scholarly journals Resveratrol inhibit interleukin-1β induced inflammatory response in human osteoarthritis chondrocytes through NF-κB signaling pathway

2019 ◽  
Author(s):  
Xiao-Fei Shen ◽  
Yi Cheng ◽  
Qi-Rong Dong ◽  
Min-Qian Zheng
Keyword(s):  
Western Blot ◽  
Signaling Pathway ◽  
Optimal Dose ◽  
Chinese Medicines ◽  
Western Blot Method ◽  
Mrna And Protein Expression ◽  
Osteoarthritis Chondrocytes ◽  
Coumarin Compound ◽  
Oa Chondrocytes ◽  
Anti Inflammatory

Abstract Osteoarthritis (OA) is a common degenerative disease and affect millions of people. Resveratrol is a coumarin compound refined from traditional Chinese medicines with potential anti-inflammatory ability. This study aimed to evaluate protective anti-inflammatory effects of resveratrol in human OA chondrocytes. The chondrocytes were isolated from OA patients. CCK-8 method was used to explore the optimal dose of resveratrol for chondrocyte. Next, we used PCR and Western-blot method to assess the relative mRNA and protein expression in IL-1β group. We further explored relevant mechanism of resveratrol for anti-inflammatory in human OA chondrocytes by immunofluorescence and Western-blot. The results showed that resveratrol blocked IL-1β-stimulated production of NO and PGE2. In addition, resveratrol inhibited the expression of COX-2, iNOs, MMP-1, MMP-3, MMP-13, and increased the levels of aggrecan and collagen-II. Mechanistically, resveratrol suppressed IL-1β-induced IκB-α degradation and NF-κB activation. In conclusion, our results demonstrate that resveratrol inhibits inflammation in OA via the regulation of NF-κB signaling pathway, and suggested that resveratrol may be a potential therapeutic agent for OA.

scholarly journals MEG3 promotes proliferation and inhibits apoptosis in osteoarthritis chondrocytes by miR-361-5p/FOXO1 axis

2019 ◽  
Author(s):  
Anying Wang ◽  
Naixia Hu ◽  
Yefeng Zhang ◽  
Yuanzhen Chen ◽  
Changhui Su ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Cell Apoptosis ◽  
Molecular Mechanisms ◽  
Luciferase Reporter ◽  
Non Coding Rna ◽  
Cartilage Cells ◽  
Osteoarthritis Chondrocytes ◽  
Oa Chondrocytes

Abstract Background: This study aimed to investigate the role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) and related molecular mechanisms, in osteoarthritis (OA). Methods: Cartilage tissues of OA patients and healthy volunteers were isolated and cultured. After transfection with the appropriate construct, chondrocytes were classified into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p mimics groups. qRT-PCR was used to detect the expression of MEG3, miR-361-5p and FOXO1 . Western blot, luciferase reporter assay, RIP, CCK-8, and flow cytometry analysis were performed to reveal the morphology, proliferation, and apoptotic status of cartilage cells. Histological analysis and immunostaining were conducted in the OA rat model. Results: Expression of MEG3 and FOXO1 was significantly decreased in OA compared with the normal group, while the expression of miR-361-5p was increased. MEG3 might serve as a ceRNA of miR-361-5p in OA chondrocytes. Moreover, using western blot analyses and the CCK-8 assay, MEG3 was shown to target miR-361-5p/FOXO1, elevate cell proliferation, and impair cell apoptosis. Functional analysis in vivo showed that MEG3 suppressed degradation of the cartilage matrix. Conclusion: MEG3 can contribute to cell proliferation and inhibit cell apoptosis and degradation of extracellular matrix (ECM) via the miR-361-5p/FOXO1 axis in OA chondrocytes.

scholarly journals Morin Exhibits Anti-Inflammatory Effects on IL-1β-Stimulated Human Osteoarthritis Chondrocytes by Activating the Nrf2 Signaling Pathway

2018 ◽  
Vol 51 (4) ◽  
pp. 1830-1838 ◽  
Author(s):  
Yanlong Qu ◽  
Chunlei Wang ◽  
Ning Liu ◽  
Chengzhe Gao ◽  
Fei Liu
Keyword(s):  
Signaling Pathway ◽  
Inflammatory Activity ◽  
Multifactorial Disease ◽  
Nrf2 Signaling Pathway ◽  
Osteoarthritis Chondrocytes ◽  
Anti Inflammatory

Background/Aims: Osteoarthritis (OA) is a multifactorial disease that is associated with inflammation in joints. The purpose of the present study was to investigate the anti-inflammatory activity and mechanism of morin on human osteoarthritis chondrocytes stimulated by IL-1β. Methods: The levels of NO and PGE2 were measured by the Griess method and ELISA. The levels of MMP1, MMP3, and MMP13 were also measured by ELISA. Results: The results revealed that IL-1β significantly increased the production of NO, PGE2, MMP1, MMP3, and MMP13. Additionally, the increases were significantly attenuated by treatment with morin. Furthermore, IL-1β-induced NF-κB activation was suppressed by morin. In addition, the expression of Nrf2 and HO-1 were increased by morin and knockdown of Nrf2 could prevent the anti-inflammatory effects of morin. Conclusion: In conclusion, this study suggested that morin attenuated IL-1β-induced inflammation by activating the Nrf2 signaling pathway.

scholarly journals MEG3 promotes proliferation and inhibits apoptosis in osteoarthritis chondrocytes by miR-361-5p/FOXO1 axis

2019 ◽  
Author(s):  
Anying Wang ◽  
Naixia Hu ◽  
Yefeng Zhang ◽  
Yuanzhen Chen ◽  
Changhui Su ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Cell Apoptosis ◽  
Molecular Mechanisms ◽  
Normal Group ◽  
Luciferase Reporter ◽  
Non Coding Rna ◽  
Osteoarthritis Chondrocytes ◽  
Oa Chondrocytes

Abstract Background: This study aimed to investigate the role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) and related molecular mechanisms in osteoarthritis (OA). Methods: Patients with OA and patients undergoing thigh amputation were enrolled in OA group and normal group, respectively. Cartilage tissues of all patients were isolated and cultured. After different transfections, chondrocytes were classified into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p mimics groups. The qRT-PCR was used to detect the expression of MEG3, miR-361-5p and FOXO1. Western blot, luciferase reporter assay, RIP, CCK-8 and flow cytometry analysis were performed to reveal the morphology, proliferation and apoptosis of cartilage cells. Histological analysis and immunostaining were conducted in OA rat model. Results: The expression of MEG3 and FOXO1 in OA was significantly decreased while miR-361-5p was increased compared with the normal group. MEG3 might serve as a ceRNA of miR-361-5p in OA chondrocytes. Moreover, the western blot and CCK-8 assay showed that MEG3, targeted miR-361-5p/FOXO1, might elevate cell proliferation and impair cell apoptosis. Functional analysis in vivo showed that MEG3 suppressed the cartilage matrix degradation. Conclusion: Taken together, MEG3 can contribute to cell proliferation, inhibit cell apoptosis and extracellular matrix (ECM) degradation via miR-361-5p/FOXO1 axis in OA chondrocytes.

scholarly journals MEG3 promotes proliferation and inhibits apoptosis in osteoarthritis chondrocytes by miR-361-5p/FOXO1 axis

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Anying Wang ◽  
Naixia Hu ◽  
Yefeng Zhang ◽  
Yuanzhen Chen ◽  
Changhui Su ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Cell Apoptosis ◽  
Molecular Mechanisms ◽  
Luciferase Reporter ◽  
Non Coding Rna ◽  
Cartilage Cells ◽  
Osteoarthritis Chondrocytes ◽  
Oa Chondrocytes

Abstract Background This study aimed to investigate the role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) and related molecular mechanisms, in osteoarthritis (OA). Methods Cartilage tissues of OA patients and healthy volunteers were isolated and cultured. After transfection with the appropriate constructs, chondrocytes were classified into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p mimics groups. qRT-PCR was used to detect the expression of MEG3, miR-361-5p and FOXO1. Western blot, luciferase reporter assay, RIP, CCK-8, and flow cytometry analysis were performed to reveal the morphology, proliferation, and apoptotic status of cartilage cells. Histological analysis and immunostaining were conducted in the OA rat model. Results Expression of MEG3 and FOXO1 was significantly decreased in OA compared with the normal group, while the expression of miR-361-5p was increased. MEG3 might serve as a ceRNA of miR-361-5p in OA chondrocytes. Moreover, using western blot analyses and the CCK-8 assay, MEG3 was shown to target miR-361-5p/FOXO1, elevate cell proliferation, and impair cell apoptosis. Functional analysis in vivo showed that MEG3 suppressed degradation of the cartilage matrix. Conclusion MEG3 can contribute to cell proliferation and inhibit cell apoptosis and degradation of extracellular matrix (ECM) via the miR-361-5p/FOXO1 axis in OA chondrocytes.

scholarly journals Andrographolide Inhibits Inflammatory Cytokines Secretion in LPS-Stimulated RAW264.7 Cells through Suppression of NF-κB/MAPK Signaling Pathway

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Yu Li ◽  
Shengnan He ◽  
Jishun Tang ◽  
Nana Ding ◽  
Xiaoyan Chu ◽  
...  
Keyword(s):  
Western Blot ◽  
Signaling Pathway ◽  
Proinflammatory Cytokines ◽  
Molecular Mechanisms ◽  
Mapk Signaling ◽  
Mapk Signaling Pathway ◽  
Raw264.7 Cells ◽  
Nuclear Level ◽  
Expression Levels ◽  
Anti Inflammatory

Andrographolide, the main active component extracted from Andrographis paniculata (Burm.f.) Wall. ex Nees, exerts anti-inflammatory effects; however, the principal molecular mechanisms remain unclear. The objective of this study was to investigate the molecular mechanisms of Andrographolide in modifying lipopolysaccharide- (LPS-) induced signaling pathway in RAW264.7 cells. An in vitro model of inflammation was induced by LPS in mouse RAW264.7 cells in the presence of Andrographolide. The concentration and expression levels of proinflammatory cytokines were determined by an enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. The nuclear level of NF-κB was measured by an electrophoretic mobility shift assay (EMSA). The expression levels of NF-κB, p38, ERK, and JNK were determined by western blot. Andrographolide dose-dependently inhibited the release and mRNA expression of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 cells. The nuclear level of p65 protein was decreased in Andrographolide treatment group. Western blot analysis showed that Andrographolide suppressed LPS-induced NF-κB activation and the phosphorylation of IkBa, ERK1/2, JNK, and p38. These results suggest that Andrographolide exerts an anti-inflammatory effect by inhibiting the activation of NF-κB/MAPK signaling pathway and the induction of proinflammatory cytokines.

scholarly journals MEG3 promotes proliferation and inhibits apoptosis in osteoarthritis chondrocytes by miR-361-5p/FOXO1 axis

2019 ◽  
Author(s):  
Anying Wang ◽  
Naixia Hu ◽  
Yefeng Zhang ◽  
Yuanzhen Chen ◽  
Changhui Su ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Rat Model ◽  
Cell Apoptosis ◽  
Molecular Mechanisms ◽  
Normal Group ◽  
Luciferase Reporter ◽  
Non Coding Rna ◽  
Osteoarthritis Chondrocytes ◽  
Oa Chondrocytes

Abstract Background: This study aimed to investigate the role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) and related molecular mechanisms in osteoarthritis (OA). Methods: Patients with OA and patients undergoing thigh amputation were involved in OA group and normal group, respectively. Cartilage tissues of all patients were isolated and cultured. Based on different transfection, MEG3 cells were grouped into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p mimics group. The RT-qPCR was used to detect the expression of MEG3, miR-361-5p and FOXO1. Moreover, western blot, luciferase reporter assay, RIP, CCK-8 and flow cytometry analysis were performed to reveal the morphology, proliferation and apoptosis in cartilage cells. Finally, the histological analysis and immunostaining were performed on OA rat model. Results: The expression of MEG3 and FOXO1 in OA was significantly decreased while miR-361-5p was increased compared with the normal group. MEG3 might serve as a ceRNA of miR-361-5p in OA chondrocytes. Moreover, the western blot and CCK-8 assay showed that MEG3, targeted miR-361-5p/FOXO1, might elevate cell proliferation and impair cell apoptosis. Finally, rat model analysis showed that MEG3 suppressed the cartilage matrix degradation. Conclusion: Taken together, MEG3 can contribute to cell proliferation, inhibit cell apoptosis and extracellular matrix (ECM) degradation via miR-361-5p/FOXO1 axis in OA chondrocytes.

scholarly journals Study on the mechanism of multi-AGC kinase AT13148 on Notch signaling pathway in glioblastoma

2021 ◽  
Author(s):  
Yanan Li ◽  
Guosheng Han ◽  
Weijie Min ◽  
Mengmeng Li ◽  
Maomao Wang ◽  
...  
Keyword(s):  
Western Blot ◽  
Notch Signaling ◽  
Signaling Pathway ◽  
Small Molecule ◽  
Notch Signaling Pathway ◽  
Small Molecule Drug ◽  
Mrna And Protein Expression ◽  
U87 Cells ◽  
Proliferation And Invasion

IntroductionGlioblastoma is the most malignant astrocytoma, and its therapeutic effect is not ideal. Notch signaling pathway plays an important role in tumor proliferation and invasion. Whether small molecule drug AT13148 can affect glioblastoma by regulating Notch signaling pathway is the focus of this study.Material and methodsIn vitro, glioblastoma U87 cell line transfected with sh-ITGB1 (U87sh-ITGB1), U87 cell line transfected with oe-ITGB1 (U87oe-ITGB1) and control group were treated with a small molecular drug AT13148. RT-qPCR, western-blot and clone formation ability assays were used to detect the mRNA and protein expression of the ITGB1 and the key gene NOTCH1, as well as the proliferation of cancer cells. Therapeutic effects of AT13148 were examined in vivo using a nude mice model of U87 cells. After treatment with AT13148, volume of tumors were calculated, and RT-qPCR and western-blot were used to evaluate the mRNA and protein expression of the ITGB1 and NOTCH1.ResultsAT13148 inhibits the activity of U87 cells. Lentiviral transfection of sh-ITGB1 and oe-ITGB1 can interfere with the expression of ITGB1 in U87 cells. AT13148 could down-regulate both the expression of ITGB1 and NOTCH1. Moreover, AT13148 affects the cloning ability of U87 cells. AT13148 can also inhibit the proliferation of U87 cells. Furthermore, AT13148 inhibited the proliferation and invasion of transplanted tumors in vivo.ConclusionsThis study indicated that AT13148 could affect the expression of ITGB1 and NOTCH1, which also could be a potential potential anti-glioblastoma small molecule drug candidate in clinic medicine.

scholarly journals Activation of Hippocampal IR/IRS-1 Signaling Contributes to the Treatment with Zuogui Jiangtang Jieyu Decoction on the Diabetes-Related Depression

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Hui Yang ◽  
Jia Ling ◽  
Pan Meng ◽  
Jian Liu ◽  
Xiaoyuan Lin ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Blood Glucose ◽  
Western Blot ◽  
Insulin Receptor ◽  
Signaling Pathway ◽  
Diabetic Rats ◽  
Morris Water Maze Test ◽  
Western Blot Method ◽  
Peripheral Insulin Resistance ◽  
The Brain

Background. Zuogui Jiangtang Jieyu decoction (ZJJ) is mainly used for the treatment of diabetes-related depression in current clinical applications and research. This study aims to investigate whether the brain IR/IRS-1 signaling pathway is involved in the therapeutic effect of ZJJ on depression-like behavior in diabetic rats. Methods. Sprague–Dawley rats were fed with high-fat diet and subjected to streptozotocin injection to establish the diabetes animal model. After treatment with different doses of ZJJ (20.530 g/kg or 10.265 g/kg) for 4 weeks, the blood glucose level and peripheral insulin resistance were measured. The forced-swimming test (FST) and Morris water maze test (MWMT) were applied for the mood and cognitive function assessment. Then, the Western blot method was used to analyze the protein levels of insulin receptor (IR), insulin receptor substrate-1 (IRS-1), phosphatidylonositol-3-kinase (PI3K), and protein kinase B (PKB, also as known as AKT) in the hippocampus of diabetic rats. Meanwhile, the immunofluorescence method was performed to analyze the above proteins’ expression in the neuron and astrocyte. At last, the levels of glycogen, lactate, and ATP were tested by the ELISA method. Additionally, the insulin-sensitive glucose transporter 4 (GLUT4) and the lactate transporter monocarboxylate transporter 4 (MCT4) were analyzed by the Western blot method. Results. ZJJ administration significantly decreased the level of blood glucose and improved the peripheral insulin resistance in diabetic rats. Besides, ZJJ attenuated the depression-like behavior and the cognitive dysfunction in rats with diabetes. Furthermore, we found the upregulation of protein expression of phospho-IR, phospho-IRS-1, phospho-PI3K, and phospho-AKT in the hippocampus of diabetic rats after being treated with ZJJ. Moreover, the above proteins are increased not only in the neuron but also in the astrocyte after ZJJ administration. In addition, ZJJ increased the content of ATP, glycogen, and lactate, as well as the expression of GLUT4 and MCT4 in the hippocampus of diabetic rats. Conclusions. These findings suggest that ZJJ improves the depression-like behavior of diabetic rats by activating the IR/IRS-1 signaling pathway in both hippocampal neuron and astrocyte. And the brain IR/IRS-1 signaling pathway plays an important role in astrocyte-neuron metabolic coupling, providing a potential mechanism by which the IR/IRS-1 signaling pathway may contribute to the treatment of ZJJ on diabetes-related depression.

scholarly journals Bergenin-activated SIRT1 inhibits TNF-a-induced pro-inflammatory response via blocking NF-kB signaling pathway 

2020 ◽  
Author(s):  
Bei xian Zhou ◽  
Bin Wu ◽  
Min Chen ◽  
Cuifen Chen ◽  
Yun Gao ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Signaling Pathway ◽  
Western Blotting ◽  
Nuclear Translocation ◽  
Sirtuin 1 ◽  
Burn Wound ◽  
Burn Wound Healing ◽  
Mrna And Protein Expression ◽  
Sirt1 Inhibitor ◽  
Anti Inflammatory

Abstract BackgroundBergenin, a kind of polyphenol compound, has been showed to exhibit antiulcerogenic, anti-inflammatory, antitussive and burn wound healing properties, However, its therapeutic effect on TNF-a induced pro-inflammatory response in airway and its potential mechanisms of actions were still unclearAim of this studyThis study aimed to investigate the anti-inflammatory effects and possible mechanism of bergenin in TNF-a-stimulated 16-HBE cells.Materials and methodsCCK8 was used to determine the cytotoxicity. Cytokines expression were analyzed by qRT-qPCR and ELISA. Immunofluorescence, western blotting and SIRT1 activity assay were employed to investigate the possible molecular mechanism. ResultsOur results showed that bergenin treatment obviously decreased the mRNA and protein expressions of IL-6 and IL-8 in TNF-a-stimulated 16-HBE cells. Bergenin blocked TNF-a-mediated the activation of NF-kB signaling and NF-kB nuclear translocation. Interestingly, RT‑qPCR and western blotting revealed that bergenin did not affect the expression of sirtuin-1 (SIRT1), but significantly increased the activity of SIRT1. Furthermore, bergenin-mediated SIRT1 activation was further confirmed by the resulted that bergenin decreased acetylation levels of NF-kB-p65 and p53. The inhibitory effects of bergenin on the mRNA and protein expression levels of IL-6 and IL-8 were reversed by addition of SIRT1 inhibitor. In addition, combination of bergenin and dexamethasone showed additive effects on the reduction of IL-6 and IL-8.ConclusionsThese findings demonstrated that bergenin could suppress TNF-α-induced pro-inflammatory response via augmenting SIRT1 activity to block NF-kB signaling pathway, which may contribute to provide beneficial effect for the treatment of airway inflammation in asthma.

scholarly journals High expression of an unknown long noncoding RNA RP11-290L1.3 from GDM macrosomia and its effect on preadipocyte differentiation

2021 ◽  
Author(s):  
Yu Lin ◽  
Yingying Zhang ◽  
Lei Xu ◽  
Wei Long ◽  
Chunjian Shan ◽  
...  
Keyword(s):  
Protein Expression ◽  
Western Blot ◽  
Signaling Pathway ◽  
Pathway Analysis ◽  
Long Noncoding Rna ◽  
Noncoding Rna ◽  
Adipocyte Differentiation ◽  
Preadipocyte Differentiation ◽  
Expression Levels ◽  
Mrna And Protein Expression

Aims: Gestational diabetes mellitus (GDM)-induced macrosomia is predominantly characterized by fat accumulation, which is closely related to adipocyte differentiation. An unknown long noncoding RNA RP11-290L1.3, referred to as RP11, was identified to be dramatically upregulated in the umbilical cord blood of women with GDM-induced macrosomia in our previous study. We conducted this study to identify the function of RP11 in GDM-induced macrosomia. Methods: The effects of RP11 gain- and loss-of-function on HPA-v (human preadipocytes-visceral) adipogenesis were determined with lentivirus mediated cell transduction. The mRNA and protein expression levels of adipogenesis makers were evaluated by qPCR/western blot. Then, we performed the Microarray and pathway analysis to explore the possible mechanisms by which RP11 regulates adipogenesis. Results: Overexpression of RP11 significantly enhanced adipocyte differentiation and increased the mRNA and protein expression levels of adipogenesis makers, such as PPAR-γ, SREBP1c, and FASN by qPCR/western blot. Knockdown of RP11 showed opposite effects. Microarray and pathway analysis showed, after RP11 knockdown, 1,612 genes were upregulated and 583 genes were downregulated which were found to be mainly involved in metabolic pathways, insulin signaling pathway and MAPK signaling pathway. Conclusion: In conclusion, the unknown lncRNA RP11 serves a positive factor on preadipocyte differentiation which could shed light on fetal fat accumulation in GDM.

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