Bone marrow mesenchymal stem cell-derived exosomes alleviate skin fibrosis in systemic sclerosis by inhibiting the IL-33/ST2 axis via the delivery of microRNA-214.

Interleukin (IL)- 33 is a tissue-derive proinflammatory cytokine that promotes fibrosis in systemic sclerosis (SSc). microRNA (miR)- 214 expression has been elaborated to be downregulated in SSc patients and exert anti-fibrotic and anti-inflammatory effects. This study elucidates the role of bone marrow mesenchymal stem cell-derived exosome (BMSC-Exos)-delivered miR-214 in SSc and the relationship between this miR and IL-33/ST2 axis. SSc clinical samples were obtained to evaluate levels of miR-214, IL-33, and ST2. Primary fibroblasts and BMSC-Exos were extracted, followed by the co-culture of PKH6-labeled BMSC-Exos and fibroblasts. Subsequently, Exos extracted from miR-214 inhibitor-transfected BMSCs were co-cultured with TGF-ß1-stimulated fibroblasts, after which the expression of fibrotic markers, miR-214, IL-33, and ST2, as well as fibroblast proliferation and migration, was determined. A skin fibrosis mouse model was induced with bleomycin (BLM) and treated with BMSC-Exos. Collagen fiber accumulation, collagen content, α-SMA expression, and IL-33 and ST2 levels were examined in BLM-treated or IL-33-knockout mice. IL-33 and ST2 were upregulated and miR-214 was downregulated in SSc patients. Mechanistically, miR-214 targeted IL-33 and blocked the IL-33/ST2 axis. BMSC-Exos delivering miR-214 inhibitor augmented proliferation, migration, and fibrotic gene expression in TGF-ß1-stimulated fibroblasts. Similarly, IL-33 induced migration, proliferation, and fibrotic gene expression in fibroblasts via ST2. In BLM-treated mice, IL-33 knockout suppressed skin fibrosis, and BMSC-Exos delivered miR-214 to suppress the IL-33/ST2 axis, thus mitigating skin fibrosis. Conclusively, BMSC-Exos alleviate skin fibrosis through the blockade of the IL-33/ST2 axis by delivering miR-214.

Stattic alleviates pulmonary fibrosis in a mouse model of rheumatoid arthritis-relevant interstitial lung disease.

Approximately 20% of rheumatoid arthritis (RA) patients have RA-related interstitial lung disease (RA-ILD). Stattic, an STAT3 inhibitor, has been confirmed to be relevant to both RA and ILD. Therefore, this study explored the effect of Stattic on the progression of joint disease and pulmonary fibrosis in zymosan-treated female SKG mice, an established model for autoimmune arthritis. The experimental mice developed pulmonary interstitial pneumonia, which is similar to human cellular and fibrotic nonspecific interstitial pneumonia. Oral gavage of Stattic (60 mg/kg/d) was initiated 10 weeks after zymosan injection. Arthritis and lung fibrosis outcome scores decreased significantly following Stattic treatment. An obvious decrease in lung collagen levels, measured using hydroxyproline level determination and collagen staining, was detected after 6 weeks in Stattic-exposed mice with established disease. Stattic also dramatically restricted arthritis progression, based on joint evaluation. Transforming growth factor beta 1 (TGF-ß1) is a pivotal fibrosis-causing cytokine, used here to treat myofibroblasts, thereby establishing a lung fibrosis cell model. Stattic treatment can mitigate the TGF-ß1-triggered inflammatory response, myofibroblast activation, oxidative stress, and hyperproliferation by modulating the JAK1/STAT3 pathway. Our observations support a direct role of Stattic-inhibited STAT3 activation in lung fibrosis, which may be particularly relevant in the RA-ILD context.

Analysis of Clinical Efficacy of Clearing Heat and Dispelling Paralysis Soup in the Treatment of Osteoarthritis of the Knee Joint and Its Effect on Patients' Motor Function.

OBJECTIVE: To investigate the effects of clearing heat and dispelling paralysis soup for osteoarthritis of the knee joint on the motor function of the knee joint and the level of inflammation of the organism in patients. METHODS: One hundred and sixteen patients with knee osteoarthritis admitted from January 2020 to May 2021 were selected and randomly divided into 2 groups, 58 cases in the control group were treated with loxoprofen sodium dispersible tablets, and 58 cases in the experimental group were treated with Qinghe dispel paralysis soup on the basis of the control group and the patients' balance ability analysis, gait parameter change analysis, VAS, JOA, AIMS2-SF scale assessment, and serum index. The treatment effects of the two treatment regimens were analyzed by testing. RESULTS: The anterior-posterior axis, left-right axis, A2-A6, A4-A8, and circumferential axis of the experimental group were lower than those of the control group after treatment (P < 0.05); the step length of the experimental group was higher than that of the control group after treatment (P < 0.05), and there were no significant differences in step speed, double-support phase, and step width (P > 0.05), but both groups improved significantly compared with those before treatment (P < 0.05); the VAS score of the experimental group was lower than that of the control group after treatment. The VAS scores of the experimental group were lower than those of the control group, and the scores of JOA and AIMS2-SF were higher than those of the control group (P < 0.05); the levels of TIMP-1 in the experimental group were higher than those in the control group, and the levels of TNF-α, TLR4, MMP-3, and IL-1 were lower than those in the control group after treatment (P < 0.05); there was no significant difference in the incidence of adverse reactions between the two groups during treatment (P > 0.05), and the efficiency of the experimental group was higher than that of the control group (P < 0.05). CONCLUSION: Combined treatment with Qinghe dispel paralysis soup can better promote the recovery of balance, improve motor ability, and reduce the development of inflammation in the organism, with high safety and effectiveness.

miR-144-3p aggravated cartilage injury in rheumatoid arthritis by regulating BMP2/PI3K/Akt axis.

OBJECTIVES: Present study aimed to illustrate the role of miR-144-3p in rheumatoid arthritis (RA). METHODS: N1511 chondrocytes were stimulated by interleukin (IL)-1ß to mimic RA injury model in vitro. Rats were subjected to injection of type II collagen to establish an in vivo RA model, and the arthritis index score was calculated. Cell viability was determined by Cell Counting Kit-8. The expression of cartilage extracellular matrix proteins (collagen II and aggrecan) and matrix metalloproteinase protein were determined by quantitative real-time polymerase chain reaction and western blots. Cell apoptosis was measured by flow cytometry. Enzyme-linked immunosorbent assay was applied to test the secretion of pro-inflammatory cytokines (IL-1ß and tumour necrosis factor-α). Tissue injury and apoptosis were detected by haematoxylin-eosin staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling assay staining. Interaction of miR-144-3p and bone morphogenetic protein 2 (BMP2) was verified by dual-luciferase assay. RESULTS: miR-144-3p was dramatically increased in IL-1ß-induced N1511 cells. miR-144-3p depletion elevated cell viability, suppressed apoptosis, pro-inflammatory cytokine releasing, and extracellular matrix loss in IL-1ß-induced N1511 cells. Moreover, miR-144-3p targeted BMP2 to modulate its expression negatively. Activation of phosphatidylinositol 3-kinase (PI3K)/Akt signalling compromised inhibition of BMP2 induced aggravated N1511 cell injury with IL-1ß stimulation. Inhibition of miR-144-3p alleviated cartilage injury and inflammatory in RA rats. CONCLUSION: Collectively, miR-144-3p could aggravate chondrocyte injury inflammatory response in RA via BMP2/PI3K/Akt axis.

MiR-144-3p induced by SP1 promotes IL-1ß-induced pyroptosis in chondrocytes via PTEN/PINK1/Parkin axis.

Rheumatoid arthritis (RA) often leads to functional disabilities and deformities. MiRNA plays a vital role in cell pyroptosis. Nevertheless, the function and underlying mechanism of miR-144-3p in pyroptosis during the progression of RA remains unclear. In this study, N1511 cells were stimulated with IL-1ß to construct a RA model. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was performed to assess the cell viability. Cell pyroptosis was detected by flow cytometry. The levels of inflammatory cytokines (TNF-α, IL-6, and IL-18) were assessed by enzyme-linked immunosorbent assay (ELISA). The relationship among specific protein 1 (SP1), microRNA-144-3p (miR-144-3p), and phosphatase and tensin homolog (PTEN) was explored by dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP), respectively. The level of miR-144-3p in N1511 cells was upregulated by IL-1ß. MiR-144-3p knockdown inhibited IL-1ß-induced pyroptosis in N1511 cells, and the expressions of NOD-like receptor family pyrin domain containing 3 (NLRP3), Cleaved caspase-1, Gasdermin D (GSDMD), and Cleaved caspase-3 in IL-1ß-stimulated N1511 cells were increased. The levels of inflammatory cytokines in N1511 cells were increased by IL-1ß, which were restored by miR-144-3p knockdown. MiR-144-3p knockdown abolished IL-1ß-induced inactivation of putative kinase 1 (PINK1)/Parkin RBR E3 ubiquitin-protein (Parkin) signalling. Moreover, transcription factor SP1 could upregulate miR-144-3p expression and miR-144-3p negatively regulated PTEN expression. In summary, MiR-144-3p induced by SP1 could promote IL-1ß-induced chondrocyte pyroptosis via inhibiting PTEN expression and suppressing the activation of PINK1/Parkin signalling, which provided a new strategy against RA.

Correlation between NLR, PLR, and LMR and Disease Activity, Efficacy Assessment in Rheumatoid Arthritis.

OBJECTIVE: To analyze the value of neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and lymphocyte-monocyte ratio (LMR) in the evaluation of disease activity and efficacy in patients with rheumatoid arthritis (RA). METHODS: The clinical data of 132 newly diagnosed RA patients admitted to our hospital from November 2018 to January 2020 were retrospectively analyzed, and the NLR, PLR, and LMR were calculated. According to the 28-joint disease activity score (DAS28), all patients was divided into the remission group (n = 40) and the active group (n = 92). According to the curative effect of the active group, the patients were divided into the effective group (n = 61) and the ineffective group (n = 39). Logistic regression analysis of clinical data was to determine the influencing factors of RA disease activity. The receiver operating characteristic curve (ROC) was used to evaluate the predictive value of NLR, PLR, and LMR on disease activity and efficacy of RA. RESULTS: The number of cases of smoking history, the number of cases of drinking history, and NLR, PLR, CRP, and ESR levels of patients in the active group were higher than those of the remission group, and the LMR level was lower than that of the remission group; the differences were statistically significant (P < 0.05). The results of multivariate logistic regression analysis showed that NLR, PLR, LMR, CRP, and ESR were independent influencing factors of disease activity in RA patients (P < 0.05). The AUC of NLR, PLR, and LMR on the disease activity of RA patients was 0.872, 0.821, and 0.824, the sensitivity was 87.6%, 70.2%, and 69.3%, and the specificity was 75.6%, 76.8%, and 84.3%, respectively. The NLR and PLR values of the effective group were lower than those of the ineffective group, and the LMR values were higher than those of the ineffective group, and the differences were statistically significant (P < 0.05). The AUC of NLR, PLR, and LMR on the efficacy of RA patients was 0.756, 0.732, and 0.779, the sensitivity was 68.4%, 60.2%, and 67.9%, and the specificity was 83.2%, 86.4%, and 85.1%, respectively. CONCLUSION: NLR, PLR, and LMR are the independent factors that affect the disease activity of RA patients and can better evaluate the disease activity and efficacy of RA.

[Pirfenidone Inhibits Cytokines/chemokines Release from Alveolar Macrophages].

Objective To investigate the effect of pirfenidone on cytokine/chemokine production by alveolar macrophages(AMs)in patients with idiopathic nonspecific interstitial pneumonia(iNSIP)or idiopathic pulmonary fibrosis(IPF).Methods We prospectively enrolled 10 iNSIP patients,11 IPF patients,and 8 non-interstitial lung disease(non-ILD)patients(control group)from our center from January 2015 to December 2018.AMs from bronchoalveolar lavage fluid(BALF)were cultured with or without lipopolysaccharide(LPS)stimulation.The production of Th1 cytokines [soluble tumor necrosis factor receptor(sTNFR)-1,sTNFR-2,and interleukin(IL)-1ß],Th2 cytokines [IL-10 and granulocyte-macrophage colony-stimulating factor(GM-CSF)],angiogenic chemokines [IL-18 and macrophage inflammatory protein(MIP)-1ß],and angiostatic chemokines [interferon-gama inducible monokines(MIG)and interferon-gama inducible protein(IP-10)] in the culture supernatants were measured by a bead-based assay,Luminex.The effect of pirfenidone on the cytokine/chemokine production was tested at various concentrations(0,0.03,0.10,0.30 mg/ml).Results The spontaneous and LPS-stimulated release of TNF-α,sTNFR-1,sTNFR-2,IL-1ß,IL-10,MIP-1ß,MIG,and IP-10 by AMs were significantly increased in iNSIP and IPF groups compared with control group(all P<0.05),but no difference in IL-18 was seen among three groups(all P>0.05).MIG and IP-10 were significantly higher in iNSIP group than in IPF group(both P<0.05).Pirfenidone suppressed the spontaneous and LPS-stimulated AMs release of all studied cytokine/chemokine in iNSIP and IPF in a dose-dependent manner at concentrations of 0.10 and 0.30 mg/ml,and no difference was observed between iNSIP and IPF groups(both P>0.05).Conclusion Pirfenidone can markedly suppress cytokine/chemokine expression in iNSIP and IPF patients,but the difference is not significant between these two groups of patients.