scholarly journals LncRNA cardiac autophagy inhibitory factor is downregulated in rheumatoid arthritis and suppresses the apoptosis of fibroblast-like synoviocytes by promoting the maturation of miRNA-20a

2021 ◽  
Author(s):  
Dongming Xu ◽  
Ling Lin ◽  
Zhen Chen
Keyword(s):  
Rheumatoid Arthritis ◽  
Cell Apoptosis ◽  
Quantitative Polymerase Chain Reaction ◽  
Linear Regression Analysis ◽  
Inhibitory Factor ◽  
Dependent Manner ◽  
Plasma Samples ◽  
Expression Levels ◽  
Apoptosis Assay ◽  
Fibroblast Like Synoviocytes

Objectives: In this study, we aimed to investigate the effects of LncRNA cardiac autophagy inhibitory factor (CAIF) and miR-20a on the apoptosis of synovial cells in rheumatoid arthritis (RA) and the regulatory mechanism. Patients and methods: Between May 2018 and March 2020, a total of 62 RA patients (24 males, 38 females; mean age: 55.2±4.9 years; range, 42 to 68 years) and 62 controls (24 males, 38 females; mean age: 55.3±4.8 years; range, 41 to 68 years) were included in this study. Plasma samples were collected from all participants. The expression levels of CAIF, mature miR-20a, and miR-20a precursor in these plasma samples were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Correlations were analyzed using linear regression analysis. Overexpression of CAIF was achieved in human fibroblast-like synoviocytes (HFLSs) and the expression levels of mature miR-20a and miR-20a precursor were determined using RT-qPCR. Cell apoptosis was analyzed by cell apoptosis assay. Results: The CAIF was downregulated in RA and positively correlated with the expression of mature miR-20a. In HFLSs, LPS treatment resulted in downregulation of both CAIF and miR-20a in a dose-dependent manner. In HFLSs, overexpression of CAIF did not affect the expression of miR-20a precursor, but upregulated the expression of mature miR-20a. Cell apoptosis analysis showed that overexpression of CAIF and miR-20a inhibited the apoptosis of HFLSs induced by LPS. The combination of overexpression of CAIF and miR-20a showed a stronger effect. Conclusion: The CAIF may suppress the apoptosis of HFLSs in RA by promoting the maturation of miR-20a.

scholarly journals Expression of AIM2 in rheumatoid arthritis and its role on fibroblast-like synoviocyte

2020 ◽  
Author(s):  
fujuan qiu ◽  
Chen Yong ◽  
Qiu Fujuan ◽  
Zhao Xiaofeng ◽  
Xiao Changhong
Keyword(s):  
Rheumatoid Arthritis ◽  
Mtt Assay ◽  
Control Group ◽  
Expression Levels ◽  
Potential Target ◽  
Caspase 1 ◽  
Significant Difference ◽  
And Migration ◽  
Fibroblast Like Synoviocytes ◽  
Proliferation And Migration

Abstract Background To determine whether any differences of AIM2 inflammasome expression levels between rheumatoid arthritis (RA) and osteoarthritis (OA) and investigate the effects of AIM2 when transferred into RA fibroblast-like synoviocytes (RA-FLS).Methods Serum AIM2 levels between OA and RA patients were compared by ELISA. Different expression levels of AIM2, ASC, Caspase-1 and IL-1β between RA and OA synovium were semi-quantified by RT-qPCR and immunohistochemical (IHC) staining. IHC staining were recorded by H scores, and determine the correlation with ESR and CRP levels of RA patients. SiRNA AIM2 was transferred to RA-FLS and observe its effects on proliferation and migration by MTT assay and transwell test respectively.Results In RA sera, no significant difference was observed between OA and RA patients. However, in affected knee synovium, AIM2, ASC, Caspase-1 and IL-1β were expressed higher in RA than that of OA. Plus, H score of AIM2, ASC, and IL-1β were positively correlated to ESR and CRP levels in RA patients. After transferred AIM2 siRNA to FLS and incubation for 48 hours, the proliferation of FLS were significantly inhibited, and the apoptosis rate were significantly increased compared to FLS in control group. However, no effect on migration was detected.Conclusions AIM2 participated in the proliferation of FLS, and might be a potential target for therapy.

scholarly journals Protective Effects of Pretreatment with Quercetin Against Lipopolysaccharide-Induced Apoptosis and the Inhibition of Osteoblast Differentiation via the MAPK and Wnt/β-Catenin Pathways in MC3T3-E1 Cells

2017 ◽  
Vol 43 (4) ◽  
pp. 1547-1561 ◽  
Author(s):  
Chun Guo ◽  
Rui-Juan Yang ◽  
Ke Jang ◽  
Xiao-ling Zhou ◽  
Yu-zhen Liu
Keyword(s):  
Cytochrome C ◽  
Osteoblast Differentiation ◽  
Caspase 3 ◽  
Quantitative Polymerase Chain Reaction ◽  
Bone Diseases ◽  
Cell Counting ◽  
Dependent Manner ◽  
Protective Effects ◽  
Expression Levels ◽  
Induced Apoptosis

Background/Aims: Quercetin, a flavonoid found in onions and other vegetables, has potential inhibitory effects on bone resorption in vivo and in vitro. In our previous study, we found that quercetin treatment reversed lipopolysaccharide (LPS)-induced inhibition of osteoblast differentiation through the mitogen-activated protein kinase (MAPK) pathway in MC3T3-E1 cells. In this study, we investigated the underlying mechanisms of pretreatment with quercetin on apoptosis and the inhibition of osteoblast differentiation in MC3T3-E1 cells induced by LPS. Methods: MC3T3-E1 osteoblasts were treated with quercetin for 2 h; cells were then incubated with LPS in the presence of quercetin for the indicated times. Cell viability was measured using the Cell Counting Kit-8 (CCK-8) assay, and cell apoptosis was evaluated using Hoechst 33258 staining. The mRNA expression levels of osteoblast-specific genes, Bax and caspase-3 were determined by real-time quantitative polymerase chain reaction (qPCR). Protein levels of osteoblast-specific genes, caspase-3, Bax, cytochrome c, Bcl-2, Bcl-XL, phosphorylated MAPKs and Wnt/β-catenin were measured using Western blot assays. The MAPK and Wnt/β-catenin signalling pathways were blocked prior to pretreatment with quercetin. Results: Pretreatment with quercetin significantly restored LPS-suppressed bone mineralization and the mRNA and protein expression levels of osteoblast-specific genes such as Osterix (OSX), runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteocalcin (OCN) in a dose-dependent manner. Pretreatment with quercetin also inhibited osteoblast apoptosis, significantly restored the down-regulated expression of Bcl-2 and Bcl-XL and decreased the upregulated expression of caspase-3, Bax, and cytochrome c in MC3T3-E1 cells induced by LPS. Furthermore, pretreatment with quercetin not only decreased the abundance of phosphorylated p38 MAPK and increased the abundance of phosphorylated extracellular signal regulated kinase (ERK), but also triggered the Wnt/β-catenin pathway through enhancing expression of Wnt3 and β-catenin. Pretreatment with MAPK inhibitors or the Wnt/β-catenin inhibitor XAV939 blocked the protective effects of quercetin against LPS-induced apoptosis and the inhibition of osteoblast differentiation. Conclusions: Our findings suggest that pretreatment with quercetin may be a potential drug for preventing abnormal human bone loss induced by LPS in bacteria-induced bone diseases.

scholarly journals MiR-498 suppresses proliferation and inflammation in fibroblast-like synoviocytes in rheumatoid arthritis via targeting JAK1

2021 ◽  
Vol 18 (10) ◽  
pp. 2011-2017
Author(s):  
Lan Chai ◽  
Xian Zhen Zhang ◽  
Hai fang Ma ◽  
Fang Yuan
Keyword(s):  
Rheumatoid Arthritis ◽  
Cell Cycle ◽  
Polymerase Chain Reaction ◽  
Therapeutic Target ◽  
Cell Apoptosis ◽  
Inflammatory Responses ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Synovial Tissues ◽  
Fibroblast Like Synoviocytes

Purpose: To investigate the effect of microRNA 498 (miR-498) on proliferation and inflammation of rheumatoid arthritis (RA) fibroblast-like synoviocytes (RA-FLSs) in rheumatoid arthritis (RA). Methods: MiR-498 level was evaluated in both RA synovial tissues and RA-FLSs using real-time polymerase chain reaction (PCR). MicroRNA-498 overexpression or knockdown was performed in RAFLSs. Proliferation, apoptosis, cell cycle and inflammation induced by miR-498 mimics or inhibitor were used to explore the function of miR-498 in RA. Results: Expression level of miR-498 was downregulated in both RA synovial tissues and RA- FLSs. MicroRNA-498 mimics decreased proliferation and arrested cell cycle, whereas miR-498 inhibitor caused the opposite effects in RA-FLSs. In addition, miR-498 mimics suppressed inflammation and promoted cell apoptosis, while miR-498 inhibitor promoted inflammation and inhibited cell apoptosis in RA-FLSs. Furthermore, the effect of miR-498 on the proliferation, inflammation and apoptosis of RAFLSs was mediated by its ability to target and downregulate JAK1. Conclusion: These results indicate that miR-498 inhibits the proliferation and inflammatory responses of RA-FLSs by targeting JAK1, thus revealing a new therapeutic target for RA treatment.

scholarly journals LncRNA MALAT1 Forms a Negative Feedback Loop With lncRNA CRNDE in Sepsis to Regulate Lung Cell Apoptosis

2020 ◽  
Author(s):  
Caifang Yue ◽  
Muhan He ◽  
Yanping Teng ◽  
Xiaoli Bian
Keyword(s):  
Negative Feedback ◽  
Cell Apoptosis ◽  
Feedback Loop ◽  
Potential Interaction ◽  
Negative Feedback Loop ◽  
Lung Cells ◽  
Plasma Samples ◽  
Lncrna Malat1 ◽  
Apoptosis Assay ◽  
The Relationship

Abstract Background: It has been reported that lncRNAs MALAT1 and CRNDE plays opposite roles in sepsis. Therefore, they may have crosstalk. We therefore explored the potential interaction between them in sepsis. Methods: Plasma samples were collected from both sepsis patients (n=60) and healthy controls (n=60). Expression of MALAT1 and CRNDE in plasma samples before than after treatment was determined by RT-qPCR. Results: Overexpression of MALAT1 and CRNDE in Human Bronchial Epithelial Cells (HBEpCs) was achieved to explore the relationship between them. The roles of MALAT1 and CRNDE in regulating the apoptosis of HBEpCs were analyzed by cell apoptosis assay. MALAT1 was upregulated in sepsis, while CRNDE was downregulated in sepsis. In lung cells, overexpression of MALAT1 and CRNDE mediated the downregulation of each other. With proper treatment, MALAT1 was downregulated in sepsis and CRNDE was upregulated in sepsis. In lung cells, LPS mediated the upregulation of MALAT1 and the downregulation of CRNDE. Cell apoptosis analysis showed that MALAT1 overexpression promoted the apoptosis of lung cells induced by LPS, and the overexpression of CRNDE played an opposite role. Moreover, CRNDE overexpression attenuated the effects of MALAT1 overexpression. Conclusion: MALAT1 may form a negative feedback loop with CRNDE in sepsis to regulate lung cell apoptosis.

scholarly journals Circ_0038467 is Overexpressed in Osteoarthritis and it Promotes the Maturation of miR-203 to Increase the Apoptosis of Chondrocytes Induced by LPS

2021 ◽  
Author(s):  
Zhongkun Gou ◽  
Quanling Wu ◽  
Changqing Jiang ◽  
Wei Dong
Keyword(s):  
Cell Apoptosis ◽  
Critical Role ◽  
Expression Levels ◽  
Apoptosis Assay ◽  
Lps Treatment

Abstract Introduction: LPS-induced inflammation contributes to osteoarthritis (OA). It is known that Circ_0038467 plays a critical role in LPS-mediated inflammation, suggesting the its involvement in OA. This study aimed to study the possible involvement of Circ_0038467 in OA.Materials and Methods: Circ_0038467, mature miR-203 and miR-203 precursor expression in controls and OA patients was determined by RT-qPCR. Circ_0038467 overexpression in chondrocytes and RT-qPCR was performed to analyze its effects on the expression of mature miR-203 and miR-203 precursor. The role of Circ_0038467 and miR-203 in cell apoptosis was analyzed by cell apoptosis assay.Results: Circ_0038467 was upregulated in OA and positively correlated with mature miR-203, but not miR-203 precursor. In chondrocytes, increased expression of both Circ_0038467 and miR-203 was observed after LPS treatment. In chondrocytes, Circ_0038467 overexpression increased the expression levels of mature miR-203, but not miR-203 precursor. Analysis of cell apoptosis showed that overexpression of Circ_0038467 and miR-203 increased cell apoptosis. In addition, miR-203 inhibitor reversed the effects of Circ_0038467 overexpression on cell apoptosis.Conclusions: Circ_0038467 is highly expressed in OA and may promote the production of mature miR-203 to increase the apoptosis of chondrocytes induced by LPS.

scholarly journals Proinflammatory Effects of Ubiquitin-Specific Protease 5 (USP5) in Rheumatoid Arthritis Fibroblast-Like Synoviocytes

2020 ◽  
Vol 2020 ◽  
pp. 1-9 ◽  
Author(s):  
Xiao-Bo Luo ◽  
Jian-Cheng Xi ◽  
Zhen Liu ◽  
Yu Long ◽  
Li-tao Li ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Tumor Necrosis Factor Receptor ◽  
Dependent Manner ◽  
Autoimmune Inflammatory Disease ◽  
Specific Protease ◽  
Inflammatory Processes ◽  
Ubiquitin Specific Protease ◽  
Signaling Activation ◽  
Fibroblast Like Synoviocytes

Rheumatoid arthritis (RA) is a worldwide chronic autoimmune inflammatory disease which is affecting approximately 1% of the total population. It is characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS) and increased production of proinflammatory cytokines. In the current study, we were aiming to investigate the role of ubiquitin-specific protease 5 (USP5) in the inflammatory process in RA-FLS. Expression of USP5 was found upregulated in RA-FLS compared with that in osteoarthritis- (OA-) FLS, and IL-1β stimulation increased USP5 expression in a time-dependent manner. Furthermore, we found that USP5 overexpression significantly aggravated proinflammatory cytokine production and related nuclear factor κB (NF-κB) signaling activation. Consistently, silencing of USP5 decreased the release of cytokines and inhibited the activation of NF-κB. In addition, USP5 was found to interact with tumor necrosis factor receptor-associated factor 6 (TRAF6) and remove its K48-linked polyubiquitination chains therefore stabilizing TRAF6. Our data showed that a USP5-positive cell regulates inflammatory processes in RA-FLS and suggested USP5 as a potential target for RA treatment.

Overexpression of lncRNA MCM3AP-AS1 in Sepsis Suppresses the Maturation of miR-223 to Promote LPS-induced Lung Cells

2020 ◽  
Author(s):  
Chunzhi Gong ◽  
Jianying Wang ◽  
Shupeng Liu ◽  
Zunfeng Li ◽  
Zhaoguo Liu ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Lung Cells ◽  
Healthy Controls ◽  
Plasma Samples ◽  
Inhibitory Effects ◽  
Human Bronchial Epithelial Cells ◽  
Bronchial Epithelial ◽  
Apoptosis Assay ◽  
Induced Apoptosis ◽  
Lps Treatment

Abstract Background: Previous studies have showed that lncRNA MCM3AP-AS1 and miR-223 play opposite roles in LPS-induced inflammation, which contributes to the progression of sepsis. This study was therefore carried out to analyze the interactions between MCM3AP-AS1 and miR-223 in sepsis. Methods: Plasma samples were obtained from 62 sepsis patients and 62 healthy controls. Expression of MCM3AP-AS1, miR-223 precursor and mature miR-223 in plasma samples was determined by RT-qPCR. In human bronchial epithelial cells (HBEpCs), the interaction between MCM3AP-AS1 and miR-223 was analyzed by overexpression experiments. Cell apoptosis assay was analyzed by cell apoptosis assay.Results: We found that MCM3AP-AS1 was upregulated in sepsis, while miR-223 was downregulated in sepsis. MCM3AP-AS1 and mature miR-223 were inversely correlated, while MCM3AP-AS1 and miR-223 precursor were not. In HBEpCs, LPS treatment resulted in the upregulation of MCM3AP-AS1 and downregulation of miR-223. In HBEpCs, MCM3AP-AS1 overexpression downregulated mature miR-223 but failed to affect miR-223 precursor. In addition, MCM3AP-AS1 overexpression reduced the inhibitory effects of miR-223 on LPS-induced apoptosis of HBEpCs.Conclusions: LncRNA MCM3AP-AS1 is upregulated in sepsis and may suppress the maturation of miR-223 to promote LPS-induced lung cells.

scholarly journals Circular RNA circFADS2 is overexpressed in sepsis and suppresses LPS-induced lung cell apoptosis by inhibiting the maturation of miR-15a-5p

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Xiaoyang Hong ◽  
Shuanglei Li ◽  
Jie Wang ◽  
Zhe Zhao ◽  
Zhichun Feng
Keyword(s):  
Cell Apoptosis ◽  
Expression Vector ◽  
Circular Rna ◽  
Lung Cells ◽  
Healthy Controls ◽  
Plasma Samples ◽  
Apoptosis Assay ◽  
Induced Apoptosis ◽  
Lps Treatment

Abstract Background Circular RNA circFADS2 plays protective roles in LPS-induced inflammation, which promotes sepsis, suggesting its involvement in sepsis. Methods Expression of circFADS2, mature miR-15a-5p, and miR-15a-5p precursor in plasma samples from sepsis patients and healthy controls was determined by RT-qPCR. The circFADS2 expression vector was transfected in lung cells, followed by the measurement of the expression levels of mature miR-15a-5p and miR-15a-5p precursor to study the role of circFADS2 in miR-15a-5p maturation. Cell apoptosis was analyzed by cell apoptosis assay. Results CircFADS2 was upregulated in sepsis and inversely correlated with mature miR-15a-5p, but not miR-15a-5p precursor. In lung cells, circFADS2 overexpression decreased the level of mature miR-15a-5p, but not miR-15a-5p precursor. LPS treatment decreased miR-15a-5p expression and increased circFADS2 level. Cell apoptosis analysis showed that circFADS2 overexpression reduced miR-15a-5p overexpression-induced apoptosis of LPS-treated lung cells. Conclusions CircFADS2 is upregulated in sepsis to suppress LPS-induced lung cell apoptosis by inhibiting miR-15a-5p maturation.

scholarly journals Identification of drug targets and potential molecular mechanisms for Wantong Jingu Tablet extract in treatment of rheumatoid arthritis: bioinformatics analysis of fibroblast-like synoviocytes

2020 ◽  
Author(s):  
Zhaodong Li ◽  
Fangyuan Qi ◽  
Fan Li
Keyword(s):  
Rheumatoid Arthritis ◽  
Western Blotting ◽  
Drug Targets ◽  
High Performance ◽  
Molecular Mechanisms ◽  
High Throughput Sequencing ◽  
Bioinformatics Analysis ◽  
Underlying Mechanism ◽  
Dependent Manner ◽  
Fibroblast Like Synoviocytes

Abstract Background: Rheumatoid arthritis- fibroblast-like synoviocytes (RA-FLSs) play important roles in pathogenesis of rheumatoid arthritis (RA). Wantong Jingu Tablet (WJT), a mixture of traditional Chinese medicine, is a potentially effective therapy for RA, but its underlying mechanism is unclear. In this study, we explore the effects of WJT on human RA-FLSs and the underlying molecular mechanism. Methods: The major components of WJT were determined using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Cell proliferative ability was evaluated by CCK-8, colony formation assay, and EdU incorporation assay. Cell apoptotic capacity was examined by caspase-3 and caspase-9 activity test. Protein levels of Bax and Bcl-2 were investigated by western blotting. High-throughput sequencing and bioinformatics analysis were conducted to screen and identify targeted genes, followed by identification by qRT-PCR and western blotting. Results: In this study, we have identified 346 compounds in WJT. Our results showed that WJT inhibited the RA-FLSs proliferation, and promoted apoptosis in a dose- and time-dependent manner. More importantly, 184 differentially expressed genes (DEGs) has been screened after WJT treatment based on DEGSeq2 and 278 DEGs was identified by DEGSeq2 combined with WGCNA. Then, 10 hub genes were identified based on two different analyses, while the expression levels of only SMC3, THOC1, BUB1, and STAG2 were decreased after WJT treatment, which was identical to the sequencing profiles.Conclusions: WJT exerted its anti-proliferation and pro-apoptosis effects possibly through suppressing the expression of SMC3, THOC1, BUB1, and STAG2 in RA-FLSs. Thus, therapeutics targeting these genes may be a promising strategy for rescuing RA.

scholarly journals Identification of drug targets and potential molecular mechanisms for Wantong Jingu Tablet extract in treatment of rheumatoid arthritis: bioinformatics analysis of fibroblast-like synoviocytes

2020 ◽  
Author(s):  
Zhaodong Li ◽  
Fangyuan Qi ◽  
Fan Li
Keyword(s):  
Rheumatoid Arthritis ◽  
Western Blotting ◽  
Drug Targets ◽  
High Performance ◽  
Molecular Mechanisms ◽  
High Throughput Sequencing ◽  
Bioinformatics Analysis ◽  
Underlying Mechanism ◽  
Dependent Manner ◽  
Fibroblast Like Synoviocytes

Abstract Background: Rheumatoid arthritis- fibroblast-like synoviocytes (RA-FLSs) play important roles in pathogenesis of rheumatoid arthritis (RA). Wantong Jingu Tablet (WJT), a mixture of traditional Chinese medicine, is a potentially effective therapy for RA, but its underlying mechanism is unclear. In this study, we explore the effects of WJT on human RA-FLSs and the underlying molecular mechanism. Methods: The major components of WJT were determined using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Cell proliferative ability was evaluated by CCK-8, colony formation assay, and EdU incorporation assay. Cell apoptotic capacity was examined by caspase-3 and caspase-9 activity test. Protein levels of Bax and Bcl-2 were investigated by western blotting. High-throughput sequencing and bioinformatics analysis were conducted to screen and identify targeted genes, followed by identification by qRT-PCR and western blotting. Results: In this study, we have identified 346 compounds in WJT. Our results showed that WJT inhibited the RA-FLSs proliferation, and promoted apoptosis in a dose- and time-dependent manner. More importantly, 184 differentially expressed genes (DEGs) has been screened after WJT treatment based on DEGSeq2 and 278 DEGs was identified by DEGSeq2 combined with WGCNA. Then, 10 hub genes were identified based on two different analyses, while the expression levels of only SMC3 , THOC1 , BUB1 , and STAG2 were decreased after WJT treatment, which was identical to the sequencing profiles.Conclusions: WJT exerted its anti-proliferation and pro-apoptosis effects possibly through suppressing the expression of SMC3, THOC1, BUB1, and STAG2 in RA-FLSs. Thus, therapeutics targeting these genes may be a promising strategy for rescuing RA. Keywords: Rheumatoid arthritis; bioinformatics; fibroblast-like synoviocytes; Wantong Jingu Tablet

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