1.Evaluation of the comprehensive intervention effect on lunch for primary and secondary school students in Minhang District of Shanghai
HU Yuhuan, ZANG Jiajie, XU Huilin, GUO Qi, HAN Yan, TANG Hongmei, YING Fangjia, LIANG Hao
Chinese Journal of School Health 2025;46(2):191-195
		                        		
		                        			Objective:
		                        			To evaluate the comprehensive intervention effect of lunch for primary and secondary school students in Minhang District, so as to provide a theoretical and practical basis for lunch intervention in school.
		                        		
		                        			Methods:
		                        			From October to December 2023, a convenience sampling method was used to select 1 937 students from one primary and secondary school in Minhang District.A comprehensive intervention measure focusing on "reducing oil and salt" for lunch recipe optimization and nutrition education was carried out, and a questionnaire survey was conducted to evaluate the intervention effect three months later.  Chi square test and Wilcoxon rank test were used to compare the data before and after the intervention.
		                        		
		                        			Results:
		                        			After intervention, the use of cooking oil and salt, the supply of protein and fat in primary and secondary school lunches were reduced, and had no obvious impact on energy and other major nutrients. After intervention, compared to before intervention, the proportion of primary school students who felt that lunch was greasy decreased (8.9%, 6.2%,  χ 2=4.35), and the proportion of primary and secondary school students who felt that lunch were delicious decreased significantly (33.2%, 23.2%; 63.9%, 53.5%,  χ 2=26.39, 17.52) ( P < 0.05 ). Secondary school students also felt reduced variety of food ingredients (46.9%, 38.3%,  χ 2=16.05,  P <0.05). In addition, after intervention, the total surplus rate of primary school students  meals decreased (7.4%, 4.4%,  χ 2=5.73), mainly reflected in the decrease of the surplus rate of staple foods (7.1%, 2.4%,  χ 2=17.39), while the surplus rate of vegetable dishes increased ( 16.0 %, 21.2%,  χ 2=6.01) ( P <0.05). Although there was no significant change in the total surplus rate of meals for secondary school students, the surplus rate of staple foods decreased (12.9%, 5.4%,  χ 2=33.52), while the surplus rates of meat and vegetable dishes increased (11.2%, 26.9%; 17.5%, 33.2%,  χ 2=74.26, 61.88) ( P <0.05). After intervention, there was no statistically significant difference in the overweight and obesity rates of primary school students ( χ 2=0.11,0.43) and secondary school students ( χ 2=0.01,0.00) compared to before intervention( P >0.05). After intervention, the lung capacity of primary school students [1 564 (1 269,1 890) mL] and sitting forward flexion [11.3 (7.6, 15.2) cm] increased compared to before intervention [1 522 (1 259, 1 819 ) mL, 10.5 (6.3, 13.5) cm] ( Z =2.20, 4.68,  P <0.01), but there was no statistically significant difference in lung capacity and sitting forward flexion of secondary school students before and after intervention ( Z =-0.46, -0.08,  P >0.05).
		                        		
		                        			Conclusion
		                        			The comprehensive intervention of school lunch has promoted a significant decrease in the use of oil and salt in lunch and improved the quality of recipes, and has a positive impact on the situation of leftover lunch and the health of students to a certain extent.
		                        		
		                        		
		                        		
		                        	
2.Construction and application effect of “internet+”Tibetan-language medication service platform
Man LIU ; Liang YANG ; Linling WANG ; Yaqing OU ; Ling CHENG ; Liangfen WANG ; Yingqiang WANG ; Xiaoting TANG ; Rong CHEN
China Pharmacy 2025;36(12):1515-1519
		                        		
		                        			
		                        			OBJECTIVE To build a Tibetan-language medication service platform based on “internet+” and evaluate its effect on improving medication compliance and safety of Tibetan patients with chronic disease. METHODS Medication guidance contents of commonly used drugs in the outpatient department were summarized, translated and recorded in Tibetan-language or video to form a “text-audio-video” multi-dimensional “internet+ ” Tibetan-language medication service platform. A total of 387 Tibetan outpatients with chronic disease in our hospital after the implementation of “internet+” Tibetan-language medication service platform (from January 2024 to June 2024) in our hospital were selected as the intervention group, and 387 Tibetan outpatients before the implementation (from January 2023 to June 2023) were selected as the control group. Patients in the control group received conventional window-based Chinese-language medication services, while patients in the intervention group received both conventional window-based Chinese-language medication service and “internet+ ” Tibetan-language medication service. The medication compliance of patients was evaluated using the 12-item Medication Compliance Scale. A six-level causality assessment was conducted as the principles for analyzing adverse drug reactions (ADR) set by the National Center for ADR Monitoring. Additionally, statistics were compiled on the occurrence of ADR that were assessed as “definite”“probable” or “possible” in the causality assessment. RESULTS The proportion (31.0%) of patients with good medication compliance and compliance scores [39.0 (37.0,42.0)] of patients in the intervention group were significantly better than control group [7.0%, 21.0(19.0, 23.0)]( P<0.05). There were no statistically significant differences in the incidence of various types of ADR or the overall incidence between the two groups (P>0.05). CONCLUSIONS The “internet+” Tibetan-language medication service platform is constructed successfully; the service can effectively improve the medication compliance of Tibetan-language patients, but its effect on improving the medication safety of patients is limited.
		                        		
		                        		
		                        		
		                        	
3.Molecular Mechanism Mediated by HIF-1α/HO-1 Signaling Pathway of Guizhi Fulingwan in Suppressing Ferroptosis in Endometriosis
Li TANG ; Yi ZHANG ; Lulu WU ; Yingying LIANG ; Wenying GONG ; Quanning TAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):1-11
		                        		
		                        			
		                        			ObjectiveThis study aims to investigate the molecular mechanism by which Guizhi Fulingwan (GFW) inhibits ferroptosis in endometriosis (EMT) through the regulation of the hypoxia inducible factor-1α/heme oxygenase 1 (HIF-1α/HO-1) signaling pathway. MethodsMachine learning was employed to identify ferroptosis-related biomarkers associated with EMT. Network pharmacology was utilized to identify the active components of GFW and its potential therapeutic targets against EMT, including core targets. Functional enrichment analysis was conducted to explore the biological processes, molecular functions, cellular components, and signaling pathways associated with the potential targets. An EMT rat model was established via autologous transplantation. Thirty female Sprague-Dawley (SD) rats were randomly divided into five groups: sham-operated, model, positive control (dienogest at 0.2 mg·kg-1), low-dose GFW (2.5 g·kg-1), and high-dose GFW (5 g·kg-1). After modeling, the rats received their respective treatment by oral gavage for 28 consecutive days, while the sham and model groups received equal volumes of distilled water. Serum and ectopic endometrial tissues were collected. Hematoxylin and eosin (HE) staining was employed to evaluate morphological alterations in ectopic lesions. Quantitative real-time polymerase chain reaction (Real-time PCR) and Western blot were conducted to assess mRNA and protein expression of HIF-1α, HO-1, glutathione peroxidase 4 (GPX4), spermidine/spermine N1-acetyltransferase (SAT1), and prostaglandin-endoperoxide synthase 2 (PTGS2). Tissue levels of malondialdehyde (MDA), glutathione (GSH), and ferrous iron (Fe²⁺) were quantified using commercial assay kits. Serum levels of interleukin-6 (IL-6) and transforming growth factor-β1 (TGF-β1) were measured via enzyme-linked immunosorbent assay (ELISA). ResultsFive ferroptosis-related biomarkers in EMT were identified: ALOX12, CHAC1, SAT1, AST1, and HO-1. Network pharmacology analysis revealed 42 active components of GFW and 192 potential therapeutic target genes related to EMT treatment, with FOS, JUN, HO-1 identified as core targets. Functional enrichment analysis indicated that the potential targets were primarily involved in oxidative stress response and reactive oxygen species metabolism and were enriched in the HIF-1 signaling pathway. Compared to the sham-operated group, the model group exhibited significant increases in both mRNA and protein expression of HIF-1α, HO-1, and PTGS2, as well as elevated tissue levels of Fe²⁺ and MDA. Conversely, GSH levels and the expression of GPX4 and SAT1 were markedly reduced, and serum levels of IL-6 and TGF-β1 levels were significantly higher (P<0.01). Compared with the model group, all GFW-treated groups showed significant downregulation of HIF-1α and HO-1, reduced Fe²⁺ levels, and downregulated expression of MDA, PTGS2, IL-6, and TGF-β1. Meanwhile, GSH, GPX4, and SAT1 expression levels were significantly increased (P<0.05, P<0.01), effectively ameliorating iron overload and oxidative stress, thereby demonstrating therapeutic efficacy in EMT, with the high-dose GFW demonstrating the most pronounced therapeutic effects. ConclusionGFW exerts therapeutic effects on endometriosis by regulating the HIF-1α/HO-1 signaling pathway to rectify iron metabolism disorders and attenuate free iron-induced oxidative damage. It upregulates the antioxidative defense system to inhibit lipid peroxidation cascades and modulates inflammatory cytokine networks. These effects collectively disrupt the pathological interaction between ferroptosis and chronic inflammation, providing a novel theoretical foundation for the clinical application of GFW in EMT treatment. 
		                        		
		                        		
		                        		
		                        	
4.Correlation between SLC17A1 single nucleotide polymorphism and susceptibility to hyperuricemia in automotive manufacturing workers
Yongxiang TANG ; Yanmei RUAN ; Xiaojuan LIANG ; Yanru LI ; Guanghui DONG
China Occupational Medicine 2025;52(2):129-134
		                        		
		                        			
		                        			Objective To investigate the correlation of solute carrier protein 17 family member 1 (SLC17A1) gene single nucleotide polymorphisms (SNPs) and susceptibility to hyperuricemia (HUA) in automotive manufacturing workers. Methods A total of 192 Han male workers diagnosed with HUA were selected as the case group, 192 Han male workers without HUA from the same enterprises were selected as the control group. These workers were determined by the matching factor of age, total length of service, and body mass index by the 1∶1 case-control study method. Peripheral venous blood from the workers was collected for DNA extraction. Two SNPs of SLC17A1 were genotyped by MassArray system. Results The gene frequency distributions of SLC17A1 rs2096386 and rs1183201 of workers in the control group were in consistent with the Hardy-Weinberg equilibrium test (both P>0.05). The allele frequency distribution of rs2096386, and the genotype and allele frequency distribution of rs1183201 were significantly different between workers in the two groups (all P<0.05). There was no significant difference in genotype frequency distribution of rs2096386 between workers in the two groups (P>0.05). The results of conditional logistic regression analysis showed that workers with G allele at rs2096386 increased the risk of HUA [odds ratio (OR)=1.43, 95% confidence interval (CI)=1.01-2.04], workers with T allele at rs1183201 increased the risk of HUA (OR=2.03, 95%CI =1.29-3.19), after adjusting for confounding factors such as serum creatinine, blood urea nitrogen, alanin aminotransferase and aspartate aminotransferase. While workers with TA and TA+AA genotypes at rs1183201 had a lower risk of HUA than those with TT genotype (OR=0.51, 95%CI =0.30-0.85; OR=0.50, 95%CI =0.30-0.83), workers with TA genotype at rs1183201 had a lower risk of HUA than those with TT+AA genotype (OR=0.53, 95%CI =0.32-0.88). Conclusion The polymorphisms at rs2096386 and rs1183201 of SLC17A1 gene may be correlated with HUA susceptibility among automobile manufacturing workers in Guangzhou City. 
		                        		
		                        		
		                        		
		                        	
5.Study on Kinetic and Static Tasks With Different Resistance Coefficients in Post-stroke Rehabilitation Training Based on Functional Near-infrared Spectroscopy
Ling-Di FU ; Jia-Xuan DOU ; Ting-Ting YING ; Li-Yong YIN ; Min TANG ; Zhen-Hu LIANG
Progress in Biochemistry and Biophysics 2025;52(7):1890-1903
		                        		
		                        			
		                        			ObjectiveFunctional near-infrared spectroscopy (fNIRS), a novel non-invasive technique for monitoring cerebral activity, can be integrated with upper limb rehabilitation robots to facilitate the real-time assessment of neurological rehabilitation outcomes. The rehabilitation robot is designed with 3 training modes: passive, active, and resistance. Among these, the resistance mode has been demonstrated to yield superior rehabilitative outcomes for patients with a certain level of muscle strength. The control modes in the resistance mode can be categorized into dynamic and static control. However, the effects of different control modes in the resistance mode on the motor function of patients with upper limb hemiplegia in stroke remain unclear. Furthermore, the effects of force, an important parameter of different control modes, on the activation of brain regions have rarely been reported. This study investigates the effects of dynamic and static resistance modes under varying resistance levels on cerebral functional alterations during motor rehabilitation in post-stroke patients. MethodsA cohort of 20 stroke patients with upper limb dysfunction was enrolled in the study, completing preparatory adaptive training followed by 3 intensity-level tasks across 2 motor paradigms. The bilateral prefrontal cortices (PFC), bilateral primary motor cortices (M1), bilateral primary somatosensory cortices (S1), and bilateral premotor and supplementary motor cortices (PM) were examined in both the resting and motor training states. The lateralization index (LI), phase locking value (PLV), network metrics were employed to examine cortical activation patterns and topological properties of brain connectivity. ResultsThe data indicated that both dynamic and static modes resulted in significantly greater activation of the contralateral M1 area and the ipsilateral PM area when compared to the resting state. The static patterns demonstrated a more pronounced activation in the contralateral M1 in comparison to the dynamic patterns. The results of brain network analysis revealed significant differences between the dynamic and resting states in the contralateral PFC area and contralateral M1 area (F=4.709, P=0.038), as well as in the contralateral PM area and ipsilateral M1 area (F=4.218, P=0.049). Moreover, the findings indicated a positive correlation between the activation of the M1 region and the increase in force in the dynamic mode, which was reversed in the static mode. ConclusionBoth dynamic and static resistance training modes have been demonstrated to activate the corresponding brain functional regions. Dynamic resistance modes elicit greater oxygen changes and connectivity to the region of interest (ROI) than static resistance modes. Furthermore, the effects of increasing force differ between the two modes. In patients who have suffered a stroke, dynamic modes may have a more pronounced effect on the activation of exercise-related functional brain regions. 
		                        		
		                        		
		                        		
		                        	
6.Efficacy and safety of letermovir in preventing cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation
Ranran WANG ; Shuyue LI ; Ranran LIANG ; Xianmin SONG ; Yuanjun TANG ; Junwei GAO
China Pharmacy 2025;36(15):1904-1909
		                        		
		                        			
		                        			OBJECTIVE To evaluate the efficacy and safety of letermovir in preventing cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS A retrospective cohort study was conducted, enrolling patients who underwent allo-HSCT at the Department of Hematology, Shanghai General Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, from August 30, 2022, to February 21, 2024. Patients who initiated letermovir prophylaxis within 28 days post-transplantation were assigned to the experimental group (99 cases), while those who did not initiate letermovir within this period were assigned to the control group (18 cases). The incidence and clinical characteristics of CMV infection (including the number of wangranran@xinhuamed.com.cn CMV infection cases, the number of cases progressing to CMV disease, recurrent CMV disease, onset time of CMV infection, and treatment duration), immune function recovery within 120 days post-transplantation, and the occurrence of transplantation-related complications (including CD4+ and CD8+ T-cell recovery, Epstein-Barr virus infection, acute graft-versus-host disease, human herpesvirus 6 infection, and posttransplant lymphoproliferative disorders) and adverse events were recorded. Univariate and multivariate Cox regression analyses were performed to identify factors influencing CMV infection. RESULTS A total of 117 patients were included, among whom 15 developed CMV infection, 5 progressed to CMV disease, and 2 experienced recurrent CMV disease. The CMV infection rate in the experimental group was significantly lower than that in the control group (P<0.001), and the onset time of CMV infection was significantly delayed (P=0.014). The proportion of patients with CD4+ T-cell counts ≥200 cells/μL in the experimental group was significantly lower than that in the control group (P=0.022). During the follow-up period, elevated creatinine levels were observed in 1 patient, and nausea and vomiting were observed in 2 patients. Multivariate Cox regression analysis revealed that the use of high-dose corticosteroids was a risk factor for CMV infection (HR=6.230, 95%CI of 1.255-30.926, P=0.025), while initiating letermovir within 28 days post-transplantation was a protective factor (HR=0.125, 95%CI of 0.045-0.348, P<0.001). CONCLUSIONS Early initiation of letermovir after allo-HSCT significantly reduces the CMV infection rate and delays the onset of infection, with favorable short-term safety.
		                        		
		                        		
		                        		
		                        	
7.Textual Research and Ancient and Modern Application of Classical Prescription Sinisan
Lyuyuan LIANG ; Qing TANG ; Jialei CAO ; Wenxi WEI ; Yuxin ZHANG ; Jinyu CHEN ; Hejia WAN ; Chen CHEN ; Ruiting SU ; Bingqi WEI ; Shen'ao DING ; Bingxiang MA ; Wenli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):182-193
		                        		
		                        			
		                        			Sinisan is a classical prescription developed and applied by ancient medical experts and it is first recorded in the Treatise on Cold Damage written by ZHANG Zhongjing in the Eastern Han Dynasty. Later physicians have modified this prescription based on this original one. The bibliometrics methods were used to analyze the key information and research trend of Sinisan. According to the inclusion and exclusion criteria, 69 pieces of effective data were extracted, involving 67 ancient traditional Chinese medicine (TCM) books. The results showed that the name, composition, and decocting methods of Sinisan in later generations were inherited from the original record in the Treatise on Cold Damage. The original plants of medicinal materials used in Sinisan are basically clear. We recommend Bupleuri Radix as the dried root of Bupleurem scorzonerifolium, Paeoniae Radix Alba as the dried root of Paeonia lactiflora, Aurantii Fructus as the dried fruit of Citrus aurantium, Glycyrrhizae Radix et Rhizoma as the dry root and rhizome of Glycyrrhiza uralensis. Raw materials of Bupleuri Radix and Paeoniae Radix Alba, Aurantii Fructus stir-fried with bran, and stir-fried Glycyrrhizae Radix et Rhizoma should be used for preparation of Sinisan. According to measurement system in the Han Dynasty, a bag of Sinisan is composed of 1.25 g Bupleuri Radix, 1.25 g Paeoniae Radix Alba, 1.25 g Aurantii Fructus, and 1.25 g Glycyrrhizae Radix et Rhizoma. The materials should be grounded into coarse powder and taken with a proper amount of rice soup, 3 times a day. Sinisan has the effects of regulating qi movement and harmonizing the liver and spleen. It can be used for treating reversal cold in limbs and cold damage. In modern clinical practice, Sinisan can be used to treat chronic gastritis, irritable bowel syndrome, and dyspepsia. The above research results provide scientific reference for the future research and development of Sinisan. 
		                        		
		                        		
		                        		
		                        	
8.Key Information Research and Ancient and Modern Application Analysis of Classic Prescription Houpo Sanwutang
Wenli SHI ; Qing TANG ; Huimin CHEN ; Jialei CAO ; Bingqi WEI ; Lan LIU ; Keke LIU ; Yun ZHANG ; Yujie CHANG ; Yihan LI ; Jingwen LI ; Bingxiang MA ; Lvyuan LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):214-221
		                        		
		                        			
		                        			Houpo Sanwutang, included in the Catalogue of Ancient Classical Prescriptions (Second Batch), was first recorded in the Synopsis of Golden Chamber written by ZHANG Zhongjing from the Eastern Han dynasty and was modified by successive generations of medical experts. A total of 37 pieces of effective data involving 37 ancient Chinese medical books were retrieved from different databases. Through literature mining, statistical analysis, and data processing, combined with modern articles, this study employed bibliometrics to investigate the historical origin, composition, decoction methods, clinical application, and other key information. The results showed that the medicinal origin of Houpo Sanwutang was clearly documented in classic books. Based on the conversion of the measurements from the Han Dynasty, it is recommended that 110.4 g Magnolia Officinalis Cortex, 55.2 g Rhei Radix et Rhizoma, and 72 g Aurantii Fructus Immaturus should be taken. Magnolia Officinalis Cortex and Aurantii Fructus Immaturus should be decocted with 2 400 mL water first, and 1 000 mL should be taken from the decocted liquid. Following this, Rhei Radix et Rhizoma should be added for further decoction, and then 600 mL should be taken from the decocted liquid. A single dose of administration is 200 mL, and the medication can be stopped when patients restore smooth bowel movement. Houpo Sanwutang has the effect of moving Qi, relieving stuffiness and fullness, removing food stagnation, and regulating bowels. It can be used in treating abdominal distending pain, guarding, constipation, and other diseases with the pathogenesis of stagnated heat and stagnated Qi in the stomach. The above results provide reference for the future development and research of Houpo Sanwutang. 
		                        		
		                        		
		                        		
		                        	
9.Mechanism of imperatorin in ameliorating doxorubicin resistance of breast cancer based on transcriptomics
Yiting LI ; Wei DONG ; Xinli LIANG ; Hu WANG ; Yumei QIU ; Xiaoyun DING ; Hao ZHANG ; Huiyun BAO ; Xianxi LI ; Xilan TANG
China Pharmacy 2025;36(5):529-534
		                        		
		                        			
		                        			OBJECTIVE To investigate the ameliorative effect and potential mechanism of imperatorin (IMP) on doxorubicin (DOX) resistance in breast cancer. METHODS The effects of maximum non-toxic concentration (100 μg/mL) of IMP combined with different concentrations of DOX (12.5, 25, 50, 75, 100 μg/mL) on the proliferation of MCF-7/DOX cells were determined by MTT method. MCF-7/DOX cells were divided into blank control group (1‰ dimethyl sulfoxide), DOX group (50 μg/mL), IMP+DOX group (100 μg/mL IMP+50 μg/mL DOX) and IMP group (100 μg/mL). mRNA and protein expressions of multidrug resistance protein 1 (MDR1) and multidrug resistance-associated protein 1 in each group were measured. The relevant pathways and targets involved in the improvement of DOX resistance in breast cancer cells by IMP were screened and validated by using transcriptome sequencing technology, along with gene ontology (GO) enrichment analyses and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. RESULTS Compared with DOX alone, the combination of IMP and DOX reduced the half inhibitory concentration of DOX on MCF-7/DOX cells from 81.965 μg/mL to 43.170 μg/mL, the reverse fold was 1.90, and the mRNA expression of MDR1 was significantly down-regulated (P<0.05). The results of GO enrichment analyses and KEGG pathway enrichment analyses indicated that the reversal of DOX resistance in breast cancer by IMP was mainly associated with the regulation of biological processes such as detoxification, multiple biological processes, and cell killing. The main pathway involved was the p53 signaling pathway, and the key targets mainly included constitutively photomorphogenic protein 1 (COP1), cyclin E1 (CCNE1), growth arrest and DNA damage-inducible protein 45A E-mail:tangxilan1983@163.com (GADD45A) and GADD45B. The results of the verification experiments showed that compared with DOX group, there was a trend of up-regulation of COP1 mRNA, and significant down- regulation of CCNE1, GADD45A, and GADD45B mRNA expression in IMP+DOX group (P<0.05). CONCLUSIONS The effect of IMP in ameliorating DOX resistance in breast cancer is related to its regulation of COP1, CCNE1, GADD45A and GADD45B targets in the p53 signaling pathway.
		                        		
		                        		
		                        		
		                        	
10.Carnosic acid inhibits osteoclast differentiation by inhibiting mitochondrial activity
Haishan LI ; Yuheng WU ; Zixuan LIANG ; Shiyin ZHANG ; Zhen ZHANG ; Bin MAI ; Wei DENG ; Yongxian LI ; Yongchao TANG ; Shuncong ZHANG ; Kai YUAN
Chinese Journal of Tissue Engineering Research 2025;29(2):245-253
		                        		
		                        			
		                        			BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.
		                        		
		                        		
		                        		
		                        	
            

Result Analysis
Print
Save
E-mail