1.Zuoguiwan Mitigates Oxidative Stress in Rat Model of Hyperthyroidism Due to Kidney-Yin Deficiency via DRD4/NOX4 Pathway
Ling LIN ; Qianming LIANG ; Changsheng DENG ; Li RU ; Zhiyong XU ; Chao LI ; Mingshun SHEN ; Yueming YUAN ; Muzi LI ; Lei YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):43-51
ObjectiveTo decipher the mechanism by which Zuoguiwan (ZGW) treat hyperthyroidism in rats with kidney-Yin deficiency based on the dopamine receptor D4 (DRD4)/nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) signaling pathway. MethodsThe rat model of kidney-Yin deficiency was induced by unilateral intramuscular injection of dexamethasone (0.35 mg·kg-1). After successful modeling, the rats were randomized into model, methimazole (positive control, 5 mg·kg-1), low-, medium-, and high-dose (1.85, 3.70, 7.40 g·kg-1, respectively) ZGW, and normal control groups. After 21 days of continuous gavage, the behavioral indexes and body weight changes of rats were evaluated. The pathological changes of the renal tissue were observed by hematoxylin-eosin staining. The serum levels of thyroid hormones [triiodothyronine (T3), thyroxine (T4), thyroid-stimulating hormone (TSH)], renal function indexes [serum creatine (Scr) and blood urea nitrogen (BUN)], energy metabolism markers [cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP)], and oxidative stress-related factors [superoxide dismutase (SOD), malondialdehyde (MDA), and NADPH)] were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was employed to analyze the expression of DRD4, NOX4, mitochondrial respiratory chain complex proteins [NADH:ubiquinone oxidoreductase subunit S4 (NDUFS4) and cytochrome C oxidase subunit 4 (COX4)], and inflammation-related protein [tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), p38 mitogen-activated protein kinase (MAPK)] pathway in the renal tissue. ResultsCompared with the normal group, the model group showed mental malaise, body weight decreases (P<0.01), inflammatory cell infiltration in the renal tissue, a few residual parotid glands in the thyroid, elevations in serum levels of T3, T4, Scr, BUN, cAMP, cAMP/cGMP, MDA, and NADPH (P<0.01), down-regulation in protein levels of TSH, SOD, and DRD4 (P<0.05, P<0.01), and up-regulation in expression of NOX4, p-p38 MAPK/p38 MAPK, and inflammatory factors (P<0.01). Compared with the model group, ZGW increased the body weight (P<0.05, P<0.01), reduced the infiltration of renal interstitial inflammatory cells, restored the thyroid structure and follicle size, lowered the serum levels of T3, T4, Scr, BUN, cAMP, cAMP/cGMP, MDA and NADPH (P<0.05, P<0.01), up-regulated the expression of TSH, SOD and DRD4 (P<0.05, P<0.01), and down-regulated the expression of NOX4, p-p38 MAPK/p38 MAPK, and inflammatory factors (P<0.05, P<0.01). Moreover, high-dose ZGW outperformed methimazole (P<0.05). ConclusionBy activating DRD4, ZGW can inhibit the expression of NOX4 mediated by the p38 MAPK pathway, reduce oxidative stress and inflammatory response, thereby ameliorating the pathological state of hyperthyroidism due to kidney-Yin deficiency. This study provides new molecular mechanism support for the clinical application of ZGW.
2.Construction and Evaluation of "Constitution-disease-syndrome" Trinity Model for Rodents with Qi Deficiency
Yasheng DENG ; Jiang LIN ; Yujiang XI ; Qian ZHOU ; Yanping FAN ; Wenyue LI ; Yonghui LIU ; Zhaobing NI ; Qiu CHEN ; Xi MING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):274-284
The theory of constitution in traditional Chinese medicine (TCM) has emerged as a new discipline in recent years. Constitution plays a vital role in the onset,progression,transformation,and prognosis of diseases. At present,some clinical scholars have adopted a novel diagnostic and treatment model of "constitution differentiation-disease identification-syndrome differentiation",in which constitution is regarded as a core element throughout the diagnostic and therapeutic process. Constitution is closely associated with etiology,onset,pathogenesis,syndrome differentiation,and treatment. Against this background,the construction of animal models based on constitution holds far-reaching significance for advancing clinical research. This paper focuses on the construction and evaluation of rodent models with Qi-deficiency constitution,aiming to explore how to further induce Qi-deficiency syndromes and related disease states on the basis of Qi-deficiency constitution models,thereby developing an integrated animal model that embodies the trinity of "constitution-disease-syndrome". The establishment of this model not only provides a solid experimental foundation for the development of new therapies and drugs in TCM targeting specific constitutions,diseases,and syndromes,but also greatly promotes the modernization and scientific advancement of TCM theory. By comprehensively applying multidisciplinary technologies and methods,the study evaluates the model's validity,reliability,and practicality,with the aim of opening new avenues for future research in TCM and promoting the development of the field.
3.Construction and Evaluation of "Constitution-disease-syndrome" Trinity Model for Rodents with Qi Deficiency
Yasheng DENG ; Jiang LIN ; Yujiang XI ; Qian ZHOU ; Yanping FAN ; Wenyue LI ; Yonghui LIU ; Zhaobing NI ; Qiu CHEN ; Xi MING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):274-284
The theory of constitution in traditional Chinese medicine (TCM) has emerged as a new discipline in recent years. Constitution plays a vital role in the onset,progression,transformation,and prognosis of diseases. At present,some clinical scholars have adopted a novel diagnostic and treatment model of "constitution differentiation-disease identification-syndrome differentiation",in which constitution is regarded as a core element throughout the diagnostic and therapeutic process. Constitution is closely associated with etiology,onset,pathogenesis,syndrome differentiation,and treatment. Against this background,the construction of animal models based on constitution holds far-reaching significance for advancing clinical research. This paper focuses on the construction and evaluation of rodent models with Qi-deficiency constitution,aiming to explore how to further induce Qi-deficiency syndromes and related disease states on the basis of Qi-deficiency constitution models,thereby developing an integrated animal model that embodies the trinity of "constitution-disease-syndrome". The establishment of this model not only provides a solid experimental foundation for the development of new therapies and drugs in TCM targeting specific constitutions,diseases,and syndromes,but also greatly promotes the modernization and scientific advancement of TCM theory. By comprehensively applying multidisciplinary technologies and methods,the study evaluates the model's validity,reliability,and practicality,with the aim of opening new avenues for future research in TCM and promoting the development of the field.
4.Regulatory Effect and Mechanisms of Liuhuang Zhike Prescription on Glycemic Control in db/db Mice via AMPK/Akt/GSK-3β Signaling Pathway
Fangyuan DENG ; Ting WANG ; Qianli KANG ; Hongmei LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):122-132
ObjectiveTo investigate the regulatory effects and underlying mechanisms of Liuhuang Zhike prescription (LHZK) on blood glucose in type 2 diabetic db/db mice based on the AMP-activated protein kinase/protein kinase B/glycogen synthase kinase-3β (AMPK/Akt/GSK-3β) signaling pathway. MethodsDb/db mice were used as the model animals, and db/m mice served as the blank control. Forty db/db mice were randomly divided into the model group, metformin group (0.14 g·kg-1), and low-, medium-, and high-dose (4.11, 8.21, 16.43 g·kg-1) LHZK groups, with 8 mice in each group. The db/db mice in the metformin and LHZK groups were administered the corresponding drugs by gavage, while the blank control and model groups were given distilled water by gavage once daily for 8 consecutive weeks. Food intake, water consumption, body weight, and fasting blood glucose (FBG) were measured weekly. An automatic biochemical analyzer was used to determine glycated serum protein (GSP), serum triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels. Hematoxylin-eosin (HE) staining was used to observe pathological morphological changes in the liver and pancreatic tissues. Oil red O staining was used to assess lipid accumulation in liver tissue. The anthrone colorimetric method was used to determine hepatic glycogen content. Real-time quantitative PCR (Real-time PCR) was used to detect the mRNA expression levels of insulin receptor substrate-1 (IRS-1), Akt2, phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6Pase) in liver tissue. Western blot was used to detect the protein expression of AMPKα, phosphorylated AMPKα (p-AMPKα), GSK-3β, p-GSK-3β, glycogen synthase (GS), and phosphorylated GS (p-GS) in liver tissue. ResultsCompared with the blank control group, the model group showed significantly increased food intake, water consumption, body weight, FBG, and GSP levels (P<0.01). Pancreatic islets exhibited marked parenchymal cell hyperplasia and interstitial inflammatory cell infiltration. Liver tissue showed obvious steatosis, accompanied by a compensatory increase in hepatic glycogen content (P<0.01). Hepatic G6Pase mRNA expression was increased, while IRS-1 and Akt2 mRNA expression levels were significantly decreased (P<0.01). The p-AMPKα/AMPKα protein expression ratio showed a decreasing trend, whereas the p-GSK-3β/GSK-3β and p-GS/GS protein expression ratios were significantly increased (P<0.01). Compared with the model group, food intake and water consumption showed decreasing trends in all treatment groups. Food intake was significantly reduced in the low- and high-dose LHZK groups and in the metformin group (P<0.05, P<0.01), and water consumption was significantly reduced in the low-dose LHZK group and in the metformin group (P<0.05, P<0.01). No statistically significant differences in body weight were observed among the LHZK groups, whereas body weight in the metformin group was significantly increased (P<0.05, P<0.01). FBG showed a decreasing trend in all treatment groups, with significant decreases in the low-dose LHZK group and the metformin group (P<0.05, P<0.01). GSP levels were significantly reduced in the low-dose LHZK group and in the metformin group (P<0.05, P<0.01). Hepatic steatosis and pancreatic pathological injury were alleviated to varying degrees in all treatment groups. Hepatic glycogen content further increased in all treatment groups, with significant increases in the medium- and high-dose LHZK groups (P<0.05). Real-time PCR results showed that all treatment groups downregulated the mRNA expression of G6Pase and PEPCK in the liver tissues of db/db mice, with significant downregulation of PEPCK mRNA in the low-dose LHZK and metformin groups (P<0.01). Meanwhile, all treatment groups upregulated IRS-1 and Akt2 mRNA expression, with the most pronounced upregulation observed in the medium-dose LHZK group (P<0.01). The p-AMPKα/AMPKα protein expression ratio was significantly increased in the low- and medium-dose LHZK groups (P<0.01). The p-GSK-3β/GSK-3β protein expression ratio was significantly increased in all treatment groups (P<0.05, P<0.01), whereas the p-GS/GS protein expression ratio was significantly decreased in all treatment groups (P<0.01). ConclusionLHZK effectively reduces FBG and GSP levels in type 2 diabetic mice and improves hepatic steatosis and pancreatic islet pathological injury. Its hypoglycemic mechanism may be associated with regulation of the AMPK/Akt/GSK-3β signaling pathway and promotion of hepatic glycogen synthesis.
5.Analysis of Variety Characteristics and Patterns of Marketed Traditional Chinese Patent Medicines for Treating Chronic Gastritis
Daiyue DING ; Changyue SONG ; Shuangfei DENG ; Siyu LI ; Xiangying KONG ; Xiaohui SU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):252-260
ObjectiveThis study aims to systematically review the marketed traditional Chinese patent medicines for treating chronic gastritis (CG) in China. By analyzing their variety characteristics and prescription patterns, it seeks to provide references for clinical syndrome differentiation-based drug selection, treatment method optimization, and the design of high-quality clinical research. MethodsInformation on marketed traditional Chinese patent medicines for treating CG was collected. Microsoft Excel software was used to collate and statistically analyze representative drugs for each pathological stage, market status, syndrome types, and other contents. The Ancient and Modern Medical Case Cloud Platform (V2.3.9) was employed to analyze the formula composition patterns of standardized prescriptions. ResultsA total of 141 marketed traditional Chinese patent medicines for treating CG in China were included. Based on the disease's pathological progression, they can be classified into drugs for non-atrophic gastritis, atrophic gastritis, and precancerous lesions. Post-marketing research reveals that relevant evaluation is only conducted on 17 drugs, of which 2 involve pharmacoeconomic studies and 14 possess standardized evidence-based evidence. The primary dosage forms were capsules, granules, and tablets. From the 100 prescriptions screened according to inclusion/exclusion criteria, the varieties indicated for the stomach collateral stasis syndrome in atrophic gastritis accounted for the highest proportion. The main efficacy distributions were clearing heat, detoxifying, and relieving pain by promoting Qi circulation. Core drugs included Glycyrrhizae Radix et Rhizome, Paeoniae Radix Alba, and Aucklandiae Radix. Medicinal properties were predominantly warm and neutral. Flavors were mainly bitter, pungent, and sweet. The drugs primarily entered the spleen and stomach meridians. Analysis of the package inserts reveals that 67 products list "contraindications", 110 include "precautions", and 23 explicitly state "adverse reactions". ConclusionTraditional Chinese patent medicines for treating CG hold unique value in clinical practice. However, currently there are challenges such as insufficient clarity in syndrome type descriptions within package inserts and a relative lack of high-level evidence-based medical evidence, as well as pharmacoeconomic evaluations. Future efforts should focus on addressing these shortcomings by advancing research on syndrome characteristics and medication patterns based on syndrome differentiation, systematically conducting pharmacoeconomic evaluations, strengthening the accumulation of high-level evidence-based evidence, and, on this basis, improving patient medication adherence. This will comprehensively enhance the clinical application value and scientific connotation of this category of drugs.
6.The management of blood donors tested reactive to HCV in blood screening based on confirmation of HCV infection
Xuelian DENG ; Liang ZANG ; Xiaofang GONG ; Lei ZHOU ; Xiaochun LIU ; Lin WANG ; Lunan WANG
Chinese Journal of Blood Transfusion 2026;39(4):444-451
Objective: To explore the management of blood donors tested reactive to HCV in blood screening based on confirmation of HCV infection. Methods: Multiple HCV antibody assays, repeating HCV RNA testing, follow-up of blood donors and retesting of archive samples were performed to confirm HCV infection, identify infection status, and exclude false positives in blood donors reactive to HCV in blood screening. Results: From 2011 to 2024, the unqualified rate of HCV detection in blood screening was 2.45‰(2 751/1 122 026). Among these, anti-HCV+-&NAT-accounted for 1.85‰, followed by anti-HCV++ at 0.60‰. The proportion of anti-HCV+-&NAT-and HCV RNA yields was extremely low (0.007‰). The positive rate of anti-HCV+-&NAT-samples tested by electrochemiluminescence method (ELCIA) was approximately 7.5%, differing among reagents (P<0.05). The follow-up of anti-HCV+-&NAT-donors showed that 96.2% (202/210) were false positives, but 51.4% of donors remained anti-HCV+-&NAT-during follow-up. Among them, 8 donors (3.8%) could not be ruled out from HCV infection due to positive retesting by ELCIA. Of the anti-HCV+-&NAT-donors who were reactive at the first follow-up, 86.8% remained anti-HCV+-&NAT-at the second follow-up. The sampling confirmation data showed that all of 260 anti-HCV++ donors were confirmed as anti-HCV positive, and the proportion of false positives or missed detections by NAT was very low. Two occult HBV infections (OBIs) and one HBsAg carrier were identified among the 3 anti-HCV +-&NAT+ donors, and no HCV infection was confirmed in 5 anti-HCV--&HCV RNA + donors. Conclusion: The prevalence of HCV among blood donors in Dalian was about 0.06%, with extremely low proportion of window-period infection and slightly higher proportion of resolved infections than that of current infections. The majority of anti-HCV+-&NAT-were false positive. Blood donors confirmed as false positive should be qualified in blood screening 3 months later before next donation. In order to reduce the false positive results, it was advisable to avoid the same type of supplementary reagents as the initial reagents when performing confirmation.
7.Color Space Method Combined with Chemometrics to Determine Processing Degree of Angelicae Sinensis Radix Carbonisata
Liuying QIN ; Yao HUANG ; Lifan GAN ; Yuanjun LIU ; Congyou DENG ; Dongmei SUN ; Lijin LIANG ; Lin ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):201-210
ObjectiveTo study the changing law of appearance color and physicochemical properties of Angelicae Sinensis Radix Carbonisata(ASRC) during the processing by color space method combined with statistical analysis, so as to provide reference for determining the processing endpoint and evaluating the quality of the decoction pieces. MethodsTaking processing time(4, 8, 12, 16 min) and temperature(180, 200, 220, 240 ℃) as factors, ASRC decoction pieces with different processing degrees were prepared in a completely randomized design. Then, the brightness value(L*), red-green value(a*), yellow-blue value(b*), and total chromaticity value (E*ab) of the decoction pieces were determined by spectrophotometer, the color difference value(ΔE) was calculated, and the data of colorimetric values were analyzed by discriminant analysis. At the same time, the pH, charcoal adsorption, and contents of tannins, 5-hydroxymethylfurfural(5-HMF), tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H and ligustilide of ASRC with different processing degrees were determined by pH meter, ultraviolet and visible spectrophotometry and ultra-high performance liquid chromatography(UPLC). Principal component analysis(PCA) was used to analyze the data of physicochemical indexes, after determining the processing technology of ASRC, the canonical discriminant function was established to distinguish the decoction pieces with different processing degrees, and leave-one-out cross validation was conducted. Finally, Pearson correlation analysis was used to explore the correlation between various physicochemical indexes and chromaticity values. ResultsWith the prolongation of the processing time, L*, a*, b* and E*ab all showed a decreasing trend, and the established discriminant model based on color parameters was able to distinguish ASRC with different processing degrees. The pH showed an increasing trend with the prolongation of processing time, and the charcoal adsorption, and the contents of tannins, 5-HMF, and tryptophan all showed an increasing and then decreasing trend. Among them, the charcoal adsorption, contents of tannin and 5-HMF reached their maximum values successively after processing for 8-12 min. While the contents of chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H and ligustilide decreased with the increase of processing time, with a decrease of 60%-80% at 8 min of processing. Therefore, the optimal processing time should be determined to be 8-12 min. PCA could clearly distinguish ASRC with different processing degrees, while temperature had no significant effect on the processing degree. The 12 batches of process validation results(10 min, 180-240 ℃) showed that except for 3 batches identified as class Ⅱ light charcoal, all other batches were identified as class Ⅲ standard charcoal, and the chromaticity values of each batch of ASRC were within the reference range of class Ⅱ-Ⅲ sample chromaticity values. The correlation analysis showed that the chromaticity values were negatively correlated with pH and charcoal adsorption, and positively correlated with contents of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, and ligustilide. And both pH and charcoal adsorption were negatively correlated with the contents of the above components, but the charcoal adsorption was positively correlated with the content of 5-HMF. ConclusionThe chromaticity values and the contents of various physicochemical indicators of ASRC undergo significant changes with the prolongation of processing time, and there is a general correlation between chromaticity values and various physicochemical indicators. Based on the changes in color and physicochemical indicators, the optimal processing time for ASRC is determined to be 8-12 min. This study reveals the dynamic changes of the relevant indexes in the processing of ASRC, which can provide a reference for the discrimination of the processing degree and the quantitative study of the processing endpoint.
8.Professional self-concept of nurses: the influence of work stressors and related factors
Ji CHEN ; Ziling LUO ; Yue SUN ; Lin MA ; Ji LI ; Xiufang LI ; Xiaorong DENG
Sichuan Mental Health 2025;38(2):178-183
BackgroundThe professional self-concept of nurses is a crucial indicator for their personal growth, development and career planning. Previous studies have shown that work stressors during the internship period may lead to a decrease in the level of professional self-concept among nursing students. Given the existing differences in social roles between nursing students and clinical nurses, the influencing factors of professional self-concept in clinical nurses and its relationship with work stressors require further research. ObjectiveTo explore the relationship between work stressors and professional self-concept among nurses, so as to provide references for improving the level of professional self-concept of clinical nurses. MethodsFrom December 2022 to February 2023, a stratified random sampling method was employed to select 260 in-service nurses from Mianyang Hospital of Traditional Chinese Medicine as the study subjects. Chinese Nurses Stressor Scale (CNSS) and Professional Self-concept of Nurses Instrument (PSCNI) were used for assessment. Pearson correlation analysis was conducted to examine the correlation between CNSS scores and PSCNI scores of nurses. Multiple linear regression analysis was utilized to explore the factors influencing the professional self-concept of nurses. ResultsA total of 238 nurses (91.54%) completed valid questionnaires. PSCNI total score yielded a statistical difference among nurses with different marital statuses (F=8.947, P<0.05). PSCNI total scores were significantly higher in nurses with emergency medical service experience than those without such experience (t=2.208, P<0.05), and were significantly lower in nurses with abnormal physical examination findings in the past year than those without abnormal findings (t=-2.584, P<0.05). Correlation analysis revealed that CNSS total score and subscale scores were negatively correlated with PSCNI total score (r=-0.275~-0.169, P<0.01). Multiple linear regression analysis indicated that work expectation-related stressors, marital status and emergency medical service experience might be influencing factors of their professional self-concept (β=-0.350, 0.345, 0.183, P<0.01). ConclusionNurses' work stressors are closely correlated with their professional self-concept, and high levels of stressors related to nurses' expectations may lead to a decrease in their professional self-concept levels. [Funded by 2021 Research Project of Mianyang Municipal Health Commission (number, 202154)]
9.Total Saponins of Dioscoreae Nipponicae Rhizoma Alleviates Gouty Arthritis by Down-regulating COX-2-mediated M1 Macrophage Reprogramming
Lin HUANG ; Shumin LIU ; Huijuan SUN ; Geyu DENG ; Donghua YU ; Yu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):200-207
ObjectiveTo explore the mechanism of total saponins of Dioscoreae Nipponicae Rhizoma (TSDN) in treating gouty arthritis (GA) by regulating cyclooxygenase-2 (COX-2)-mediated M1 macrophage reprogramming by in vivo and in vitro experiments. MethodsIn vivo experiment: 24 male SD rats were randomly allocated into blank, model (GA), TSDN, and celecoxib groups, with 6 rats in each group. After 7 days of administration, pathological changes in the ankle synovial tissue were observed via hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to quantify the mRNA levels of NOD-like receptor protein 3 (NLRP3) inflammasome, apoptosis-associated speck-like protein (ASC), Caspase-1, COX-2, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the synovial tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the serum levels of inducible nitric oxide synthase (iNOS), IL-1β, CD86, CD80, CD206, and arginase-1 (Arg-1). In vitro experiment: The GA model was established by lipopolysaccharide (LPS) + MSU induction, and the inhibitor concentration was screened by the methyl thiazolyl tetrazolium (MTT) assay. RAW264.7 cells were allocated into blank, model, TSDN, dexamethasone, COX-2 inhibitor (celecoxib), and TSDN + COX-2 inhibitor groups. The levels of iNOS, IL-1β, CD86, CD80, CD206, and Arg-1 in the cell supernatant of each group were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein levels of NLRP3 inflammasome, COX-2, IL-1β, and TNF-α in each group were determined by Real-time PCR and Western blot, respectively. ResultsIn vivo experiment: compared with the model group, TSDN reduced the mRNA levels of NLRP3 inflammasome, COX-2, IL-1β, and TNF-α in the synovial tissue (P<0.05, P<0.01). ELISA results showed that TSDN lowered the serum levels of iNOS, IL-1β, CD86, and CD80 (P<0.01) while increasing the serum levels of CD206 and Arg-1 (P<0.01). In vitro experiment: compared with the model group, TSDN and inhibitor down-regulated the mRNA levels of NLRP3 inflammasome, COX-2, IL-1β, and TNF-α and the protein levels of NLRP3 inflammasome, COX-2, cleaved IL-1β, and TNF-α (P<0.01). Compared with TSDN alone, TSDN + COX-2 inhibitor further reduced the mRNA and protein levels of the markers above (P<0.01). Compared with the model group, TSDN and COX-2 inhibitor decreased the levels of IL-1β, iNOS, CD80, and CD86 (P<0.01) and increased the levels of CD206 and Arg-1 (P<0.01) in cells. Compared with TSDN alone, TSDN + COX-2 inhibitor reduced IL-1β, iNOS, CD80, and CD86 levels (P<0.05, P<0.01) and elevated CD206 and Arg-1 levels (P<0.01) in cells. ConclusionTSDN can alleviate GA by downregulating COX-2-mediated M1 macrophage reprogramming and suppressing the inflammatory factors.
10.Total Saponins of Dioscoreae Nipponicae Rhizoma Alleviates Gouty Arthritis by Down-regulating COX-2-mediated M1 Macrophage Reprogramming
Lin HUANG ; Shumin LIU ; Huijuan SUN ; Geyu DENG ; Donghua YU ; Yu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):200-207
ObjectiveTo explore the mechanism of total saponins of Dioscoreae Nipponicae Rhizoma (TSDN) in treating gouty arthritis (GA) by regulating cyclooxygenase-2 (COX-2)-mediated M1 macrophage reprogramming by in vivo and in vitro experiments. MethodsIn vivo experiment: 24 male SD rats were randomly allocated into blank, model (GA), TSDN, and celecoxib groups, with 6 rats in each group. After 7 days of administration, pathological changes in the ankle synovial tissue were observed via hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to quantify the mRNA levels of NOD-like receptor protein 3 (NLRP3) inflammasome, apoptosis-associated speck-like protein (ASC), Caspase-1, COX-2, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the synovial tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the serum levels of inducible nitric oxide synthase (iNOS), IL-1β, CD86, CD80, CD206, and arginase-1 (Arg-1). In vitro experiment: The GA model was established by lipopolysaccharide (LPS) + MSU induction, and the inhibitor concentration was screened by the methyl thiazolyl tetrazolium (MTT) assay. RAW264.7 cells were allocated into blank, model, TSDN, dexamethasone, COX-2 inhibitor (celecoxib), and TSDN + COX-2 inhibitor groups. The levels of iNOS, IL-1β, CD86, CD80, CD206, and Arg-1 in the cell supernatant of each group were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein levels of NLRP3 inflammasome, COX-2, IL-1β, and TNF-α in each group were determined by Real-time PCR and Western blot, respectively. ResultsIn vivo experiment: compared with the model group, TSDN reduced the mRNA levels of NLRP3 inflammasome, COX-2, IL-1β, and TNF-α in the synovial tissue (P<0.05, P<0.01). ELISA results showed that TSDN lowered the serum levels of iNOS, IL-1β, CD86, and CD80 (P<0.01) while increasing the serum levels of CD206 and Arg-1 (P<0.01). In vitro experiment: compared with the model group, TSDN and inhibitor down-regulated the mRNA levels of NLRP3 inflammasome, COX-2, IL-1β, and TNF-α and the protein levels of NLRP3 inflammasome, COX-2, cleaved IL-1β, and TNF-α (P<0.01). Compared with TSDN alone, TSDN + COX-2 inhibitor further reduced the mRNA and protein levels of the markers above (P<0.01). Compared with the model group, TSDN and COX-2 inhibitor decreased the levels of IL-1β, iNOS, CD80, and CD86 (P<0.01) and increased the levels of CD206 and Arg-1 (P<0.01) in cells. Compared with TSDN alone, TSDN + COX-2 inhibitor reduced IL-1β, iNOS, CD80, and CD86 levels (P<0.05, P<0.01) and elevated CD206 and Arg-1 levels (P<0.01) in cells. ConclusionTSDN can alleviate GA by downregulating COX-2-mediated M1 macrophage reprogramming and suppressing the inflammatory factors.

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