Abstract Background The dysfunction of osteoblasts has been identified as a pivotal contributor to bone erosion in rheumatoid arthritis (RA). Emerging evidence shows long non-coding RNAs (lncRNAs) as important regulators in various pathological processes, including RA. However, the specific regulatory mechanisms of lncRNAs in osteoblast dysfunction of RA remain poorly understood and worth further investigation. Methods Collagen induced arthritis (CIA) rat model was established, tarsal tissue was isolated for sequencing of lncRNAs. The differentially expressed lncRNA was screened and verified in vivo and in vitro. The target microRNA of lncRNA and the target protein of microRNA were predicted by bioinformatics analysis and validated. The biology function of the lncRNA and microRNA was further investigated by knockdown or overexpression experiment in murine osteoblastic cell MC3T3-E1. Results LncRNA MIAT was highly expressed in tarsal tissue and serum of CIA rat as well as TNF-α-induced osteoblasts, and its high expression was negatively correlated with the expression of bone formation marker osteocalcin. Functionally, knockdown of lncRNA MIAT promoted the proliferation and mineralization of osteoblasts. Mechanistically, lncRNA MIAT directly bound to let-7i-5p and exerted a negative regulatory influence on its expression. CKIP-1 was a target of let-7i-5p. Overexpression of let-7i-5p promoted osteoblast proliferation and mineralization by targeting CKIP-1. Conclusions Taken together, this study demonstrated that lncRNA MIAT was involved in the regulation of osteoblast proliferation and mineralization through the lncRNA MIAT/let-7i-5p/CKIP-1 axis in RA.

IntroductionPathological changes in the articular cartilage (AC) and synovium are major manifestations of osteoarthritis (OA) and are strongly associated with pain and functional limitations. Exosome-derived microRNAs (miRNAs) are crucial regulatory factors in intercellular communication and can influence the progression of OA by participating in the degradation of chondrocytes and the phenotypic transformation in the polarization of synovial macrophages. However, the specific relationships and pathways of action of exosomal miRNAs in the pathological progression of OA in both cartilage and synovium remain unclear.MethodsThis study evaluates the effects of fibroblast-like synoviocyte (FLS)-derived exosomes (FLS-Exos), influenced by miR-146a, on AC degradation and synovial macrophage polarization. We investigated the targeted relationship between miR-146a and TRAF6, both in vivo and in vitro, along with the involvement of the NF-κB signaling pathway.ResultsThe expression of miR-146a in the synovial exosomes of OA rats was significantly higher than in healthy rats. In vitro, the upregulation of miR-146a reduced chondrocyte apoptosis, whereas its downregulation had the opposite effect. In vivo, exosomes derived from miR-146a-overexpressing FLSs (miR-146a-FLS-Exos) reduced AC injury and chondrocyte apoptosis in OA. Furthermore, synovial proliferation was reduced, and the polarization of synovial macrophages shifted from M1 to M2. Mechanistically, the expression of TRAF6 was inhibited by targeting miR-146a, thereby modulating the Toll-like receptor 4/TRAF6/NF-κB pathway in the innate immune response.DiscussionThese findings suggest that miR-146a, mediated through FLS-Exos, may alleviate OA progression by modulating cartilage degradation and macrophage polarization, implicating the NF-κB pathway in the innate immune response. These insights highlight the therapeutic potential of miR-146a as a protective agent in OA, underscoring the importance of exosomal miRNAs in the pathogenesis and potential treatment of the disease.

Glucocorticoid-induced osteonecrosis of the femoral head (GIONFH) is deeply relevant to damage and dysfunction of bone microvascular endothelial cells (BMECs). Recently, necroptosis, a newly programmed cell death with necrotic appearance, has garnered increasing attention. Luteolin, a flavonoid compound derived from Rhizoma Drynariae, has numerous pharmacological properties. However, the effect of Luteolin on BMECs in GIONFH through the necroptosis pathway has not been extensively investigated. Based on network pharmacology analysis, 23 genes were identified as potential targets for the therapeutic effect of Luteolin in GIONFH via the necroptosis pathway, with RIPK1, RIPK3, and MLKL being the hub genes. Immunofluorescence staining results revealed high expression of vWF and CD31 in BMECs. In vitro experiments showed that incubation with dexamethasone led to reduced proliferation, migration, angiogenesis ability, and increased necroptosis of BMECs. However, pretreatment with Luteolin attenuated this effect. Based on molecular docking analysis, Luteolin exhibited strong binding affinity with MLKL, RIPK1, and RIPK3. Western blotting was utilized to detect the expression of p-MLKL, MLKL, p-RIPK3, RIPK3, p-RIPK1, and RIPK1. Intervention with dexamethasone resulted in a significant increase in the p-RIPK1/RIPK1 ratio, but the effects of dexamethasone were effectively counteracted by Luteolin. Similar findings were observed for the p-RIPK3/RIPK3 ratio and the p-MLKL/MLKL ratio, as anticipated. Therefore, this study demonstrates that Luteolin can reduce dexamethasone-induced necroptosis in BMECs via the RIPK1/RIPK3/MLKL pathway. These findings provide new insights into the mechanisms underlying the therapeutic effects of Luteolin in GIONFH treatment. Additionally, inhibiting necroptosis could be a promising novel approach for GIONFH therapy.

Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease, and the mechanism of SLE is yet to be fully elucidated. The aim of this study was to explore the role of two-pore segment channel 2 (TPCN2) in SLE pathogenesis. Methods: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of TPCN2 in SLE. We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell. Knockdown of TPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting. Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation, apoptosis, and cell cycle of TPCN2-deficient cells. In addition, gene expression profile of TPCN2-deficient cells was analyzed by RNA sequencing (RNA-seq). Results: TPCN2 knockdown with short hairpin RNA (shRNA)-mediated lentiviruses inhibited cell proliferation, and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells. We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells, and screened the differential genes, which were enriched for the G2/M checkpoint, complement, and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways, as well as changes in levels of forkhead box O, phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin, and T cell receptor pathways; moreover, TPCN2 significantly influenced cellular processes and biological regulation. Conclusion: TPCN2 might be a potential protective factor against SLE.

The liver is essential for metabolic and immune functions and has been linked to systemic inflammatory diseases. However, the role of the liver is still elusive during the development of rheumatoid arthritis (RA), although there have been indeed some reports. We used label-free quantitative proteomics and experimental verification in this study to reveal the hepatic lipid metabolism and immune function during collagen-induced arthritis (CIA) development. The proteomics results revealed that the role of the liver differs in different phases of CIA rats. In terms of specific performance, hepatic lipid metabolism, which is primarily concerned with cholesterol, triacylglycerol, and phospholipid, was significantly influenced in the CIA induction phase, whereas the immune function, which includes binding of granulocytes, adhesion of immune cells, etc., was affected considerably at the peak phase of CIA rats compared to normal rats. Finally, the hepatic dynamic changes in CIA rats were further confirmed using targeted metabolomics and ELISA. We found that most fatty acids of the liver in the CIA induction phase were significantly decreased, and proteins related to complement activation and migration or adhesion of immune cells including C3, MMP-8, CTSZ, and S100A9 were significantly increased in the liver of CIA rats in the peak phase. Our findings indicated that the lipid metabolism and immune function of the liver were influenced in CIA rats. Thus, the conditions of the liver during RA development should be considered in therapeutic and nutritional interventions.

目的:探讨舒筋活络方(SHC)对Toll样受体(TLRs)介导的滑膜炎症信号转导通路及其下游炎症因子的调节作用及其相关性.方法:将高、中、低浓度的SHC分别加入脂多糖(LPS)诱导的滑膜成纤维细胞(FLS),采用流式细胞术(FCM)对固有免疫代表性元件TLR2、TLR4的表达进行测定,一氧化氮(NO)一步法、酶联免疫吸附测定(ELISA)法对下游炎症因子NO和白介素6(IL-6)浓度进行测定,采用SPSS 20.0软件进行相关性分析.结果:与空白组比较,模型组TLR2和TLR4表达显著增强(P＜0.05),NO、IL-6释放量显著升高(P＜0.05);与模型组比较,SHC各浓度组均不同程度抑制TLR2、TLR4表达及NO、IL-6释放(P＜0.05),TLR2、TLR4阳性比例及NO、IL-6释放量随SHC浓度升高而降低;TLR2、TLR4和NO、IL-6之间均呈显著的正相关性(P＜0.05).结论:SHC通过调控固有免疫反应关联网络中互相紧密联系的各元件来调节骨关节炎(OA)进程,为中药复方影响OA病理机制的阐明提供了新的数据.

Tendinopathy refers to a type of tendon disease with a multifactorial spectrum. Recent research has begun to reveal the effects of inflammation on the tendinopathic process, especially in the first stage of tendinopathy. Radial extracorporeal shock wave therapy (rESWT) has been successfully used to treat orthopedic diseases. However, the molecular mechanisms underlying the anti-inflammatory effects of rESWT on tumor necrosis factor-α treated tenocytes have not been fully elucidated. In this study, we applied total protein tandem mass tag-labeled quantitative proteomics with liquid chromatography-mass spectrometer/mass spectrometer technology to identify differentially expressed proteins (DEPs) among inflammatory tenocytes, rESWT inflammatory tenocytes, and controls using three biological replicates. Human tenocytes were used and they were cultured in vitro. In total, 1028 and 40 DEPs were detected for control versus inflammatory tenocytes and for inflammatory tenocytes versus rESWT inflammatory tenocytes, respectively. Further, we identified integrin α2, selenoprotein S, and NLR family CARD domain-containing protein 4 as pivotal molecular targets of the anti-inflammatory effects of rESWT. This is the first study to provide a reference proteomic map for inflammatory tenocytes and rESWT inflammatory tenocytes. Our findings provide crucial insight into the molecular mechanisms underscoring the anti-inflammatory effects of rESWT in tendinopathy.

Background: The disruption of joint homeostasis is a critical event during the process of joint injury in osteoarthritis (OA). As regulatory molecules, microRNAs (miRNAs) can be released from secretory cells and delivered to recipient cells through extracellular vesicles (EVs), thereby playing an important role in regulating joint homeostasis. We hypothesized that the fibroblast-like synoviocytes (FLSs) in healthy joints could release EVs enriched in miRNAs that can maintain joint homeostasis by regulating the signal transduction pathways in the joints, whereby the articular cartilage (AC) is protected from degeneration, and OA progression is delayed. Methods: Via high-throughput sequencing and qPCR, we found that miR-150-3p was enriched in the circulating EVs in healthy rats. Next, we established an in vitro cell model in which chondrocytes were cultured with (i) FLSs transfected with miR-150-3p mimics or (ii) EVs released by FLSs (FLS–EVs) inside the healthy synovial membrane (SM). The transportation mechanism from FLSs to chondrocytes was studied using the EV inhibitor GW4869, and the FLSs were transfected with a miR-150-3p mimic or inhibitor. To assess the therapeutic effect of miR-150-3p-carrying EVs (EVs-150) in vivo, healthy FLS-derived EVs (H-FLS–EVs) were injected into the tail vein of rats with OA at various stages of the pathogenesis and evaluated for the progression of OA. Results: The chondrocytes could uptake fluorescent-labeled miR-150-3p mimics and FLS–EVs, and GW4869 suppressed this uptake. The overexpression of miR-150-3p could significantly reduce the concentrations of pro-inflammatory cytokines in the cell culture medium and the expression of the miR-150-3p target T cell receptor-interacting molecule 14 (Trim14), as well as the innate immune-related factors, including nuclear factor kappa B (NF-κB) and interferon-β (IFN-β). Similarly to the in vitro findings, the miR-150-3p level in the serum EVs was significantly upregulated among the EV-treated rats. In the AC of the OA rat model injected with H-FLS–EVs, the joint degeneration was suppressed, and Type II collagen (COLII) and aggrecan (ACAN) were significantly upregulated, whereas the innate immune-related factors Trim14, NF-κB, and IFN-β were downregulated compared with the levels in the untreated OA rats. Notably, the suppression of joint degeneration was more significant when H-FLS–EVs were administered at the early stages of OA rather than the late stages. Conclusion: H-FLS–EVs protect chondrocyte function and maintain joint homeostasis by modulating the innate immune response by suppressing the Trim14/NF-κB/IFNβ axis. These effects are achieved through the EV-mediated transport of miR-150-3p from the FLSs to the chondrocytes. Our findings show that EV-mediated miR-150-3p can be used to suppress OA, thus providing a novel therapeutic strategy. Additionally, the EV-mediated miR-150-3p transport may also serve as a potential biomarker in the diagnosis, treatment, and prognosis of OA.

Background and objective: Omalizumab is a bio-targeted agent approved as add-on therapy for the treatment of severe asthma. Most patients with severe asthma show no response to omalizumab. American Thoracic Society (ATS) and European Respiratory Society (ERS) recommend blood eosinophil count and fractional exhaled nitric oxide (FeNO) as biomarkers with high value for increased response to omalizumab and periostin as a biomarker with a low value. In this study, we aimed to identify the biomarkers for predicting treatment response to omalizumab by performing whole blood transcriptional expression profiling using array and clinical data from GSE134544. Methods: We analyzed GSE134544 whole blood transcriptional and clinical data of omalizumab treatment using xCell, weighted gene co-expression network analysis (WGCNA), gene ontology enrichment analysis, KEGG pathway analysis, protein-protein interaction (PPI) network, and logistic regression analysis. Results: We calculated the immune enrichment score using xCell and found that CD4(+) T cells, CD4(+) Tem, CD4(+) memory T cells, CD8(+) Tcm, and dendritic cells (DC) were relatively higher in responders than in non-responders. Analysis of omalizumab response using WGCNA revealed that the above-mentioned significant immune cells in the red module was relevant to the sample traits; there were 547 genes in the red module. We identified 20 hub genes for the PPI network using cytoHubba, a Cytoscape plugin. Using logistic regression analysis, CD3E was found to be the only significant biomarker, and the area under the curve of ROC curves was 0.763. Conclusion: CD3E maybe a new predictive biomarker of response to omalizumab treatment in asthma patients and be used to select more suitable asthma patients for omalizumab treatment.

Background Reduced exercise tolerance is an important clinical feature of chronic obstructive pulmonary disease (COPD) and is associated with poor prognosis. The 6-min walk test (6MWT) is widely used to assess exercise capacity; however, it is not commonly administered in primary medical institutions because it requires a suitable site and professional training. Ultrasound has great potential for evaluating skeletal muscle dimensions in COPD. However, whether skeletal muscle ultrasound can predict impaired exercise tolerance is unclear. Methods The study included 154 stable patients with COPD, who were randomly divided into a development set and a validation set. The thickness (RF thick ) and cross-sectional area (RF csa ) of the rectus femoris were measured using ultrasound. Standardized RF thick (STD- RF thick ) and Standardized RF csa (STD-RFcsa) were obtained via standardization of RF thick and RF csa by patients' height. Results Strong correlations were observed between the 6MWD and RF thick (r = 0.84, p < 0.001) and between the 6MWD and RF csa (r = 0.81, p < 0.001). In the development set, the optimal cut-off values for men and women for predicting poor exercise tolerance were < 3.098 cm/m and < 3.319 cm/m for STD-RF thick and < 4.052 cm 2 /m and < 4.366 cm 2 /m for STD-RF csa , respectively. In the validation set, the area under the curve (AUC) values for the prediction of a 6MWD < 350 by STD-RF thick and STD-RF csa were 0.881 and 0.903, respectively. Finally, the predictive efficacy of STD-RF thick (AUC: 0.922), STD-RF csa (AUC: 0.904), and the derived nomogram model (AUC: 0.98) for exercise tolerance was superior to that of the sit-to-stand test and traditional clinical features. Conclusions Rectus femoris ultrasound has potential clinical application to predict impaired exercise tolerance in patients with COPD.

Asthma is driven by group 2 innate lymphoid cells, antigen-specific CD4+ T helper type 2 cells and their cytokines such as interleukin (IL)-4, IL-5, IL-13. IL-37 is decreased in asthma and negatively related to Th2 cytokines and other pro-inflammatory cytokines. Our study showed that IL-37 level in asthmatic peripheral blood mononuclear cells was lower than in healthy. Further, IL-37 was negatively correlated with exhaled nitric oxide, asthma control test score, atopy and rhinitis history in asthmatics. Then an OVA-induced asthma mice model treated with rhIL-37 was established. An antibody array was employed to uncover altered cytokines induced by IL-37 in mice lung tissue. 20 proteins differentially expressed after rhIL-37 treatment and five of them were validated in asthmatic peripheral blood mononuclear cells. Consistent with cytokine antibody array, CCL3, CCL4, CCL5 decreased after IL-37 administration. While CXCL9 and CXCL13 were no change. We concluded that IL-37 reduce asthmatic symptoms by inhibit pro-inflammatory cytokine such as CCL3, CCL4, CCL5.

目的:测定舒筋活络方(SHC)对骨关节炎(OA)患者滑膜成纤维细胞(FLS)增殖、迁移、侵袭的影响,明确其作用机制.方法:对胶原酶法分离培养的FLS进行形态学观察,通过MTT比色法、划痕实验、Transwell实验、ELISA法等技术测定SHC对FLS增殖、迁移、侵袭等的影响.结果:体外分离的FLS生长状态良好,具备成纤维细胞形态特性;所选药物浓度范围对FLS增殖影响较小,与空白组比较,较高浓度和低浓度的SHC对FLS增殖的影响差异均有统计学意义(P<0.05),FLS的增殖与SHC呈浓度依赖性;镜下观察到OA-FLS的迁移现象,其中,模型组48 h、72 h的迁移率与0 h时比较,差异有统计学意义(P<0.05),SHC组72 h的迁移率与0 h时比较差异有统计学意义(P<0.05),与模型组比较,48 h、72 h时,SHC组FLS迁移能力比较,差异有统计学意义(P<0.05);SHC组FLS侵袭穿过小室膜的细胞个数较空白组明显减少(P<0.05);SHC各浓度组均对MMP3的释放有不同程度的抑制作用(P<0.05).结论:SHC调节FLS增殖、抑制FLS的迁移和侵袭,维护和修复OA患者关节内组织完整性,为OA的关节保护提供新的预防和治疗策略.

目的:研究差异浓度的舒筋活络方(SHC)对骨关节炎(OA)滑膜成纤维细胞(FLS)的活性、凋亡及细胞周期的影响,进一步明确SHC治疗OA的作用机制.方法:以差异浓度的SHC干预胶原酶法消化分离培养的FLS,通过细胞增殖-毒性检测试剂盒(CCK-8)和流式细胞术(FCM)分别检测FLS的活性、凋亡和细胞周期.结果:与对照组比较,高、低浓度的SCH均不同程度的抑制FLS活性(P＜0.05),呈浓度依赖性,中浓度的SCH则对FLS活性有部分促进作用;SCH的各浓度组均不同程度的诱导FLS早期凋亡与晚期凋亡(P＜0.05),提高总凋亡率(P＜0.05);并且延长FLS的G0/G1期(P＜0.05),同时缩短G2/M期和S期(P＜0.05),呈明显的浓度依赖性,其中,高浓度组对FLS凋亡的诱导及周期的阻滞作用最明显.结论:差异浓度的SHC通过对FLS活性、凋亡和周期的调控抑制FLS的浸润性生长,减缓OA滑膜炎症反应.

Abstract Background Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovitis. Pien Tze Huang (PZH) is a Chinese patent medicine with anti-inflammatory and immunomodulatory effects. However, whether PZH could be used in RA therapy is still unknown. Therefore, this study aimed to explore the therapeutic effect and the potential mechanism of PZH on collagen-induced arthritis (CIA) mice. Methods Male DBA/1J mice were used to establish an animal model of CIA and then treated with different doses of PZH for 4 weeks. The therapeutic effect of PZH on CIA mice was evaluated by arthritis score, pathological staining, and detecting the levels of inflammatory factors in serum and joints. To investigate its possible mechanism, the activity of NF-κB signaling pathway, NLRP3 inflammasome and the level of A20 were detected. Results The results showed that PZH could alleviate the erythema and swelling of hind paws of CIA mice, improve the pathological conditions of joint and decrease the production of IL-1β, IL-6 and IL-17 in serum and joints. Furthermore, PZH could significantly inhibit the activity of NF-κB signaling pathway and NLRP3 inflammasome in the ankle joint of CIA mice compared with the model group. It also increased the level of A20 in the ankle joint of CIA mice. Conclusion This study indicated that PZH could alleviate the joint inflammation of CIA mice, and the mechanism might be related to the regulation of NF-κB signaling pathway and NLRP3 inflammasome.

Rheumatoid arthritis (RA) is a chronic disease with complex molecular network of pathophysiology, single drug is usually not full satisfactory because it is almost impossible to target the whole molecular network of the disease. Drug combinations that act synergistically with each another is an effective strategy in RA therapy. In this study, we aimed to establish a new strategy to search effective synergized compounds from Chinese herbal medicine (CHM) used in RA. Based on multi-information integrative approaches, imperatorin (IMP) and β-sitosterol (STO) were predicted as the most effective pair for RA therapy. Further animal experiments demonstrated that IMP+STO treatment ameliorated arthritis severity of collagen-induced arthritis (CIA) rats in a synergistic manner, whereas IMP or STO administration separately had no such effect. RNA sequencing and IPA analysis revealed that the synergistic mechanism of IMP+STO treatment was related to its regulatory effect on 5 canonical signaling pathways, which were not found when IMP or STO used alone. Moreover, LTA, CD83, and SREBF1 were 3 important targets for synergistic mechanism of IMP+STO treatment. The levels of these 3 genes were significantly up-regulated in IMP+STO group compared to model group, whereas IMP or STO administration separately had no effect on them. In conclusion, this study found that IMP and STO were 2 synergistic compounds from the CHM in RA therapy, whose synergistic mechanism was closely related to regulate the levels of LTA, CD83, and SREBF1.

Background: Asthma has become the most common chronic disease in children, which seriously affects children's health and growth. Yu-Ping-Feng powder (YPFP) is widely used for the treatment of asthma in children, but there are few meta-analyses to assess the add-on effects of YPFP in children with asthma. Therefore, it is necessary to conduct a systematic review to evaluate the efficacy and safety of YPFP in the management of asthma in children. Methods: PubMed, Cochrane Central Register of Controlled Trials (CENTRAL), EMBASE, Web of Science and the Chinese electronic databases including China Network Knowledge Infrastructure (CNKI), Chinese Biomedicine (CBM), Chinese Scientific Journals Database (VIP), and Wan Fang Database were searched for the randomized controlled trials (RCTs) of YPFP in children with asthma based on the eligibility criteria from the date of the database inception to 28 November 2018. Two reviewers assessed the articles and extracted data from the included RCTs independently. Data will be synthesized by either the fixed-effects or random-effects model according to a heterogeneity test. We will assess the risk of bias with the Cochrane Collaboration Tool and overall quality of the evidence using the Grading of Recommendations Assessment, Development and Evaluation system (GRADE). Primary outcomes include the improvement of symptoms including breathlessness, coughing, wheezing and the frequency of asthma exacerbations. Lung function, serum IgE level, blood eosinophil count, phlegm eosinophil count and adverse events will be assessed as the secondary outcomes. We will perform the data synthesis, sensitivity analyses, and subgroup analyses in the Rev-Man version 5.3 software. A funnel plot will be established to evaluate reporting bias. Results: This systematic review and meta-analysis will review and synthesis current clinical evidence of YPFP for the treatment of asthma in children. Conclusion: This analysis will provide high quality evidence of YPFP for the treatment of asthma in children. PROSPERO registration number: CRD42018111223.

The Chinese formula Pien Tze Huang (PZH) has been used to treat hepatocellular carcinoma (HCC) and showed positive clinical effects. However, the antitumor mechanism of PZH in HCC remains unclear. In this study, HCC xenograft Balb/c mice were treated with PZH; then, proteomics detection and Ingenuity Pathway Analysis (IPA) were used to analyze the differentiated phosphorylated proteins in tumor tissues. The results indicated that PZH could inhibit tumor weight by 50.76%. Eighty-four upregulated and 11 downregulated phosphorylated proteins were identified in PZH-treated mice. Twenty signaling pathways were associated with inflammation (including the IL-6 and TNFR1/2 pathways), cancer growth (including the p53 and FAK pathways), and the cell cycle (including the G2/M and G1/S checkpoint regulation pathways). Moreover, TNF-α, IL-6, and several typical differentially expressed phosphorylated proteins (such as p-CCNB1, p-FOXO3, and p-STAT3) in tumor tissues, tumor cell viability, and cell cycle arrest assay in vitro further verify the results of IPA. These results revealed that PZH achieved antitumor activity in HCC; the underlying mechanisms of which were mainly through regulating the inflammation-associated cytokine secretion, cancer growth pathways, and induction of G2/M arrest. These data provided the potential molecular basis for PZH to act as a therapeutic drug or a supplement to chemotherapy drugs for human HCC in the future.

BACKGROUND:Asthma is characterized to chronic airway inflammation. However, the role of oxidative damage and DNA damage in the pathophysiology of asthma have rarely been studied. On the other hand, there are evidences that DNA-dependent protein kinase (DNA-PK) participates in DNA damage repair and regulates innate immune responses and proinflammatory signaling pathways.METHODS:After ovalbumin (OVA)-induced asthmatic murine model was established, airway hyper-responsiveness (AHR), total and differential bronchoalveolar lavage fluid (BALF) cell counts. IL-4, IL-8, IL-13 and TNF-α were chosen to evaluate the airway inflammation, and oxidative damage indicators levels (8-isoprostane and 8-OhdG) in BALF were measured. Alkaline comet assay was conducted to detected DNA damage. Histological analysis was conducted after hematoxylin and eosin (HE) straining, and proteins were extracted for 3-nitrotyrosine (3-NT) detection and immunoblotting.RESULTS:AHR, infiltration of inflammatory cells and pro-inflammatory cytokine levels in lungs were significantly higher in asthmatic mice. OVA challenge resulted in robust increase in 3-NT, 8-isoprostane and 8OHdG in lungs, which represented oxidative damage level. DNA damage and repair proteins levels in asthma were also increased. NU7441 aggravated the DNA damage level. However, it suppressed infiltration of lung inflammatory cells and inflammatory cytokine levels, suggesting that DNA-PK may be a potential target for treatment of allergic asthma.CONCLUSIONS:Our study showed that oxidative damage and DNA damage existed in the airway of asthmatic mice. NU7441 augmented DNA damage level, and moreover, it also attenuated infiltration of inflammatory cells and pro-inflammatory cytokine levels in asthmatic lungs.

Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system (CNS). Currently, there is still lack of curative treatment for MS. Mesenchymal stem cell- (MSC-) based therapy is recently the subject of intense interest in autoimmune diseases. Here, we investigated the therapeutic effect and potential mechanism of human amnion mesenchymal cells (hAMC) on inflammation and remyelination in experimental autoimmune encephalomyelitis (EAE) mice. C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein (MOG) 35–55 peptide. hAMC were injected intraperitoneal when EAE was successfully established. The results demonstrated that application of hAMC significantly ameliorated the disease severity and histopathological changes in EAE mice. The production of proinflammatory cytokines such as IFN-γ, TNF-α, IL-1β, and IL-17A in the spleen and CNS was dramatically inhibited. Moreover, CD4+ T cells and CD8+ T cells in the CNS were also significantly decreased in EAE mice after hAMC treatment. In addition, hAMC treatment also promoted the production of neuron-repair factors (NGF, CNTF, and BDNF) in the CNS of EAE mice. In conclusion, these results indicated that hAMC could attenuate the inflammation and promote the remyelination in EAE mice, which might be a promising cell source for the therapy of MS.

Yupingfeng San (YPFS) is a representative Traditional Chinese Medicine (TCM) formula with accepted therapeutic effect on Asthma. However, its action mechanism is still obscure. In this study, we used network pharmacology to explore potential mechanism of YPFS on asthma. Nucleotide-binding oligomerization domain (NOD)-like receptor pathway was shown to be the top one shared signaling pathway associated with both YPFS and asthma. In addition, NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome was treated as target protein in the process of YPFS regulating asthma. Further, experimental validation was done by using LPS-stimulated U937 cells and ovalbumin (OVA)-sensitized BALB/c mice model. In vitro experiments showed that YPFS significantly decreased the production of TNF-α and IL-6, as well as both mRNA and protein levels of IL-1β, NLRP3, Caspase-1 and ASC in LPS-stimulated U937 cells. In vivo experiment indicated that YPFS treatment not only attenuated the clinical symptoms, but also reduced inflammatory cell infiltration, mucus secretion and MUC5AC production in lung tissue of asthmatic mice. Moreover, YPFS treatment remarkably decreased the mRNA and protein levels of IL-1β, NLRP3, Caspase-1 and ASC in lung tissue of asthmatic mice. In conclusion, these results demonstrated that YPFS could inhibit NLRP3 inflammasome components to attenuate the inflammatory response in asthma.