1.Identification and Biological Characterization of Pathogen and Screening of Effective Fungicides for Wilt of Tetradium ruticarpum
Yuxin LIU ; Qin XU ; Yue YUAN ; Tiantian GUO ; Zheng'en XIAO ; Shaotian ZHANG ; Ming LIU ; Fuqiang YIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):198-206
ObjectiveTo identify the pathogen species responsible for the wilt disease of Tetradium ruticarpum in Chongqing, investigate there biological characteristics, and screen effective fungicides, so as to provide a theoretical basis for disease control in production. MethodsThe pathogen was isolated via the tissue culture method. Pathogenicity was verified according to Koch's postulates. The pathogen was identified based on morphological characteristics and multi-gene phylogenetic analysis. The mycelial growth rate method was used for biological characterization of the pathogen and fungicide screening. ResultsThe pathogen colonies were nearly circular with irregular edges, white, short, velvety aerial hyphae, and pale purple undersides. Macroconidia were colorless, sickle-shaped, with 3-5 septa, while microconidia were transparent, elliptical, aseptate or with 1-2 septa. Multi-gene phylogenetic analysis showed that the pathogen clustered in the same clade as Fusarium fujikuroi with 100% support, which, combined with morphological characteristics, identified the pathogen causing wilt of T. ruticarpum in Chongqing as F. fujikuroi. The optimal conditions for the mycelial growth of F. fujikuroi were mung bean agar (MBA) with glucose as the carbon source, beef extract and yeast powder as nitrogen sources, 28 ℃, pH 7.0, and alternating light/dark conditions. The optimal conditions for sporulation were potato dextrose agar (PDA) with glucose as the carbon source, beef extract as the nitrogen source, 28 ℃, pH 7.0, and complete darkness. Among chemical fungicides, phenazine-1-carboxylic acid exhibited the strongest inhibitory effect on F. fujikuroi. Shenqinmycin and tetramycin were the most effective bio-fungicides. ConclusionThis study is the first to report F. fujikuroi as the causal agent of wilt disease in T. rutaecarpa. The chemical fungicide phenazine-1-carboxylic acid and the bio-fungicides shenqinmycin and tetramycin showed strong inhibitory effects against F. fujikuroi.
2.Analysis of regulation of prognosis,immune infiltration,and ferroptosis in sarcoma based on stemness index model
Jingxian WEI ; Lian MENG ; Hao SUN ; Tiantian ZHANG ; Chunxia LIU
Chinese Journal of Tissue Engineering Research 2025;29(19):4151-4160
BACKGROUND:The stemness index may be associated with the prognosis and immune infiltration of sarcoma,but the specific regulatory mechanism and characteristic genes have yet to be fully elucidated. OBJECTIVE:To investigate the correlation between stem cells and prognosis as well as immune infiltration in sarcoma employing the gene stemness index model and to identify the ferroptosis signature genes associated with sarcoma stem cells. METHODS:The sarcoma RNA sequencing data and related clinical information were obtained from the Cancer Genome Atlas(TCGA).The sarcoma RNA sequencing data were grouped using the sarcoma stemness index.Survival data were used to analyze prognosis between groups.Differentially expressed genes were obtained for pathway enrichment and immune infiltration analysis.Ferroptosis-related differential genes were used to construct a protein interaction network and analyze prognostic correlation.Rhabdomyosarcoma cell lines were cultured and divided into adherent cell group and stem cell group.The adherent cell group received no intervention,while the stem cell group was treated with serum-free culture to enrich stem cells in rhabdomyosarcoma cells.qRT-PCR was used to evaluate stemness markers,ferroptosis-related genes,and mRNA expression of ferroptosis-related markers in the cells. RESULTS AND CONCLUSION:(1)Patients were divided into high and low stemness index groups based on the median stemness index.The progression-free survival of patients in the high stemness index group was lower than that in the low stemness index group by disease risk prediction,suggesting poor prognosis.(2)According to GO and KEGG analysis,the groups with high and low stemness indices differed from one another.There were differences in immune infiltration between the high and low stemness index groups.Nine of the 23 ferroptosis-related genes in the differential genes have the potential to establish a highly correlated network of protein interactions.Patients with high expression of IDO1,IFNG,and AQP5 have a better prognosis,while those with high expression of CA9 have a poor prognosis.(3)The qRT-PCR results demonstrated a significant upregulation of stem cell-related markers NANOG,SOX2,and OCT4 mRNA expressions in the stem cell group compared to the adherent cell group(P<0.05).Compared to the adherent cell group,the stem cell group exhibited decreased mRNA expression level of ferroptosis-related marker SLC7A11(P<0.05)while showing increased levels of ACSL4,GPX4,FTH1,and COX2(P<0.05).Compared to the adherent cell group,the stem cell group displayed decreased mRNA expression level of differentially expressed gene CA9 alongside elevated levels of IDO1,IFNG,and AQP5(P<0.05).Stem cells were strongly associated with sarcoma survival and ferroptosis by bioinformatics analysis and experimental verification.Sarcoma stem cells have aberrant expression of CA9,IDO1,IFNG,and AQP5,which may serve as new targets for sarcoma therapy as well as diagnostic indicators.
3.Interpretation and thoughts on the formulation and revision of the standards for exogenous harmful residues in traditional Chinese medicinal materials in the Chinese Pharmacopoeia 2025 Edition
WANG Ying ; SHEN Mingrui ; LIU Yuanxi ; ZUO Tiantian ; WANG Dandan ; HE Yi ; CHENG Xianlong ; JIN Hongyu ; LIU Yongli ; WEI Feng ; MA Shuangcheng
Drug Standards of China 2025;26(1):083-092
As people’s attention to health continues to increase, the market demand for traditional Chinese medicine (TCM) is growing steadily. The quality and safety of Chinese medicinal materials have attracted unprecedented social attention. In particular, the issue of exogenous harmful residue pollution in TCM has become a hot topic of concern for both regulatory authorities and society. The Chinese Pharmacopoeia 2025 Edition further refines the detection methods and limit standards for exogenous harmful residues in TCM. This not only reflects China’s high-level emphasis on the quality and safety of TCM but also demonstrates the continuous progress made by China in the field of TCM safety supervision. Basis on this study, by systematically reviewing the development history of the detection standards for exogenous harmful residues in TCM and analyzing the revisions and updates of these detection standards in the Chinese Pharmacopoeia 2025 Edition, deeply explores the key points of the changes in the monitoring standards for exogenous harmful residues in TCM in the Chinese Pharmacopoeia 2025 Edition. Moreover, it interprets the future development directions of the detection of exogenous residues in TCM, aiming to provide a reference for the formulation of TCM safety supervision policies.
4.Protective Effect of Tongluo Baoshen Prescription-containing Serum on Lipopolysaccharide-induced Podocyte Injury of Rats
Yongfang LIU ; Tiantian YIN ; Huiyang LIU ; Rui HUANG ; Zhiying FENG ; Li ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):139-148
ObjectiveTo observe the effects of Tongluo Baoshen prescription (TLBS)-containing serum on the rat podocyte injury induced by lipopolysaccharide (LPS) and explore the potential mechanisms. MethodsSD rats were used to prepare the blank serum, losartan potassium-containing serum, and low-, medium-, and high-dose TLBS-containing sera. Rat podocytes were cultured in vitro, and the effects of drug-containing sera on podocyte viability were detected by the cell counting kit-8 (CKK-8) method. The optimal intervention volume fraction of drug-containing sera and the optimal concentration of LPS for inducing the podocyte injury were determined. Rat podocytes were grouped as follows: normal control (NC, 10% blank serum), model control (MC, 20.00 mg·L-1 LPS+10% black serum), losartan potassium (LP, 20.00 mg·L-1 LPS+10% losartan potassium-containing serum), low-dose TLBS (TLBS-L, 20.00 mg·L-1 LPS+10% low-dose TLBS-containing serum), medium-dose TLBS (TLBS-M, 20.00 mg·L-1 LPS+10% medium-dose TLBS-containing serum), and high-dose TLBS (TLBS-H, 20.00 mg·L-1 LPS+10% high-dose TLBS-containing serum), and the interventions lasted for 48 h. The ultrastructure of podocytes was observed under a transmission electron microscope. The podocyte apoptosis was detected by the terminal deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL) kit. Immunofluorescence was used to detect the expression of gasdermin D N-terminal fragment (GSDMD-NT) in podocytes. The mRNA and protein levels of G protein-coupled receptor family C group 5 member B (GPRC5B), nuclear factor-κB (NF-κB) p50, NF-κB p52, NF-κB p65, Rel B, c-Rel, NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate-specific protease-1 (Caspase-1), GSDMD-NT, interleukin (IL)-1β, IL-18, nephrin, integrin α3, and integrin β1 in podocytes were determined by real-time quaritiative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultsCompared with the NC group, the MC group showed reduced podocyte protrusions and organelles, segmental missing of cell membranes, increased and swollen mitochondria, irregular nuclear membranes, light chromatin, increased TUNEL fluorescence-positive nuclei (P<0.01), obviously enhanced fluorescence intensity of GSDMD-NT, up-regulated mRNA and protein levels of GPRC5B, NF-κB p50, NF-κB p52, NF-κB p65, Rel B, c-Rel, NLRP3, caspase-1, GSDMD-NT, IL-1β, and IL-18 (P<0.01), and down-regulated mRNA and protein levels of nephrin, integrin α3, and integrin β1 (P<0.01) in podocytes. Compared with the MC group, the LP, TLBS-M, and TLBS-H groups showed improved ultrastructure of podocytes with increased protrusions, intact cell membranes, reduced organelles, and alleviated mitochondrial swelling, decreased TUNEL fluorescence-positive nuclei (P<0.01), weakened fluorescence intensity of GSDMD-NT, down-regulated mRNA and protein levels of GPRC5B, NF-κB p50, NF-κB p52, NF-κB p65, Rel B, c-Rel, NLRP3, caspase-1, GSDMD-NT, IL-1β, and IL-18 (P<0.01), and up-regulated mRNA and protein levels of nephrin, integrin α3, and integrin β1 (P<0.05, P<0.01). Moreover, the changes above were the most obvious in the TLBS-H group. ConclusionThe TLBS-containing serum can regulate the GPRC5B/NF-κB/NLRP3 pathway to inhibit pyroptosis, thereby ameliorating the podocyte injury induced by LPS.
5.Influence of occupational hazard factors on incidence of hypertension in calcium carbide plant: Historical cohort study
Meng LIU ; Wei ZHANG ; Qi ZHUANG ; Xingyu PENG ; Jinling LIU ; Tiantian CHEN ; Hongmei GU ; Shu GUO
Journal of Environmental and Occupational Medicine 2025;42(5):565-570
Background The high work intensity and possible subsequently increased susceptibility to occupational hazards of calcium carbide plants may lead to hypertension in workers, but there are few studies on the relationship between occupational hazard exposure and hypertension in workers involving the production process of calcium carbide. Objective To explore the influence of occupational hazards on the incidence of hypertension in calcium carbide plants. Methods Using historical cohort design, the employees of a calcium carbide factory in the western part of Inner Mongolia Autonomous Region were selected as research subjects. According to the pre-determined inclusion and exclusion criteria, the study population comprised an exposure group of 377 employees (including furnace workers, inspection workers, and maintenance workers) exposed to dust, noise & carbon monoxide, and a control group of 388 employees (including central control workers, electricians, and administrative personnel) without above-mentioned exposure. The total sample size was 765 participants. The follow-up period was from April 2011 to October 2022, and the study endpoint was defined as the conclusion of the follow-up period or diagnosed hypertension in annual occupational health examination. Information on general demographic characteristics, living habits, and work status was collected from all study subjects. Cox proportional hazards regression model was used to analyze the association between occupational hazard exposure and the risk of hypertension among the calcium carbide plant employees. Results The average age, mean systolic and diastolic blood pressure, proportion of males, smoking rate, and alcohol consumption rate in the exposure group were higher than those in the control group (P<0.05). Compared to baseline, both systolic and diastolic blood pressure levels increased in the exposure group and the control group at the end of the follow-up (P<0.05). At the end of the follow-up, the average differences between systolic/ diastolic blood pressure and baseline values in the exposure group were higher than those in the control group (P<0.05). During the follow-up period, a total of 223 cases of hypertension occurred, with a total follow-up of
6.Effects of different storage temperatures and durations on the activity of coagulation factor Ⅷ and Ⅸ in whole blood
Hehe WANG ; Tiantian WANG ; Jie WANG ; Cuicui QIAO ; Wei LIU ; Xueqin ZHANG ; Yan CHENG ; Yunhai FANG ; Xinsheng ZHANG
Chinese Journal of Blood Transfusion 2025;38(6):824-827
Objective: To investigate the effects of different storage temperatures and durations on the activities of coagulation factor Ⅷ (Factor Ⅷ, FⅧ) and coagulation factor Ⅸ (Factor Ⅸ, FⅨ) after whole blood collection, so as to provide data support for the optimal storage conditions. Methods: A total of 16 mL of whole blood was collected from each of the 20 healthy volunteers at our blood center and aliquoted into 8 sodium citrate anticoagulant tubes. Two tubes were immediately centrifuged for the measurement of FⅧ and FⅨ activity levels. The remaining 6 tubes of whole blood were respectively stored under room temperature and low-temperature conditions. At 2, 4, and 6 h, the whole blood samples were centrifuged and analyzed for FⅧ and FⅨ activity levels. The mean values of the two immediately tested tubes were used as the control group, while other tubes were designated as the experimental groups for comparison. Statistical analysis was performed using SPSS 26.0. Results: The activity of FⅧ in whole blood remained stable after 4 hours of storage at both room temperature and low temperature (116.53±25.95 vs 125.22±27.33, 109.77±23.23 vs 125.22±27.33) (P>0.05 for both). However, by 6 hours, FⅧ activity showed a statistically significant decline compared to the control group (108.65±22.92 vs 125.22±27.33, 100.46±20.19 vs 125.22±27.33) (P<0.05 for both), though the room temperature group results were closer to the control values. The activity of FⅨ in whole blood remained stable after 6 hours of storage under both conditions (97.14±19.48 vs 96.76±19.67, 97.10±17.45 vs 96.76±19.6) (P>0.05 for all comparisons). Conclusion: For whole blood samples after collection, storage at either room temperature or low temperature for up to 4 hours does not compromise the accuracy of test results. When stored for 6 hours, FⅨ activity remains stable, whereas FⅧ activity decreases significantly. Notably, FⅧ activity demonstrates better stability at room temperature than under low-temperature conditions within the 6-hour storage.
7.Association between Chinese visceral adiposity index and diabetes mellitus and hypertension among elderly people in Hebei Province
Fujuan YUE ; Xiaoli LIU ; Lijuan TANG ; Fan ZHANG ; Yajing CAO ; Tiantian GUO ; Wen LI ; Dongsheng JIANG
Journal of Public Health and Preventive Medicine 2025;36(6):53-57
Objective To investigate the association between the Chinese visceral adiposity index (CVAI) and diabetes mellitus, hypertension, diabetes mellitus or hypertension, and diabetes with hypertension among elderly people in Hebei Province. Methods In 2020, a stratified multi-stage random sampling was used to conduct questionnaire survey, physical examination and laboratory detection among permanent residents of 10 monitoring sites in Hebei Province. Logistic regression was used to analyze the association between CVAI and diabetes mellitus, hypertension, diabetes mellitus or hypertension, and diabetes with hypertension. The area under the ROC curve (AUC) was used to evaluate the predictive value of CVAI for diabetes mellitus, hypertension, diabetes mellitus or hypertension, and diabetes with hypertension. Results The detection rates of diabetes mellitus, hypertension, diabetes mellitus or hypertension, and diabetes with hypertension were 19.8%, 74.6%, 78.2%, and 16.2%, respectively. Multivariate logistic regression analysis showed that compared with the lowest quartile of CVAI group Q1, the OR (95% CI) of diabetes mellitus, hypertension, diabetes mellitus or hypertension, and diabetes with hypertension in the highest quartile Q4 group were 3.55 (2.58~4.89), 2.52 (1.92~3.31), 3.09 (2.31~4.12), and 4.92 (3.40~7.12), respectively. The ROC curve results showed that CVAI had the best predictive value in the diagnosis of diabetes with hypertension, and the optimized critical values in males and females were 128.54 and 141.88, respectively. Conclusion The detection rates of diabetes mellitus and hypertension are high in the elderly population in Hebei Province. CVAI is positively associated with the risk of diabetes mellitus, hypertension, diabetes mellitus or hypertension, and diabetes with hypertension among the elderly in Hebei. CVAI has the strongest prediction ability for diabetes with hypertension.
8.Effects of moxibustion at "Xinshu" (BL15) and "Feishu" (BL13) on myocardial transferrin receptor 1 and ferroptosis suppressor protein 1 in chronic heart failure rats.
Bing GAO ; Pan LIU ; Lan LI ; Tiantian GONG ; Ling ZHU ; Liya LI ; Ran XIA ; Jing WANG
Chinese Acupuncture & Moxibustion 2025;45(6):781-790
OBJECTIVE:
To observe the effects of moxibustion at "Xinshu" (BL15) and "Feishu" (BL13) on myocardial transferrin receptor 1 (TfR1), ferroptosis suppressor protein 1 (FSP1), atrial natriuretic peptide (ANP), and typeⅠcollagen myocardial collagen fibers (CollagenⅠ) in rats with chronic heart failure (CHF), and to explore the mechanism of moxibustion for ameliorating myocardial fibrosis and improving cardiac function in CHF.
METHODS:
Fifty SD rats were randomly divided into a normal group (n=10) and a modeling group (n=40). The CHF model was established in the modeling group by ligating the left anterior descending coronary artery. After successful modeling, the rats were randomly divided into a model group (n=9), a moxibustion group (n=8), a rapamycin (RAPA) group (n=9), and a moxibustion+RAPA group (n=9). In the moxibustion group, moxibustion was delivered at bilateral "Feishu"(BL13) and "Xinshu" (BL15), 15 min at each point in each intervention, once daily, for 4 consecutive weeks. In the RAPA group, RAPA solution was administered intraperitoneally at a dose of 1 mg/kg, once daily for 4 consecutive weeks. In the moxibustion+RAPA group, RAPA solution was administered intraperitoneally after moxibustion. Ejection fraction (EF) and left ventricular fractional shortening (FS) were measured after modeling and intervention. After intervention, morphology of cardiac muscle was observed using HE staining and Masson's trichrome staining. Total iron content in myocardial tissue was detected using a colorimetric method. Western blot and qPCR were adopted to detect the protein and mRNA expression of TfR1, FSP1, ANP, and CollagenⅠ in myocardial tissue.
RESULTS:
Compared with the normal group, the EF and FS values decreased (P<0.01); necrosis, edema, degeneration, and arrangement disorder were presented in cardiomyocytes; inflammatory cells were obviously infiltrated, the structure of myocardial fibers was disarranged, the collagen fibers were obviously deposited and fibrosis increased (P<0.01); the total iron content and the protein and mRNA expression of TfR1, ANP, and CollagenⅠ in myocardial tissue were elevated (P<0.01), while the protein and mRNA expression of FSP1 were reduced (P<0.01) in the model group. Compared with the model group, the moxibustion group showed that EF and FS increased (P<0.01); myocardial cell morphology was improved, and myocardial fibrosis was alleviated (P<0.01); the total iron content and the protein and mRNA expression of TfR1, ANP, and CollagenⅠ in myocardial tissue decreased (P<0.01), while the protein and mRNA expression of FSP1 increased (P<0.01, P<0.05). Compared with the model group, the myocardial fibrosis was increased (P<0.05); the total iron content and the protein and mRNA expression of TfR1, ANP, CollagenⅠ in myocardial tissue were increased (P<0.01), while protein and mRNA expression of FSP1 decreased (P<0.01) in the RAPA group. When compared with the RAPA group and the moxibustion + RAPA group, EF and FS were elevated (P<0.01, P<0.05); myocardial cells were improved in morphology, the total iron content and the protein and mRNA expression of TfR1, ANP, and CollagenⅠ in myocardial tissue decreased (P<0.01), while protein and mRNA expression of FSP1 increased (P<0.01) in the moxibustion group. In comparison with the moxibustion + RAPA group, the RAPA group showed the decrease in EF and FS (P<0.01), the worsened myocardial fibrosis (P<0.01), the increase in the total iron content and the protein and mRNA expression of TfR1, ANP, and CollagenⅠ in myocardial tissue (P<0.01), and the decrease in the protein and mRNA expression of FSP1 (P<0.01).
CONCLUSION
Moxibustion at "Feishu" (BL13) and "Xinshu" (BL15) can slow down the process of myocardial fibrosis and improve cardiac function in CHF rats. The mechanism of moxibustion may be related to inhibiting ferroptosis through regulating autophagy.
Animals
;
Rats
;
Heart Failure/physiopathology*
;
Moxibustion
;
Rats, Sprague-Dawley
;
Male
;
Receptors, Transferrin/genetics*
;
Myocardium/metabolism*
;
Acupuncture Points
;
Humans
;
Chronic Disease/therapy*
;
Antigens, CD/metabolism*
9.Treatment of irreducible intertrochanteric femoral fractures in elderly by novel bone hook combined with finger-guided technique.
Zenghui ZHANG ; Tichao HAN ; Wei LI ; Yangyang ZHOU ; Junjun LIU ; Nannan LI ; Tiantian REN
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(1):53-58
OBJECTIVE:
To investigate the feasibility and effectiveness of the novel bone hook combined with finger-guided technique in the treatment of irreducible intertrochanteric femoral fractures in elderly.
METHODS:
Between January 2021 and August 2023, 23 elderly patients with irreducible intertrochanteric femoral fractures were treated with the novel bone hook combined with finger-guided technique. There were 10 males and 13 females; the age ranged from 68 to 93 years (mean, 76.2 years). The time from injury to operation ranged from 36 to 76 hours (mean, 51.2 hours). According to the classification standard proposed by TONG Dake et alin 2021, there were 10 cases of typeⅠA, 1 case of typeⅠB, 6 cases of type ⅡA, 4 cases of type ⅡB, and 2 cases of type ⅡC. The operation time, intraoperative blood loss, intraoperative fluoroscopy frequences, and quality of fracture reduction were recorded. The fracture healing time and occurrence of postoperative complications were observed during follow-up. At last follow-up, the Harris scoring system was used to evaluate the hip joint function.
RESULTS:
The operation time was 42-95 minutes (mean, 52.1 minutes). The intraoperative blood loss was 40-420 mL (mean, 126.5 mL). Intraoperative fluoroscopy was performed 14-34 times (mean, 20.7 times). According to the criteria proposed by Chang et al, the quality of fracture reduction was rated as good in 20 cases and acceptable in 3 cases. All patients were followed up 6-20 months (mean, 10.2 months). X-ray film showed that all fractures healed with the healing time of 3.0-5.5 months (mean, 4.0 months). At last follow-up, the Harris score of the hip joint ranged from 82 to 97 points (mean, 90.4 points). Among them, 14 cases were rated as excellent and 9 cases as good. No complication such as coxa vara, cutting of the cephalomedullary nail, nail withdrawal, or nail breakage occurred during follow-up.
CONCLUSION
The treatment of elderly patients with irreducible intertrochanteric femoral fractures by using the novel bone hook combined with finger-guided technique can achieve high-quality fracture reduction and fixation, and has a good effectiveness.
Humans
;
Male
;
Female
;
Aged
;
Aged, 80 and over
;
Hip Fractures/diagnostic imaging*
;
Fracture Fixation, Internal/instrumentation*
;
Fracture Healing
;
Treatment Outcome
;
Operative Time
;
Fracture Fixation, Intramedullary/instrumentation*
;
Bone Nails
;
Postoperative Complications/epidemiology*
;
Feasibility Studies
;
Fingers
10.Mechanism of 4-methylcatechol in inhibiting fibroblast-like synoviocyte migration and suppressing inflammatory responses in treatment of rheumatoid arthritis.
Zhendong YING ; Peng WANG ; Lei ZHANG ; Dailing CHEN ; Qiuru WANG ; Qibin LIU ; Tiantian TANG ; Changjun CHEN ; Qingwei MA
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(8):1051-1060
OBJECTIVE:
To investigate the effects of 4-methylcatechol (4MC) on the migration and inflammatory response in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS), as well as its underlying mechanisms of action.
METHODS:
RA-FLS was isolated from synovial tissue donated by RA patients, and the optimal concentration of 4MC was determined by cell counting kit 8 method for subsequent experiments, and the effect of 4MC on the migratory ability of RA-FLS was evaluated via a cell scratch assay. An inflammation model of RA-FLS was induced by tumor necrosis factor α (TNF-α). Real-time fluorescence quantitative PCR and ELISA were employed to detect the gene and protein expression levels of interleukin-1β (IL-1β) and IL-6 in RA-FLS and their culture supernatants, respectively, thereby investigating the anti-inflammatory effects of 4MC. Western blot was used to examine the expressions of nuclear factor κB (NF-κB) signaling pathway-related proteins, including inhibitor of NF-κB-α (IKBα), phosphorylated (P)-IκBα, NF-κB-inducing kinase α (IKKα), P-IKKαβ, P-p65, and p65. Cellular immunofluorescence was utilized to detect the expression and localization of p65 in RA-FLS, exploring whether 4MC exerts its anti-inflammatory effects by regulating the NF-κB signaling pathway. Finally, a collagen-induced arthritis (CIA) mouse model was established. The anti-RA effect of 4MC in vivo was evaluated by gross observation and histological examination.
RESULTS:
4MC inhibited RA-FLS migration in a concentration-dependent manner. In the TNF-α-induced RA-FLS inflammation model, 4MC significantly decreased the gene and protein expression levels of IL-1β and IL-6. Furthermore, 4MC markedly reduced the ratios of P-IΚBα/IΚBα, P-IKKαβ/IKKα, and P-p65/p65, thereby blocking the transcriptional activity of p65 by inhibiting its nuclear translocation. This mechanism effectively suppressed the activation of the TNF-α-mediated NF-κB signaling pathway. Animal studies demonstrated that 4MC [10 mg/(kg·day)] significantly lowered serum levels of IL-1β, IL-6, and TNF-α, and alleviated arthritis severity and bone destruction in CIA mice.
CONCLUSION
4MC not only inhibits the migration of RA-FLS but also mitigates their inflammatory response by suppressing the NF-κB signaling pathway, thereby effectively exerting its anti-RA effects.
Synoviocytes/metabolism*
;
Arthritis, Rheumatoid/metabolism*
;
Animals
;
Cell Movement/drug effects*
;
Humans
;
Catechols/therapeutic use*
;
Fibroblasts/drug effects*
;
Mice
;
Tumor Necrosis Factor-alpha/pharmacology*
;
Interleukin-1beta/metabolism*
;
Interleukin-6/metabolism*
;
Signal Transduction/drug effects*
;
NF-kappa B/metabolism*
;
Transcription Factor RelA/metabolism*
;
Synovial Membrane/cytology*
;
Cells, Cultured
;
Male
;
Arthritis, Experimental
;
Anti-Inflammatory Agents/pharmacology*
;
NF-KappaB Inhibitor alpha
;
Inflammation


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