Objective To establish an UPLC method for determination of Triptolide in serum and explore its pharmacokinetics in patients with rheumatoid arthritis after oral administration of tripterygium glycosides tablet. Methods Three patients with rheumatoid arthritis were enrolled. Using Estazolam as internal standard, serum was extracted with acetic ether, and determination was performed on column of Waters Acquity C18 (2.1 mm×100 mm, 1.7 μm) with mobile phase consisted of acetonitrile-0.1% glacial acetic acid (30∶70) at the flow rate of 0.2 mL/min. The column temperature was 30 ℃, and the detection wavelength was set at 220 nm. The serum concentration of Triptolide was processed by DAS 2.1.1 computer program. Results Triptolide was well-separated from internal standard, and the retention time were about 4.9 min and 8.9 min, respectively. Linear range of Triptolide was 13.13-840.00 ng/mL. RSD of intra-day and inter-day were lower than 15%and the recoveries were 88.25%-99.33%. Pharmacokinetic parameters were as follows:Cmax was (159.97±42.43) ng/mL, Tmax was (1.33±0.58) h, T1/2βwas (7.51±2.26) h, and AUC0-12 h was (1131.12±89.20) mg?h/L, respectively. Conclusion Pharmacokinetics of Triptolide conformed to two compartment model. Triptolide can be quickly absorbed, and exists differences among individuals.

Objective:To observe the effect of Xinfeng Capsule (XFC) on synovial membrane PI3K/AKT/mTOR pathway in adjuvant arthritis (AA) rats.Methods:Rats were randomly divided into normal control group,model control group,methotrexate group,tripterygium glycoside group and XFC group.The expressions of IL-6,IL-10,HIF-1α and VEGF-A were detected by immunohistochemical method,and the expressions of PI3K,AKT1 and p-AKT1 were detected by immunoblotting.Results:Expression of AKT1,mTOR and ES proteins in synovial blood vessels.The expression of IL-6,VEGF,HIF-1α and synovial membrane PI3K,AKT1,p-AKT1,mTOR,ES in the model group were significantly higher than those in the control group The expression of PI3K,p-AKT1,mTOR,ES in synovial membrane decreased,and IL-10 in serum increased in XFC group,and the expression of HIF-1α,IL-6 and VEGF-A were decreased.Conclusion:XFC can improve angiogenesis in AA synovium by regulating PI3K/AKT/mTOR signaling pathway,HIF-1α and ES expression.

Rheumatoid arthritis (RA), as a common systemic inflammatory autoimmune disease, affects approximately 1 in 100 individuals. Effective treatment for RA is not yet available because current research does not have a clear understanding of the etiology and pathogenesis of RA. Xinfeng Capsule, a patent Chinese herbal medicine, has been used in the treatment of RA in recent years. Despite its reported clinical efficacy, there are no large-sample, multicenter, randomized trials that support the use of Xinfeng Capsule for RA. Therefore, we designed a randomized, double-blind, multicenter, placebo-controlled trial to assess the efficacy and safety of Xinfeng Capsule in the treatment of RA.

Long non-coding RNAs(lncRNAs)can be involved in regulating gene expression through multiple interactions with DNA,RNA or proteins.Studies have shown that lncRNAs not only regulate innate immune response,but also regulate immune cell development and adaptive immunity.According to recent reports,a number of lncRNAs play a role in the pathogenesis of immune-mediated inflammatory diseases,including systemic lupus erythematosus(SLE),and may become a diagnostic marker and prognostic reference for diseases.This paper summarizes the research progress of lncRNAs in SLE,with the hope of providing more new ideas and methods for the study of SLE.

OBJECTIVE: To observe the changes of pulmonary function and Notch signaling pathway of lung tissues in adjuvant-induced arthritis rats, and to investigate the mechanism of reduced lung function. METHODS: A total of 30 rats were randomly divided into a normal group and a model group. Rats in the model group were induced to establish the adjuvant arthritis AA model by intradermally injecting 0.1 mL Freund's complete adjuvant into the right paw. After 30 days, we observed the paw edema volume, arthritis index, pulmonary function, histomorphology, and Notch receptor/ligand of the lung tissue. RESULTS: Compared with the normal group, the paw edema volume, arthritis index, average expiratory flow within 0.3 s (FEV0.3/FVC), and the level of Notch3, Notch4 and Jagged2 of the lung tissue in the model group was significantly increased, while maximum expiratory flow at 50% of vital capacity (FEF50), maximum expiratory flow at 75% of vital capacity (FEF75), forced expiratory flow (PEF) and the expression of Notch1 of Jagged1 and Delta1 in the lung were significantly decreased (P<0.05, P<0.01). There were significant positive correlations between FEV0.3/FVC and Notch4. FEV0.3/FVC, FEF25, FEF50 and Notch3, Delta1 were negatively correlated, respectively (P<0.05, P<0.01). CONCLUSION While arthritis occurs in AA rats, pulmonary function declines and significantly correlates with the expression of Notch receptor/ligand. The deposition of immune complex in the lung after the injection of CFA activates the Notch signaling pathway, and results in further decline of pulmonary function by signaling cascades.
Animals ; Arthritis, Experimental ; metabolism ; physiopathology ; Calcium-Binding Proteins ; metabolism ; Freund's Adjuvant ; Intercellular Signaling Peptides and Proteins ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Jagged-1 Protein ; Lung ; metabolism ; pathology ; Membrane Proteins ; metabolism ; Rats ; Receptors, Notch ; metabolism ; Respiratory Insufficiency ; metabolism ; physiopathology ; Serrate-Jagged Proteins ; Signal Transduction ; Vital Capacity

Objective To explore the effects of spleen-fortifying and kidney-replenishing therapy on man-ifestation and quality of life in patients with systemic lupus erythematosus disease ( SLE) . Methods 50 SLE patients were randomly divided into treatment group and control group. Both groups were given pred-nisone tablets combined with hydroxychloroquine sulfate while the treatment group also took the Spleen-fortifying and Kidney-replenishing Decoction ( Jianpi Yishen Tang ) . Changes of clinical manifestations and laboratory parameters before and after treatment were used to evaluate the efficacy and effects on the quality of life. Results Total effective rate of 88. 0% with 4 cases of clinical response, 10 cases of re-markable effect, 8 cases of effect and 3 cases of non-respond in the treatment group, was significantly higher than 72. 0% of the control group (clinical response 2 cases, remarkable effect 6 cases, effect 10 cases and non-response 7 cases) (P<0. 05). There was no significant difference between the two groups in SLEDAI scores. The treatment group was significantly superior to the control group (P<0. 01) in im-proving patients’ quality of life and scores of SDS and SAS. TCM symptom scores in the treatment group were significantly lower than those in the control group (P<0. 05). IgG, CRP and other laboratory pa-rameters in the treatment group were also significantly lower than that of the control group ( P<0 . 05 ) . Levels of IgM, C4, WBC, and RBC in the treatment group were significantly higher than those in the control group ( P<0 . 05 ) . Conclusion Spleen-fortifying and kidney-replenishing therapy could signifi-cantly attenuate clinical manifestations, disease activity, TCM symptom scores and relieve anxiety and depression in SLE patients. The efficiency and safety had been proved to improve the quality of patents’ life.

Objective To observe the mechanism of Jianpi (invigorating the spleen)Huashi (resovling dampness)Tongluo (removing obstruction in collaterals)Formula (JHTF)on PI3K/AKT/mTOR signa-ling pathway,hypoxia inducible factor (HIF-1α),vascular endothelial growth factor (VEGF-A),mi-crovessel density (MVD)and endostatin (ES)of synovial tissue in rats with adjuvant arthritis (AA). Methods Fifty rats were randomly divided into five groups — normal control group,model group, methotrexate group,tripterygium glycosides tablet group and JHTF group.Rat AA model was induced by using Freund's complete adjuvant.After modeling,each treatment group were i.g.corresponding medica-tion for consecutive 30 days,and then the expression of microvascular density (MVD)was detected using immunohistochemical,interleukin (IL)-6,IL-10,HIF-1αand VEGF were measured by enzyme-linked immunosorbent assay,and PI3K,AKT1,p-AKT1,mTOR and ES proteins were detected by Western blotting.Results Compared with normal control group,footpad swelling,arthritic index increased,ser-um IL-6,VEGF and HIF-1αand MVD,PI3K,AKT1,p-AKT1,mTOR,ES in synovial vascular signifi-cantly increased in the model group(P <0.05 or P <0.01).Compared with the model group,the MVD count,serum VEGF-A,HIF-1αand IL-6 decreased in JHTF group.The expressions of PI3K,p-AKT1, mTOR,ES in synovial vascular in JHTF group were lower ,and serum IL-10 was higher than those of the model control group.Conclusion Jianpi Huashi Tongluo Formula can improve angiogenesis by regula-ting PI3K/AKT/mTOR pathway,expression of HIF-1αand ES in rat AA model.

Objective To explore the therapeutic mechanism of Jianpi Zishen(JPZS)granules for systemic lupus erythematosus(SLE)in light of podocyte autophagy regulation.Methods TCMSP,GeneCards,OMIM,and TTD databases were used to obtain the targets of JPZS granules,SLE,and podocyte autophagy.The protein-protein interaction network was constructed using Cytoscape,and the key active ingredients and targets were screened for molecular docking.In the clinical study,46 patients with SLE were randomized into two groups to receive baseline treatment with prednisone acetate and mycophenolate mofetil(control group)and additional treatment with JPZS granules(observation group)for 12 weeks,with 10 healthy volunteers as the healthy control group.Urinary levels of nephrin and synaptopodin of the patients were detected with ELISA.Western blotting was performed to determine peripheral blood levels of p-JAK1/JAK1,p-STAT1/STAT1,LC3Ⅱ/LC3I,and p62 proteins of the participants.Results Four key active ingredients and 5 core target genes(STAT1,PIK3CG,MAPK1,PRKCA,and CJA1)were obtained,and enrichment analysis identified the potentially involved signaling pathways including AGE-RAGE,JAK/STAT,EGFR,and PI3K/Akt.Molecular docking analysis showed that STAT1 was the most promising target protein with the highest binding activity,suggesting its role as an important mediator for signal transduction after JPZS granule treatment.In the 43 SLE patients available for analysis,treatment with JPZS granule significantly reduced serum levels of p-JAK1/JAK1,p-STAT1/STAT1,and LC3Ⅱ/LC3I(P<0.05 or 0.01),increased the protein level of P62(P<0.05),and reduced urinary levels of nephrin and synaptopodin(P<0.05).Conclusion The therapeutic effect of JPZS granules on SLE is mediated probably by coordinated actions of quercetin,kaempferol,β-sitosterol,and isorhamnetin on their target gene STAT1 to inhibit the JAK/STAT pathway,thus suppressing autophagy and alleviating podocyte injuries in SLE.

Objective To explore the therapeutic mechanism of Jianpi Zishen(JPZS)granules for systemic lupus erythematosus(SLE)in light of podocyte autophagy regulation.Methods TCMSP,GeneCards,OMIM,and TTD databases were used to obtain the targets of JPZS granules,SLE,and podocyte autophagy.The protein-protein interaction network was constructed using Cytoscape,and the key active ingredients and targets were screened for molecular docking.In the clinical study,46 patients with SLE were randomized into two groups to receive baseline treatment with prednisone acetate and mycophenolate mofetil(control group)and additional treatment with JPZS granules(observation group)for 12 weeks,with 10 healthy volunteers as the healthy control group.Urinary levels of nephrin and synaptopodin of the patients were detected with ELISA.Western blotting was performed to determine peripheral blood levels of p-JAK1/JAK1,p-STAT1/STAT1,LC3Ⅱ/LC3I,and p62 proteins of the participants.Results Four key active ingredients and 5 core target genes(STAT1,PIK3CG,MAPK1,PRKCA,and CJA1)were obtained,and enrichment analysis identified the potentially involved signaling pathways including AGE-RAGE,JAK/STAT,EGFR,and PI3K/Akt.Molecular docking analysis showed that STAT1 was the most promising target protein with the highest binding activity,suggesting its role as an important mediator for signal transduction after JPZS granule treatment.In the 43 SLE patients available for analysis,treatment with JPZS granule significantly reduced serum levels of p-JAK1/JAK1,p-STAT1/STAT1,and LC3Ⅱ/LC3I(P<0.05 or 0.01),increased the protein level of P62(P<0.05),and reduced urinary levels of nephrin and synaptopodin(P<0.05).Conclusion The therapeutic effect of JPZS granules on SLE is mediated probably by coordinated actions of quercetin,kaempferol,β-sitosterol,and isorhamnetin on their target gene STAT1 to inhibit the JAK/STAT pathway,thus suppressing autophagy and alleviating podocyte injuries in SLE.

Rheumatoid arthritis(RA)is a common chronic autoimmune disease with synovitis as its pathological basis and erosive arthritis as its main symptom.Pathogenesis of RA is complex,combination of genetic factors,environmental factors,immune cells,cytokines and autoantibodies causes joint injury,bone destruction and multi-system disease of RA.However,the above mecha-nisms can not fully explain the poor prognosis,high disability rate and poor clinical treatment effect of RA.Therefore,exploring new pathogenesis and therapeutic targets of RA is the focus of RA research.In recent years,with the deepening of RA research,it has been found that there is a new form of cell death in pathological process of RA,namely ferroptosis.Ferroptosis is a type of cell death caused by inhibition of glutathione peroxidase activity and accumulation of lipid reactive oxygen species.Previous studies have con-firmed the close correlation between RA and ferroptosis,this paper mainly explores ferroptosis-related signal pathways that affect the change and development of RA disease from the perspective of regulating the main signal pathways of ferroptosis,so as to find new therapeutic targets for RA and new therapeutic ideas for research.