1.Mechanisms of Tianma Goutengyin in Alleviating Neuronal Injury in Vascular Dementia Model Rats by Inhibiting A1 Astrocyte Activation via Regulating TNF-α/STAT3/α1ACT Signaling Pathway
Xiaoyan WANG ; Min ZHAO ; Feng TIAN ; Min XIAO ; Nan QU ; Fugui LIU ; Chixiao LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):56-65
ObjectiveTo investigate the effects of Tianma Goutengyin on the tumor necrosis factor-α (TNF-α)/signal transducer and activator of transcription 3 (STAT3)/α1-antichymotrypsin C-terminal tail fragment (α1ACT) signaling pathway and A1-type astrocytes in a rat model of vascular dementia. MethodsSeventy-two male Sprague-Dawley rats were randomly divided into six groups (n=12 per group): Sham-operated group, model group, Tianma Goutengyin high-, medium-, and low-dose groups (5.13, 10.26, and 20.52 g·kg-1), and a nimodipine group (8.1 mg·kg-1). The vascular dementia model was established by permanent bilateral common carotid artery occlusion, followed by 4 weeks of intervention. Learning and memory ability were evaluated using the novel object recognition test, and behavioral performance was assessed using the forced swimming test. Levels of interleukin-6 (IL-6) and C-C motif chemokine ligand 2 (CCL2) in hippocampal tissue were measured by enzyme-linked immunosorbent assay (ELISA). Hippocampal neuronal morphology was observed by Nissl staining, and apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Immunohistochemistry was used to detect positive expression of brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), and myelin basic protein (MBP). Western blot analysis was performed to measure the protein expression levels of TNF-α, TNF receptor 1 (TNFR1), phosphorylated STAT3 (p-STAT3), α1ACT, IL-6, complement component 3 (C3), BDNF, S100 calcium-binding protein A10 (S100A10), and GFAP in hippocampal tissue. ResultsCompared with the sham-operated group, the model group showed a significantly reduced relative recognition index in the novel object recognition test (P<0.01), prolonged immobility time and increased immobility frequency in the forced swimming test (P<0.01). Hippocampal IL-6 and CCL2 levels were significantly increased (P<0.01). Nissl staining revealed a marked reduction in neuronal number and loss of Nissl bodies (P<0.01). MBP-positive expression was significantly decreased (P<0.01), apoptosis was significantly increased (P<0.01), BDNF-positive expression was significantly reduced (P<0.05), and GFAP-positive expression was significantly increased (P<0.01). In addition, the protein expression levels of TNF-α, TNFR1, p-STAT3, α1ACT, IL-6, and C3 were significantly elevated (P<0.01), while BDNF and S100A10 expression levels were significantly decreased (P<0.01). Compared with the model group, all Tianma Gouteng yin dose groups exhibited a significant increase in the relative recognition index (P<0.05), shortened immobility time and reduced immobility frequency (P<0.05, P<0.01). IL-6 and CCL2 levels were significantly decreased (P<0.01), neuronal number was significantly increased (P<0.05, P<0.01), and MBP-positive expression was significantly enhanced (P<0.01). Apoptosis was significantly reduced (P<0.01), BDNF-positive expression was significantly increased (P<0.05), and GFAP-positive expression was significantly decreased (P<0.01). Moreover, the protein expression levels of TNF-α, TNFR1, p-STAT3, α1ACT, IL-6, and C3 were significantly decreased (P<0.01), while BDNF and S100A10 protein expression levels were significantly increased (P<0.01). ConclusionTianma Goutengyin may inhibit A1-type astrocyte activation in rats with vascular dementia through the TNF-α/STAT3/α1ACT signaling pathway, thereby reducing neuronal apoptosis and improving learning and memory function.
2.Exploration on Scientific Connotation of "Removing Calyx" of Inulae Flos Based on Processing Experience of Old Medicine Workers
Qiao ZHOU ; Zhenni QU ; Menghan LIU ; Lei ZHANG ; Benzheng SU ; Yanpeng DAI ; Dianhua SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):260-266
ObjectiveTo conduct a comparative quality analysis of Inulae Flos, fuzz of Inulae Flos and calyx of Inulae Flos, elucidating the scientific connotation of the "removing calyx" process in the traditional processing of Inulae Flos. MethodsInulae Flos decoction pieces were collected, and the fuzz and calyx of Inulae Flos were prepared according to the experiences of old medicine workers. Subsequently, according to the methods under the "Inulae Flos" item in the 2025 edition of the Pharmacopoeia of the People's Republic of China, the appearance characteristics and thin-layer chromatography(TLC) identification of these samples were tested, and the moisture content, total ash content, extract content were also measured. The characteristic fingerprint patterns of Inulae Flos and fuzz of Inulae Flos were established by high-performance liquid chromatography(HPLC), followed by similarity evaluation, principal component analysis(PCA), and partial least squares-discriminant analysis(PLS-DA). The contents of cryptochlorogenic acid, caffeic acid, 1,3-O-dicaffeoylqunic acid, 1,5-O-dicaffeoylqunic acid, and 1-O-acetyl britannilactone were determined to compare the quality differences of Inulae Flos, fuzz of Inulae Flos, and calyx of Inulae Flos. ResultsThe moisture content of Inulae Flos, fuzz of Inulae Flos, and calyx of Inulae Flos was all<10%. The determination results of total ash content were as follows:Calyx of Inulae Flos>Inulae Flos>fuzz of Inulae Flos, and the determination results of alcohol-soluble extract content were as follows:Fuzz of Inulae Flos>Inulae Flos>calyx of Inulae Flos. HPLC fingerprint patterns of Inulae Flos and fuzz of Inulae Flos were established, and 22 common peaks were identified. The similarity analysis and PCA showed that the overall quality of Inulae Flos and fuzz of Inulae Flos was similar, while the overall quality of calyx of Inulae Flos differed significantly from that of Inulae Flos and fuzz of Inulae Flos. PLS-DA results showed that Inulae Flos, fuzz of Inulae Flos, and calyx of Inulae Flos clustered into distinct groups, indicating significant differences among them. Cryptochlorogenic acid and caffeic acid had relatively high contents in calyx of Inulae Flos, the contents of 1,3-O-dicaffeoylqunic acid and 1,5-O-dicaffeoylqunic acid in Inulae Flos and fuzz of Inulae Flos were higher than those in calyx of Inulae Flos. The order of 1-O-acetyl britannilactone content was determined as follows:fuzz of Inulae Flos>Inulae Flos>calyx of Inulae Flos. ConclusionThe scientific nature of "Removing Calyx" process in the cleansing of Inulae Flos by old medicine workers is demonstrated by the resulting fuzz of Inulae Flos decoction pieces exhibiting enhanced cleanliness and higher content of the index component 1-O-acetyl britannilactone. This study provides a reference basis for further improving and enhancing the processing method and quality control standards of Inulae Flos.
3.Indoor environment management and CO 2 volume concentration of primary and secondary school classrooms in winter across three provinces and municipalities of China
Chinese Journal of School Health 2026;47(2):163-167
Objective:
To evaluate the classroom environmental management and CO 2 volume concentration in primary and secondary schools from Liaoning, Tianjin, and Shanghai, thereby providing a scientific basis for developing targeted strategies to improve classroom air quality.
Methods:
From December 16 to 26, 2024, by using stratified random cluster sampling method, the questionnaire survey was conducted in 72 primary and secondary schools (24 each of primary, junior high, and regular high schools) across Liaoning, Tianjin and Shanghai. Information on heating, ventilation and other classroom environmental management was collected. Additionally, 108 classrooms were selected for on site microclimate measurements, including temperature, humidity, wind speed and CO 2 volume concentration. Univariate analysis and multiple linear regression models were employed to explore related factors of classroom CO 2 volume concentration.
Results:
Among the three provinces/municipalities, 20.8% of schools regularly monitored the microclimate. The overall compliance rate for classroom CO 2 volume concentration was 17.6%. Multiple linear regression analysis showed that CO 2 volume concentration in regular and junior high school classrooms were higher than in primary school classrooms ( β=0.067, 0.046, 95%CI =0.036-0.099, 0.013-0.080); classrooms ventilated regularly in the morning and afternoon had higher CO 2 volume concentration than those ventilated during every break between classes ( β=0.043, 95%CI = 0.007- 0.080); both temperature ( β=0.010, 95%CI =0.004-0.016) and humidity ( β=0.003, 95%CI =0.002-0.004) were positively correlated with CO 2 volume concentration (all P <0.05).
Conclusions
Excessive CO 2 volume concentration in primary and secondary school classrooms is a prominent issue, and ventilation frequency is a key intervenable factor for controlling CO 2 levels. It is recommended to promote ventilation during every break between classes as a core management measure and to emphasize air quality supervision in regular high school classrooms.
4.Exploring the Comorbidity Mechanism,Prevention and Treatment Strategies of Rheumatoid Arthritis with Coronary Atherosclerotic Heart Disease Based on Collateral Disease Theory
Journal of Traditional Chinese Medicine 2026;67(9):951-955
Based on the collateral disease theory, this paper systematically reviews the mechanism laws and the prevention and treatment strategies for the comorbidity of rheumatoid arthritis (RA) and coronary heart disease (CHD). It is believed that the collateral vessels serve as the core pathways connecting the limbs and joints with the zang-fu (脏腑) organs. The comorbidity of RA and CHD follows a progressive transmission pattern from wang bi (尪痹), vessel bi (脉痹) to heart bi (心痹). In this sequence, wang bi refers to damp-heat obstructing the qi collaterals of the limbs and joints. Vessel bi is characterized by the binding of phlegm and stasis, along with hyperactive changes in the collateral pathways, and pathogenic factors penetrating the blood vessels. Heart bi occurs when latent toxins damage the collaterals and lead to failure of heart yang. In terms of prevention and treatment strategies, the principle of "treating disease before it arises" should be adopted as guidance, taking overall therapeutic goal as preventing pathogenic factors from penetrating inward along the collaterals, that is from the body surface and limbs into the zang-fu organs. At the wang bi stage, treatment focuses on clearing heat and draining dampness, diffusing and unblocking qi collaterals, while at the vessel bi stage, it is recommended to clear up phlegm, dissolve stasis, and unblock blood collaterals. At the heart bi stage, treatment should focus on activating blood and unblocking the collaterals, restoring yang and diffusing bi.
5.Exploring the Comorbidity Mechanism,Prevention and Treatment Strategies of Rheumatoid Arthritis with Coronary Atherosclerotic Heart Disease Based on Collateral Disease Theory
Journal of Traditional Chinese Medicine 2026;67(9):951-955
Based on the collateral disease theory, this paper systematically reviews the mechanism laws and the prevention and treatment strategies for the comorbidity of rheumatoid arthritis (RA) and coronary heart disease (CHD). It is believed that the collateral vessels serve as the core pathways connecting the limbs and joints with the zang-fu (脏腑) organs. The comorbidity of RA and CHD follows a progressive transmission pattern from wang bi (尪痹), vessel bi (脉痹) to heart bi (心痹). In this sequence, wang bi refers to damp-heat obstructing the qi collaterals of the limbs and joints. Vessel bi is characterized by the binding of phlegm and stasis, along with hyperactive changes in the collateral pathways, and pathogenic factors penetrating the blood vessels. Heart bi occurs when latent toxins damage the collaterals and lead to failure of heart yang. In terms of prevention and treatment strategies, the principle of "treating disease before it arises" should be adopted as guidance, taking overall therapeutic goal as preventing pathogenic factors from penetrating inward along the collaterals, that is from the body surface and limbs into the zang-fu organs. At the wang bi stage, treatment focuses on clearing heat and draining dampness, diffusing and unblocking qi collaterals, while at the vessel bi stage, it is recommended to clear up phlegm, dissolve stasis, and unblock blood collaterals. At the heart bi stage, treatment should focus on activating blood and unblocking the collaterals, restoring yang and diffusing bi.
6.Diagnosis and differential diagnosis of mucin-rich salivary gland tumors
GUAN Weihang ; LIU Cangwei ; GUO Hao ; LI Jinwei ; WANG Dandan ; QIAO Chunyan ; NIE Mengdong ; QU Ming ; SHI Ce
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(6):606-619
This paper systematically elaborates on the key points of diagnosis and differential diagnosis of salivary gland tumors characterized by a substantial amount of extracellular mucus as a main or prominent feature, and clarifies the core differential features. The term "mucus-rich" specifically denotes that mucus is a major component of the tumor, rather than a focal or minor one. This phenomenon is associated with distinct histogenetic mechanisms: it may result from specific genetic mutations (e.g., AKT1 E17K in mucinous adenocarcinoma) that drive ductal epithelial differentiation into mucus-secreting cells, or from myoepithelial cells secreting glycosaminoglycans that form a myxoid stroma. Salivary gland tumors with abundant extracellular mucus include mucinous cystadenoma, sialadenoma papilliferum-like intraductal papillary tumors, mucinous myoepithelioma, pleomorphic adenoma with mucin-rich stroma, mucinous adenocarcinoma, low-grade mucoepidermoid carcinoma, mucin-rich salivary duct carcinoma and intestinal-type adenocarcinoma. The diagnosis of these tumors is complicated by the dual nature of extracellular mucus: while it is a defining feature of some entities, it can also obscure key diagnostic architectural features in others, leading to histological overlap and inconspicuous diagnostic areas. Given the frequent histological morphological overlap among these tumors, immunohistochemical findings and molecular characteristics have emerged as crucial differential diagnostic criteria. Core differential diagnostic points include the following: histologically, there must be meticulous identification of typical structures obscured by mucin (such as squamoid cells in mucoepidermoid carcinoma and apocrine features in salivary duct carcinoma); in immunohistochemical staining, CK20 is useful for distinguishing intestinal-type adenocarcinoma (positive) from mucinous adenocarcinoma (negative), while androgen receptor aids in differentiating salivary duct carcinoma (positive) from mucoepidermoid carcinoma (negative); and molecular testing plays a critical role in definitive diagnosis (e.g., the AKT1 E17K mutation for mucinous adenocarcinoma, MAML2 rearrangement for mucoepidermoid carcinoma, and MEF2C::SS18 fusion for microsecretory adenocarcinoma). This paper systematically summarizes the core pathological features and differential diagnostic points of mucin-rich salivary gland tumors, aiming to provide a practical reference for clinical pathological diagnosis.
7.Efficacy of audiovisual training on ameliorating core symptoms in children with autism spectrum disorder
QU Zhiyi, LIU Zhao, LI Yi, HE Yingli, CHE Hong, ZHANG Xin
Chinese Journal of School Health 2026;47(5):646-651
Objective:
To explore the effect of a computer assisted audiovisual combined intervention model on the core symptoms of children with autism spectrum disorder (ASD), in order to provide references for enriching intervention and treatment methods for ASD children.
Methods:
From December 2023 to March 2024, 36 ASD children aged 4-8 years were recruited from Tianjin Disabled Persons Rehabilitation Center and Xinxinyu Children s Rehabilitation Center, and were divided into a training group (22 cases) and a control group (14 cases). The training group completed a 12 week audiovisual training course (visual sessions:twice a week, for 50-60 minutes each session; auditory sessions:three times a week, for 15 minutes each time), while the control group received only conventional treatment interventions. Before and after the intervention, the core symptoms of ASD children were assessed using the Short Sensory Profile (SSP), Social Responsiveness Scale (SRS), and Repetitive Behavior Questionnaire-2 ( RBQ- 2). Eye tracking experiments were used to test preferences for social attention.
Results:
Before the intervention, there were no statistically significant differences in the total scores and factor scores of the SSP and RBQ-2 scales between the two groups ( t =-1.63, 0.38, both P >0.05). The SRS total score, social communication, and restricted interests and repetitive behavior factor scores of the training group (90.68±25.83, 33.36±11.80, 15.64±7.00) were significantly higher than those of the control group (72.29±19.84, 24.93±7.85, 10.21±5.67) ( t =2.27, 2.36, 2.43, all P <0.05). Children in the training group with higher social communication factor scores before the intervention scored lower than the control group at the post intervention test (simple slope=-14.17, t =-2.48, P = 0.02), while there was no statistically significant difference in post intervention scores between children with lower social communication factor scores before the intervention and the control group (simple slope=2.31, t =0.57, P >0.05). Eye tracking experiments showed that the total fixation time on geometric images decreased significantly more in the training group [ -4.56 (-11.42, 1.21)] compared to the control group [6.55 (-0.32, 16.53)] after the intervention ( Z=2.48, P <0.05).
Conclusions
The computer assisted audiovisual intervention model can effectively improve the core symptoms of ASD children with poorer social communication levels. The promotion of the intervention model needs to consider individual differences in ASD.
8.Differentiation and Treatment of Impaired Glucose Regulation from the Perspective of "Internal Generation of Turbid Pathogen"
Xuan YANG ; Junlin LIU ; Jiayao QU ; Xiangyu LI ; Xinrong FAN
Journal of Traditional Chinese Medicine 2026;67(11):1220-1224
Impaired glucose regulation (IGR) is an intermediate state between normal blood glucose and diabetes, falling under the category of splenic pure heat in traditional Chinese medicine (TCM). The core pathogenesis is believed to be the dysfunction of the zang-fu (脏腑) organs, leading to metabolic abnormalities of subtle essence and the generation of "turbid pathogen". Its accumulation can block qi movement, damage the vessel collaterals, and promote the disease progression toward diabetes. The concept of "turbid pulse" is proposed as an early sign of IGR, characterized by full and heavy sensation with stagnated and astringent circulation, which can serve as an effective supplement to clinical screening. The differentiation and treatment approach in clinical practice is summarized as follows. If the ascending and descending mechanisms of qi movement in the middle jiao (焦) are disrupted, with mutual interference between the clear and the turbid, treatment should focus on harmonizing the ascending and the descending, and transforming dampness with aromatic medicinals. For spleen-stomach qi deficiency due to long-term restriction of the spleen and stomach's function of transportation and transformation, it is suggested to fortify spleen and harmonize stomach, and transport turbid with sweet-warm medicinals. For spleen deficiency and liver constraint induced by internal generation of damp-heat in the middle jiao, the treatment should focus on soothing the liver and opening constraint, and directing the turbid downward with sour-sweat medicinals. For solid accumulation and stagnation caused by liver qi constraint, it is recommended to unblock intestine and remove stagnation, and discharge turbid with bitter-cold medicinals. When the disease is transmitted to the lower jiao, and the kidneys are affected, the treatment should be regulating spleen and kidney, and rectifying turbid with salty-sour medicinals. In conclusion, it is emphasized to promote the transformation of turbid pathogen to restore metabolic homeostasis, providing methods for the clinical diagnosis and treatment of IGR.
9.Mechanism of electroacupuncture treating detrusor-bladder neck dyssynergia after suprasacral spinal cord injury by proteomics
Liya TANG ; Qirui QU ; Jincan LIU ; Ming XU ; Lu ZHOU ; Qiong LIU ; Kun AI
Digital Chinese Medicine 2025;8(2):267-278
Objectives:
To elucidate the potential mechanisms of electroacupuncture (EA) in restoring detrusor-bladder neck dyssynergia (DBND) following suprasacral spinal cord injury (SSCI).
Methods:
A total of 52 specific pathogen-free (SPF) grade famale Sprague-Dawley (SD) rats (10 – 12 weeks, 250 – 280 g) were randomly assigned to either a sham group (n = 12) or a spinal cord injury model group (n = 40). In the model group, DBND was induced through Hassan Shaker spinal cord transection at T10 level, with 24 rats meeting inclusion criteria and subsequently randomized into DBND group (n = 12) and EA intervention group (DBND + EA group, n = 12). After spinal shock recovery (day 19 after modeling), DBND + EA group received EA treatment at Ciliao (BL32), Zhongji (RN3), and Sanyinjiao (SP6) acupoints for 20 min per session at 10/50 Hz frequencies, once daily for 10 d. Sham and DBND groups received anesthesia only without EA intervention. On day 29 post-modeling, all rats underwent urodynamic assessments, followed by hematoxylin and eosin (HE) staining, tandem mass tag (TMT) proteomics, and Western blot (WB) analysis of detrusor and bladder neck tissues. Differentially expressed proteins (DEPs) were defined as proteins with P < 0.05, unique peptides ≥ 2, and fold change > 1.2 or < 0.83. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed using KOBAS 3.0 (P < 0.01), and protein-protein interaction (PPI) networks were analyzed using Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) 11.5 and Cytoscape 3.9.1.
Results:
Compared with sham group, DBND group showed significantly elevated leak point pressure (LPP) and maximum cystometric capacity (MCC) (both P < 0.01). EA treatment significantly reduced both LPP and MCC compared with DBND group (P < 0.01 and P < 0.05, respectively). HE staining revealed that EA reduced detrusor fibrosis and improved bladder neck inflammation. TMT proteomics identified 30 overlapping DEPs in detrusor and 59 overlapping DEPs in bladder neck when comparing DBND + EA/DBND groups with sham group. In detrusor tissue, KEGG analysis revealed 10 significantly enriched pathways (P < 0.01), including mitogen-activated protein kinase (MAPK) signaling pathway. PPI analysis showed 22 of 30 DEPs were interconnected. In bladder neck tissue, 14 pathways were significantly enriched (P < 0.01), including relaxin signaling pathway, with 51 of 59 DEPs showing interconnections. Both TMT and WB validations demonstrated that compared with sham controls, DBND rats exhibited upregulated collagen type IV alpha 2 chain (Col4a2) and downregulated guanine nucleotide-binding protein G(z) subunit alpha (Gnaz) in detrusor tissue, while EA treatment normalized both proteins (both P < 0.05). In bladder neck tissue, DBND rats showed decreased expression of smoothelin (Smtn) and calcium-activated potassium channel subunit beta-1 (Kcnmb1) compared with sham controls (both P < 0.01), which were both upregulated following EA treatment (P < 0.01 and P < 0.05, respectively).
Conclusion
EA restores detrusor-bladder neck coordination in DBND through dual-target mechanisms. In detrusor tissue, EA modulates contraction via extracellular matrix remodeling, cyclic adenosine monophosphate (cAMP) signaling pathway regulation, and enhanced adenosine triphosphate (ATP) biosynthesis mediated by neurotransmitters. In bladder neck tissue, EA promotes relaxation by maintaining contractile phenotypes, reducing fibrosis, suppressing smooth muscle excitation, and regulating presynaptic neurotransmitter release. These findings provide mechanistic insights into EA's therapeutic role in managing DBND.
10.Mechanism of Huangqi Guizhi Wuwutang in Treatment of Sarcopenia Associated with Rheumatoid Arthritis by Improving Skeletal Muscle Homeostasis Through Regulation of Autophagy
Yakun WAN ; Yuan LIU ; Yuan QU ; Jingyu GUO ; Ting LIU ; Zhihui BAI ; Di ZHANG ; Ping JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):12-23
ObjectiveThis study aims to explore the mechanism of action of Huangqi Guizhi Wuwutang in treating rheumatoid arthritis (RA)-associated sarcopenia by regulating autophagy and improving skeletal muscle homeostasis based on network pharmacology,bioinformatics,machine learning,and animal experiments. MethodsActive ingredients and targets of Huangqi Guizhi Wuwutang were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP),PubChem,and SwissTargetPrediction databases. RA-related datasets were retrieved from the GEO database,and differential genes were screened. Sarcopenia-related targets were searched through GeneCards and the Comparative Toxicology Database (CTD),and autophagy-related gene sets were downloaded from the Human Autophagy Database (HADb). Their intersection was analyzed to identify autophagy-related therapeutic targets,followed by enrichment analysis. A protein-protein interaction (PPI) network was constructed using the STRING database,and key targets were selected using multiple methods. Machine learning was applied to predict models based on the expression profiles of intersecting targets,and nomogram models were constructed based on key targets. Molecular docking of the top four active ingredients with key targets was performed using AutoDockVina. A collagen-induced arthritis (CIA) rat model was established using bovine type Ⅱ collagen,with SD rats divided into groups including a blank group,a model group,and low-,medium-,and high-dose groups of Huangqi Guizhi Wuwutang (2.44,4.88,and 9.76 g·kg-1) and administered for five consecutive weeks. Joint scores and gastrocnemius muscle mass were recorded and analyzed after modeling. Hematoxylin and eosin (HE) staining and Masson's staining were used to observe pathological changes in muscle tissue. Immunofluorescence staining was applied to observe the protein expression levels of myosin heavy chain (MYHC) and insulin-like growth factor-1 (IGF-1) in skeletal muscle. Western blot was used to detect the protein expression levels of autophagy-related proteins ATG5,Beclin1,LC3B,muscle-specific proteins (MuRF1),MaFbx,and MYHC. Real-time quantitative reverse transcription PCR (Real-time PCR) was performed to measure the mRNA expression levels of ATG5,Beclin1,LC3B,MuRF1,MaFbx,and MYHC in muscle tissue. ResultsNetwork pharmacology revealed that Huangqi Guizhi Wuwutang shared 25 common targets with autophagy genes related to RA-associated sarcopenia. The PPI network and machine learning identified six key targets,which were primarily involved in autophagy and inflammatory pathways. Animal experiments showed that compared to the blank group,the model group had significantly higher joint scores (P<0.01) and lower gastrocnemius muscle index (P<0.01). HE staining indicated a significant reduction in the cross-sectional area of gastrocnemius muscle fibers,with notable inflammatory cell infiltration and muscle atrophy in the model group. Masson's staining revealed obvious collagen fiber proliferation and deposition,with significant muscle fibrosis in the model group. The protein and mRNA expression levels of ATG5,Beclin1,LC3B,MuRF1,and MaFbx were significantly increased (P<0.01),while the protein expression of MYHC and IGF1 was significantly downregulated (P<0.01). Compared with the model group,the high-dose group of Huangqi Guizhi Wuwutang showed significantly reduced protein and mRNA expression levels of ATG5,Beclin1,LC3B,MuRF1,and MaFbx (P<0.01) and increased protein expression levels of MYHC and IGF1 (P<0.01). The cross-sectional area of muscle fibers increased,and the muscle cell morphology approached normal. Moreover,pathological abnormalities in the gastrocnemius muscle were significantly improved,with reduced collagen fiber proliferation (P<0.01). ConclusionHuangqi Guizhi Wuwutang can mediate autophagy by regulating the expression of ATG5,Beclin1,LC3B,and IGF1,thereby reducing skeletal muscle catabolism and improving skeletal muscle homeostasis,which contributes to the treatment of RA-associated sarcopenia. The findings provide insight into the mechanisms underlying the effects of Huangqi Guizhi Wuwutang in the treatment of RA-related sarcopenia and offer a reference for its enhanced clinical application.


Result Analysis
Print
Save
E-mail