zhu Zhu ◽  
Yipin Ren ◽  
Yukang Dong ◽  
Yaqi Li ◽  
...  
Keyword(s):  
Cerebral Ischemia ◽  
Nuclear Translocation ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Glucose Deprivation ◽  
Artery Occlusion ◽  
Mice Model ◽  
Down Regulation

AbstractLINGO-1 may be involved in the pathogenesis of cerebral ischemia. However, its biological function and underlying molecular mechanism in cerebral ischemia remain to be further defined. In our study, middle cerebral artery occlusion/reperfusion (MACO/R) mice model and HT22 cell oxygen–glucose deprivation/reperfusion (OGD/R) were established to simulate the pathological process of cerebral ischemia in vivo and in vitro and to detect the relevant mechanism. We found that LINGO-1 mRNA and protein were upregulated in mice and cell models. Down-regulation LINGO-1 improved the neurological symptoms and reduced pathological changes and the infarct size of the mice after MACO/R. In addition, LINGO-1 interference alleviated apoptosis and promoted cell proliferation in HT22 of OGD/R. Moreover, down-regulation of LINGO-1 proved to inhibit nuclear translocation of p-NF-κB and reduce the expression level of p-JAK2 and p-STAT3. In conclusion, our data suggest that shLINGO-1 attenuated ischemic injury by negatively regulating NF-KB and JAK2/STAT3 pathways, highlighting a novel therapeutic target for ischemic stroke.

Down-regulation of chondrocyte aggrecan and type-II Collagen gene expression correlates with increases in static compression magnitude and duration

1999 ◽  
Vol 17 (6) ◽  
pp. 836-842 ◽  
Author(s):  
Paula M. Ragan ◽  
Alison M. Badger ◽  
Michael Cook ◽  
Vicki I. Chin ◽  
Maxine Gowen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Type Ii Collagen ◽  
Collagen Gene ◽  
Type Ii ◽  
Down Regulation ◽  
Static Compression ◽  
Collagen Gene Expression

scholarly journals Prevalence and intensity of chronic pain and self-perceived health among elderly people: a population-based study

2014 ◽  
Vol 22 (4) ◽  
pp. 662-669 ◽  
Author(s):  
Lilian Varanda Pereira ◽  
Patrícia Pereira de Vasconcelos ◽  
Layz Alves Ferreira Souza ◽  
Gilberto de Araújo Pereira ◽  
Adélia Yaeko Kyosen Nakatani ◽  
...  
Keyword(s):  
Chronic Pain ◽  
Elderly People ◽  
Perceived Health ◽  
The Elderly ◽  
The Self ◽  
Lower Limbs ◽  
Joint Disease ◽  
Lumbar Region ◽  
Self Perception ◽  
Perception Of Health

OBJECTIVE: to identify the prevalence and intensity of chronic pain among elderly people of the community and to analyze associations with the self-perceived health status.METHOD: cross-sectional study with a populational sample (n=934), conducted through household interviews in the city of Goiânia, Brazil. The intensity of chronic pain (existing for 6 months or more) was measured using a numerical scale (0-10) and the self-perceived health through a verbal scale (very good, good, fair, poor, very poor). For the statistical analysis, the absolute frequency and percentage, CI (95%), Chi-square test, Odds ratio, and regression analysis were used. Significance of 5%.RESULTS: The prevalence of chronic pain was 52.8% [CI (95%):49.4-56.1]; most frequently located in the lower limbs (34.5%) and lumbar region (29.5%); with high or the worst possible intensity for 54.6% of the elderly people. The occurrence of chronic pain was associated with (p<0.0001) a worse self-perception of health (OR=4.2:2.5-7.0), a greater number of chronic diseases (OR=1.8:1.2-2.7), joint disease (OR=3.5:2.4-5.1) and the female gender (OR=2.3:1.7-3.0). A lower intensity of chronic pain was associated with a better self-perception of health (p<0.0001).CONCLUSION: the majority of the elderly people of the community reported chronic pain, of a severe intensity, and located in areas related to movement activities, thus influencing the morbidity and mortality of this population.

scholarly journals Effects of Extracellular Matrix on the Morphology and Behaviour of Rabbit Auricular Chondrocytes in Culture

2005 ◽  
Vol 2005 (4) ◽  
pp. 364-373 ◽  
Author(s):  
Vega Villar-Suárez ◽  
B. Colaço ◽  
I. Calles-Venal ◽  
I. G. Bravo ◽  
J. G. Fernández-Álvarez ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Type Ii Collagen ◽  
Cell Phenotype ◽  
Nodule Formation ◽  
Type Ii ◽  
Protein Extract ◽  
Positive Effects ◽  
Reticular Pattern ◽  
Chondrocyte Cultures ◽  
Plastic Surfaces

Isolated chondrocytes dedifferentiate to a fibroblast-like shape on plastic substrata and proliferate extensively, but rarely form nodules. However, when dissociation is not complete and some cartilage remnants are included in the culture, proliferation decreases and cells grow in a reticular pattern with numerous nodules, which occasionally form small cartilage-like fragments. In an attempt to reproduce this stable chondrogenic state, we added a cartilage protein extract, a sugar extract, and hyaluronan to the medium of previously dedifferentiated chondrocytes. When protein extract was added, many cartilaginous nodules appeared. Hyaluronan produced changes in cell phenotype and behaviour, but not nodule formation. Protein extract has positive effects on the differentiation of previously proliferated chondrocytes and permits nodule formation and the extensive production of type-II collagen. A comparison with incompletely dissociated chondrocyte cultures suggests that the presence of some living cells anchored to their natural extracellular matrix provides some important additional factors for the phenotypical stability of chondrocytes on plastic surfaces. In order to elucidate if it is possible that the incidence of apoptosis is related to the results, we also characterized the molecular traits of apoptosis.

scholarly journals Store-operated calcium entry suppressed the TGF-β1/Smad3 signaling pathway in glomerular mesangial cells

2017 ◽  
Vol 313 (3) ◽  
pp. F729-F739 ◽  
Author(s):  
Sarika Chaudhari ◽  
Weizu Li ◽  
Yanxia Wang ◽  
Hui Jiang ◽  
Yuhong Ma ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Transforming Growth Factor ◽  
Nuclear Translocation ◽  
Mesangial Cells ◽  
Underlying Mechanism ◽  
Glomerular Mesangial Cells ◽  
Critical Pathway ◽  
Smad3 Phosphorylation ◽  
Tgf Β1

Our previous study demonstrated that the abundance of extracellular matrix proteins was suppressed by store-operated Ca2+entry (SOCE) in mesangial cells (MCs). The present study was conducted to investigate the underlying mechanism focused on the transforming growth factor-β1 (TGF-β1)/Smad3 pathway, a critical pathway for ECM expansion in diabetic kidneys. We hypothesized that SOCE suppressed ECM protein expression by inhibiting this pathway in MCs. In cultured human MCs, we observed that TGF-β1 (5 ng/ml for 15 h) significantly increased Smad3 phosphorylation, as evaluated by immunoblot. However, this response was markedly inhibited by thapsigargin (1 µM), a classical activator of store-operated Ca2+channels. Consistently, both immunocytochemistry and immunoblot showed that TGF-β1 significantly increased nuclear translocation of Smad3, which was prevented by pretreatment with thapsigargin. Importantly, the thapsigargin effect was reversed by lanthanum (La3+; 5 µM) and GSK-7975A (10 µM), both of which are selective blockers of store-operated Ca2+channels. Furthermore, knockdown of Orai1, the pore-forming subunit of the store-operated Ca2+channels, significantly augmented TGF-β1-induced Smad3 phosphorylation. Overexpression of Orai1 augmented the inhibitory effect of thapsigargin on TGF-β1-induced phosphorylation of Smad3. In agreement with the data from cultured MCs, in vivo knockdown of Orai1 specific to MCs using a targeted nanoparticle small interfering RNA delivery system resulted in a marked increase in abundance of phosphorylated Smad3 and in nuclear translocation of Smad3 in the glomerulus of mice. Taken together, our results indicate that SOCE in MCs negatively regulates the TGF-β1/Smad3 signaling pathway.

Differential expression of Rel/NF-κB and octamer factors is a hallmark of the generation and maturation of dendritic cells

Blood ◽  
2000 ◽  
Vol 95 (1) ◽  
pp. 277-285 ◽  
Author(s):  
M. Neumann ◽  
H.-W. Fries ◽  
C. Scheicher ◽  
P. Keikavoussi ◽  
A. Kolb-Mäurer ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Differential Expression ◽  
Antigen Processing ◽  
Nuclear Translocation ◽  
Maturation Stage ◽  
Down Regulation ◽  
Immature Dendritic Cells ◽  
Monocytes And Macrophages ◽  
Induced Changes

Abstract A key feature of maturation of dendritic cells is the down-regulation of antigen-processing and up-regulation of immunostimulatory capacities. To study the differential expression of transcription factors in this process, we investigated the nuclear translocation and DNA binding of Rel/NF-κB and octamer factors during in vitro generation and maturation of dendritic cells compared with macrophage development. RelB was the only factor strongly up-regulated during the generation of both immature dendritic cells and macrophages. Cytokine-induced maturation of dendritic cells resulted in an increase in nuclear RelB, p50, p52, and especially c-Rel, whereas cytokine-treated macrophages responded poorly. This up-regulation of NF-κB factors did not correlate with lower levels of cytosolic NF-κB inhibitors, the IκBs. One IκB, Bcl-3, was strongly expressed only in mature dendritic cells. Furthermore, generation and maturation of dendritic cells led to a continuous down-regulation of the octamer factor Oct-2, whereas monocytes and macrophages displayed high Oct-2 levels. A similar pattern of maturation-induced changes in transcription factor levels was found in cultured murine epidermal Langerhans cells, suggesting a general physiological significance of these findings. Finally, this pattern of differential activation of Rel and octamer factors appears to be suitable in determining the maturation stage of dendritic cells generated by treatment with different cytokine combinations in vitro. (Blood. 2000;95:277-285)

LncRNA MEG3 Inhibits the Degradation of the Extracellular Matrix of Chondrocytes in Osteoarthritis via Targeting miR-93/TGFBR2 Axis

Cartilage ◽  
2019 ◽  
pp. 194760351985575 ◽  
Author(s):  
Kang Chen ◽  
Hao Zhu ◽  
Min-Qian Zheng ◽  
Qi-Rong Dong
Keyword(s):  
Extracellular Matrix ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Cartilage Degradation ◽  
Degenerative Joint Disease ◽  
Joint Disease ◽  
Regulatory Function ◽  
Qrt Pcr ◽  
Ecm Degradation ◽  
Competitive Endogenous Rna

Background As a degenerative joint disease, osteoarthritis (OA) is characterized by articular cartilage degradation. Long noncoding RNAs (lncRNAs) act critical roles in the regulation of OA development, including affecting the proliferation, apoptosis, extracellular matrix (ECM) degradation, and inflammatory response of chondrocytes. The current study’s aim was to investigate the regulatory function and the underlying molecular mechanism of lncRNA MEG3 in ECM degradation of chondrocytes in OA. Methods In the current study, chondrocytes were induced by interleukin-1β (IL-1β) to simulate OA condition, and further assessed cell viability, lncRNA MEG3 and miR-93 expression levels. Overexpression or knockdown of lncRNA MEG3 in chondrocytes treated with IL-1β were performed to investigate the function of MEG3 in regulating cell proliferation, apoptosis and ECM degradation using EdU assay, flow cytometry, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and Western blot. The interaction between MEG3 and miR-93 was assessed using qRT-PCR. Furthermore, overexpression of miR-93 was performed as recovery experiment to explore the functional mechanism of MEG3. Results MEG3 was significantly downregulated in chondrocytes treated with IL-1β, whereas miR-93 was upregulated concomitantly. Overexpression of MEG3 induced the proliferation, suppressed the apoptosis, and relieved the degradation of ECM in IL-1β-induced chondrocytes. By contrast, knockdown of MEG3 suppressed the proliferation, promoted the apoptosis, and aggravated ECM degradation in IL-1β induced chondrocytes. In addition, MEG3 was found to relieve the inhibitive expression of TGFBR2 as a competitive endogenous RNA (ceRNA) of miR-93, and then activated transforming growth factor-β (TGF-β) signaling pathway, regulated chondrocytes ECM degradation in IL-1β induced chondrocytes subsequently. Conclusion LncRNA MEG3 targeted miR-93/TGFBR2 axis, regulated the proliferation, apoptosis and ECM degradation of chondrocytes in OA.

Extracellular matrix modulates enterocyte growth via down regulation of c-jun but is independent of p21 and p27 expression

1998 ◽  
Vol 114 ◽  
pp. A1435
Author(s):  
S.I. Wolpert ◽  
K.M. Lally ◽  
J. Li ◽  
J-Y Wang ◽  
B.L. Bass
Keyword(s):  
Extracellular Matrix ◽  
Down Regulation
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