1.Huangqi Jianzhongtang Regulates Polarization of Macrophages M1/M2 and Improves Fat Consumption in Cancer Cachexia Mice
Zhiyan FANG ; Haiyan ZHU ; Wenying HUAI ; Cong HUANG ; Ruocong YANG ; Haiyan YU ; Tiane ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):61-69
ObjectiveTo investigate the effects of Huangqi Jianzhongtang (HQJZ) on macrophage polarization and fat consumption in cancer cachexia (CC) mice. MethodsUltra-performance liquid chromatography-quadrupole/electrostatic field Orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to control the quality of HQJZ. (1) In vitro experiment: HQJZ-containing serum was prepared, and the optimal concentration was determined by cytotoxicity assay. Mouse monocyte-derived macrophages (RAW264.7) were cultured and randomly divided into six groups, including a blank group, a classically activated macrophages (M1) group, an alternatively activated macrophages (M2) group, a HQJZ + blank group, a HQJZ+M1 group, and a HQJZ + M2 group. The relative expression of macrophage marker genes CD86, inducible nitric oxide synthase (iNOS), CD206, and arginase-1 (Arg1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR ). (2) In vivo experiment: Thirty-two BALB/c mice were randomly divided into a control group, a model group, a medroxyprogesterone acetate (MPA) group, and a HQJZ group. Except for the control group, the other mice were injected with CT-26 colon cancer cells to establish a CC model. Mice in the MPA and HQJZ groups were given MPA (0.13 g·kg-1·d-1) or HQJZ (13.13 g·kg-1·d-1) by gavage, respectively, while mice in the control and model groups were given an equal volume of saline by gavage, with interventions continued for 10 d. Real-time PCR was used to detect the expression of macrophage markers (iNOS, Arg1, CD86, CD206) and fat browning-related genes uncoupling protein 1 (UCP1) and peroxisome proliferator-activated receptor γ (PPARγ) in epididymal adipose tissue. Western blot (WB) was used to detect protein expression levels of UCP1 and PPARγ. Micro-computed tomography (micro-CT) was used to measure residual fat volume, and hematoxylin-eosin (HE) staining was used to assess fat browning and calculate pathological scores. ResultsIn vitro, the dominant effective concentration of HQJZ-containing serum was 12.5%. Real-time PCR results showed that, compared with the blank group, Arg1 expression decreased in the HQJZ+blank group (P<0.05), CD206 showed a downward trend without statistical significance, while iNOS and CD86 expression were significantly increased (P<0.05). Compared with the M1 group, Arg1 and CD206 expression decreased in the HQJZ+M1 group (P<0.05). Compared with the M2 group, CD206 expression decreased in the HQJZ+M2 group (P<0.05), CD86 expression increased significantly (P<0.01). In vivo, Real-time PCR results showed that, compared with the control group, CD86 and CD206 expression levels were significantly increased in the model group (P<0.01). Compared with the model group, CD206 expression in the MPA group was significantly decreased (P<0.01). In the HQJZ group, CD206 was significantly decreased (P<0.01). WB results showed that, compared with the model group, protein expression of UCP1 and PPARγ was significantly reduced in the HQJZ group (P<0.05, P<0.01). micro-CT results showed that the total white fat volume in the HQJZ group was greater than that in the model group (P<0.05). HE staining results showed that pathological scores in the HQJZ group were lower than those in the model group (P<0.05). ConclusionHQJZ may inhibit white adipose tissue browning by promoting macrophage M1 polarization and suppressing M2 polarization, thereby delaying fat consumption in CC mice.
2.Quality Evaluation of Naomaili Granules Based on Multi-component Content Determination and Fingerprint and Screening of Its Anti-neuroinflammatory Substance Basis
Ya WANG ; Yanan KANG ; Bo LIU ; Zimo WANG ; Xuan ZHANG ; Wei LAN ; Wen ZHANG ; Lu YANG ; Yi SUN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):170-178
ObjectiveTo establish an ultra-performance liquid fingerprint and multi-components determination method for Naomaili granules. To evaluate the quality of different batches by chemometrics, and the anti-neuroinflammatory effects of water extract and main components of Naomaili granules were tested in vitro. MethodsThe similarity and common peaks of 27 batches of Naomaili granules were evaluated by using Ultra performance liquid chromatography (UPLC) fingerprint detection. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technology was used to determine the content of the index components in Naomaili granules and to evaluate the quality of different batches of Naomaili granules by chemometrics. LPS-induced BV-2 cell inflammation model was used to investigate the anti-neuroinflammatory effects of the water extract and main components of Naomaili granules. ResultsThe similarity of fingerprints of 27 batches of samples was > 0.90. A total of 32 common peaks were calibrated, and 23 of them were identified and assigned. In 27 batches of Naomaili granules, the mass fractions of 14 components that were stachydrine hydrochloride, leonurine hydrochloride, calycosin-7-O-glucoside, calycosin,tanshinoneⅠ, cryptotanshinone, tanshinoneⅡA, ginsenoside Rb1, notoginsenoside R1, ginsenoside Rg1, paeoniflorin, albiflorin, lactiflorin, and salvianolic acid B were found to be 2.902-3.498, 0.233-0.343, 0.111-0.301, 0.07-0.152, 0.136-0.228, 0.195-0.390, 0.324-0.482, 1.056-1.435, 0.271-0.397, 1.318-1.649, 3.038-4.059, 2.263-3.455, 0.152-0.232, 2.931-3.991 mg∙g-1, respectively. Multivariate statistical analysis showed that paeoniflorin, ginsenoside Rg1, ginsenoside Rb1 and staphylline hydrochloride were quality difference markers to control the stability of the preparation. The results of bioactive experiment showed that the water extract of Naomaili granules and the eight main components with high content in the prescription had a dose-dependent inhibitory effect on the release of NO in the cell supernatant. Among them, salvianolic acid B and ginsenoside Rb1 had strong anti-inflammatory activity, with IC50 values of (36.11±0.15) mg∙L-1 and (27.24±0.54) mg∙L-1, respectively. ConclusionThe quality evaluation method of Naomaili granules established in this study was accurate and reproducible. Four quality difference markers were screened out, and eight key pharmacodynamic substances of Naomaili granules against neuroinflammation were screened out by in vitro cell experiments.
3.Application of deep range imaging-optical coherence tomography combined with IOL Master 500 in measuring choroidal thickness and axial length in pediatric myopic patients
Xiaoli YANG ; Guiyang ZHANG ; Qian YANG ; Shilong TAO
International Eye Science 2026;26(1):125-128
AIM: To investigate the application of deep optical coherence tomography(DRI-OCT)combined with IOL Master 500 in measuring choroidal thickness and axial length(AL)in pediatric myopic patients, and analyze the relationship between choroidal thickness and AL.METHODS: Prospective study. A total of 210 pediatric myopia patients(210 eyes)admitted between August 2021 and August 2024 were enrolled. Based on spherical equivalent(SE)measurements, they were categorized into a low myopia group(-3.00 D
4.Early predictive value of pre-treatment tear inflammatory factor levels in patients with dry eye
Yingxing JIANG ; Yuqiu XU ; Yunyun YANG ; Yu ZHANG
International Eye Science 2026;26(1):157-162
AIM: To investigate the application value of pre-treatment tear inflammatory factor levels in predicting therapeutic efficacy for dry eye patients.METHODS:Prospective controlled observational study. A total of 120 patients with dry eye(240 eyes)admitted to our hospital from November 2022 to March 2024 were included. Before dry eye treatment, the levels of inflammatory factors, including interlukin-4(IL-4), IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, IL-18, IL-1β, interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α), granulocyte-colony stimulating factor(G-CSF), granulocyte-macrophage colony-stimulating factor(GM-CSF), monocyte chemoattractant protein-1(MCP-1)in the tear fluid were detected by ELISA. According to the treatment protocol in the Chinese Expert Consensus on the Treatment of Dry Eye(2020), the patients were given treatments, and the related factors affecting the treatment outcomes of dry eye patients were analyzed.RESULTS:After continuous treatment for 4 wk, all the patients completed follow-up, and they were divided into the markedly effective group(60 patients, 120 eyes)and the ineffective group(60 patients, 120 eyes)based on their therapeutic effects. The markedly effective group had significantly lower pre-treatment levels of IL-6, IL-10, IL-18, IL-1β, and TNF-α than the poor efficacy group(all P<0.05). IL-6(OR=0.994), IL-18(OR=0.998), IL-1β(OR=0.933), and TNF-α(OR=0.998)were independently associated with treatment efficacy(all P<0.05). The nomogram model yielded a C-index of 0.971(95% CI: 0.950-0.993), with calibration curves closely aligned to the ideal curve. The model demonstrated significant predictive value for early therapeutic efficacy(sensitivity=96.67%, specificity=71.67%, cutoff=208, AUC=0.866, 95% CI=0.794-0.952, P<0.001).CONCLUSION:The nomogram model constructed based on the levels of inflammatory factors in dry eye patients before treatment can well predict the treatment effect of patients.
5.Erjingwan Alleviate Inflammatory Response and Apoptosis in Skeletal Muscle Cells of Sarcopenia via SIRT1/Nrf2/HO-1 Signaling Pathway
Long SHI ; Yang LI ; Hongyu YAN ; Tianle ZHOU ; Zhiwen ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):57-66
ObjectiveTo investigate the effects of the classical Chinese medicine compound prescription Erjingwan on the inflammatory response and apoptosis of skeletal muscle cells in a mouse model of sarcopenia and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway. MethodsForty C57/BL6 male mice were randomized into a control group, a model group, and groups with different doses of Erjingwan (8,16,32 g·kg-1). The mouse model of sarcopenia was established by D-gal-induced skeletal muscle senescence. The body weight and grip strength of mice treated with different doses of Erjingwan were examined to evaluate their physiological functions. Hematoxylin-eosin (HE) staining and Masson staining were used to observe the pathological changes and fibrosis in the skeletal muscle of mice. Enzyme-linked immunosorbent assay (ELISA) was adopted to determine the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum samples of mice, and biochemical tests were conducted to quantify the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in the serum. The protein and mRNA levels of SIRT1, Nrf2, B-cell lymphoma (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsAfter 4 weeks of drug intervention, the model group exhibited significant reductions in body weight and grip strength (P0.01) compared with the control group. Compared with the model group, all doses of Erjingwan increased the body weight in mice at week 8 (P0.01) and grip strength from week 6 (P0.01). HE staining revealed clear muscle fiber structure in the control group, muscle fiber rupture and atrophy in the model group, and dose-dependent repair of muscle fiber structure in the Erjingwan groups. Masson staining showed minimal collagen fibers and mild fibrosis in the control group, collagen fiber proliferation and severe fibrosis in the model group, and collagen proliferation with dose-dependent inhibition of fibrosis in the Erjingwan groups. ELISA results showed that serum levels of TNF-α and IL-6 were elevated in the model group compared with those in the control group (P0.01). After intervention, the low-dose Erjingwan group exhibited a decreased TNF-α level (P0.05), while the medium and high-dose groups showed decreases in both TNF-α and IL-6 levels (P0.01). Biochemical assays revealed that the model group had decreased SOD and GSH levels (P0.01) and an increased MDA level (P0.01) compared with the control group. The medium and high-dose Erjingwan groups exhibited increases in SOD and GSH levels (P0.01) and decreases in MDA level (P0.01), compared with the model group. WB and Real-time PCR results showed that compared with the control group, the model group presented down-regulated protein and mRNA levels of SIRT1, Nrf2, HO-1, and Bcl-2 in the muscle tissue (P0.01) and up-regulated protein and mRNA levels of Bax (P0.01). Compared with the model group, Erjingwan at different doses up-regulated the protein levels of SIRT1, Nrf2, HO-1, and Bcl-2 (P0.01) and down-regulated the protein and mRNA levels of Bax (P0.01) in the muscle tissue. Low-dose Erjingwan elevated the mRNA levels of Nrf2 and HO-1 (P0.05, P0.01), and medium and high-dose Erjingwan up-regulated the mRNA levels of SIRT1, Nrf2, HO-1, and Bcl-2 (P0.01). ConclusionErjingwan reduced the content of inflammatory factors in skeletal muscle cells, improved the antioxidant capacity, and attenuated pathological changes and fibrosis in the muscle of the mouse model of sarcopenia by regulating the SIRT1/Nrf2/HO-1 pathway, inflammatory response, and apoptosis network.
6.Mechanisms of Shenqi Wenfei Prescription in Intervening in Chronic Obstructive Pulmonary Disease in Rats Based on ROS/TXNIP/NLRP3 Signaling Pathway
Di WU ; Mengyao SHI ; Lu ZHANG ; Tong LIU ; Jiabing TONG ; Cheng YANG ; Zegeng LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):78-87
ObjectiveTo investigate the effects and underlying mechanisms of Shenqi Wenfei prescription (SQWF) on chronic obstructive pulmonary disease (COPD). MethodsA rat model of COPD with lung Qi deficiency was established using lipopolysaccharide (LPS) combined with cigarette smoke. Forty-eight SD rats were randomly divided into a blank group, a model group, low-, medium-, and high-dose SQWF groups (2.835, 5.67, 11.34 g·kg-1), and a Yupingfeng group (1.35 g·kg-1). Drug administration began on day 29 after modeling and continued for 2 weeks. The general condition of the rats was observed, and the lung function in each group was assessed. Hematoxylin-eosin (HE) staining was used to observe pathological changes in lung tissue. The proportion of inflammatory cells in bronchoalveolar lavage fluid (BALF) was measured. Apoptosis in lung tissue was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining. The release level of lactate dehydrogenase (LDH) in BALF was detected by a microplate assay. Reactive oxygen species (ROS) levels in lung tissue were detected using fluorescent probes. The levels of malondialdehyde (MDA), total superoxide dismutase (SOD), and reduced glutathione (GSH) in BALF were measured by biochemical methods. Ultrastructural changes in lung cells were observed via transmission electron microscopy. Double immunofluorescence staining was performed to detect the expression of thioredoxin-interacting protein (TXNIP) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in lung tissue. Western blot analysis was used to detect the protein expression of TXNIP, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate-specific protease-1 (Caspase-1), Caspase-1 p20, gasdermin D (GSDMD), GSDMD N-terminal active fragment (GSDMD-N), interleukin-1β (IL-1β), and IL-18 in lung tissue. Serum IL-1β and IL-18 levels were measured by ELISA. ResultsCompared with the blank group, the model group showed lassitude, fatigue, tachypnea, and audible phlegm sounds, and lung function significantly declined (P0.01). Pulmonary emphysema and inflammatory cell infiltration were obvious. The level of inflammatory cells in BALF increased significantly (P0.05). The number of TUNEL-positive cells increased (P0.01). Levels of LDH, ROS, and MDA in BALF increased significantly (P0.01), while GSH and SOD activities decreased significantly (P0.01). Lung tissue cells showed irregular morphology, swollen mitochondria, disrupted cell membranes, and abundant vesicles, i.e., pyroptotic bodies. Protein levels of TXNIP, NLRP3, ASC, Caspase-1, Caspase-1 p20, GSDMD, GSDMD-N, IL-1β, and IL-18 in lung tissue were significantly elevated (P0.01), and serum IL-1β and IL-18 levels also increased significantly (P0.01). Compared with the model group, each medication group showed alleviation of qi deficiency symptoms and improved lung function (P0.01). Pulmonary emphysema and inflammatory cell infiltration were reduced. Inflammatory cell levels decreased (P0.05). The number of TUNEL-positive cells decreased significantly (P0.01). Levels of LDH, ROS, and MDA decreased significantly (P0.05), while GSH and SOD activities significantly increased (P0.01). Morphological and structural damage in lung tissue was improved to varying degrees. Protein levels of TXNIP, NLRP3, ASC, Caspase-1, Caspase-1 p20, GSDMD, GSDMD-N, IL-1β, and IL-18 in lung tissue significantly decreased (P0.01), and serum IL-1β and IL-18 levels also decreased significantly (P0.05). ConclusionSQWF can improve lung function and alleviate inflammatory responses in COPD rats. Its mechanism may be related to regulating the ROS/TXNIP/NLRP3 pathway and inhibiting pyroptosis.
7.In Vitro and In Vivo Chemical Composition Analysis of Reference Sample of Jinshui Liujunjian Based on UPLC-Q-TOF-MS/MS
Xinyue YANG ; Huiyu LI ; Yaqi LOU ; Xingxing WANG ; Guifang YU ; Chenfeng ZHANG ; Zhenzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):166-173
ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was used to detect the reference sample solution, plasma, and tissue samples of Jinshui Liujunjian under positive and negative ion modes, respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times, secondary mass spectral fragments, reference standards, and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian, including 47 flavonoids, 5 amino acids, 13 iridoids, 16 triterpenoid saponins, etc., of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components; 50 prototype components were identified in different tissues, among which 13, 10, 7, 21, 11, 6, 14, and 40 prototype components were identified in the heart, liver, spleen, lung, kidney, brain, large intestine, and stomach, respectively. Among them, 7 compounds such as ferulic acid, glycyrrhizic acid, and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian, primarily composed of flavonoids and triterpenoid saponins, along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.
8.Rapid Discrimination of Processing Degree of Wine-processed Chuanxiong Rhizoma Based on Intelligent Sensory Technology and Multivariate Statistical Analysis
Xiaolong ZHANG ; Xiaoni MA ; Xinzhu WANG ; Po HU ; Yang PAN ; Tulin LU ; Guangming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):174-182
ObjectiveTo explore the changes in color, odor and chemical components during wine-processing of Chuanxiong Rhizoma(CR), identify differential markers, and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis(PLS-DA), principal component analysis(PCA), discriminant factor analysis(DFA) and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models, and differential components were screened out based on variable importance in the projection(VIP) value1. Then, high performance liquid chromatography(HPLC) was employed to detect the content changes of four components(ferulic acid, senkyunolide I, senkyunolide A and ligustilide) during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L*(brightness value), a*(red-green value) and b*(yellow-blue value). Based on chromaticity differences, the color reference ranges were established for moderately processed CR, including L* of 46.75-48.24, a* of 5.37-6.07 and b* of 20.32-21.70. In odor analysis, DFA revealed significant differences among processing stages, and 11 odor markers were identified, with four differential markers(4-hydroxy-3-butylphthalide, isopropyl butyrate, L-limonene and 1-methoxyhexane) based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection, with discrimination accuracy rates of 92.4% and 93.272% for color and odor, respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages, and four differential components were screened out, providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.
9.Analysis of Animal Models of Dry Age-related Macular Degeneration Based on Clinical Disease-syndrome Characteristics of Traditional Chinese and Western Medicine
Xiaoyu LI ; Lina LIANG ; Yun GAO ; Jiahao LI ; Jianying YANG ; Xiaoshan ZHANG ; Honghao BI ; Menglu MIAO ; Huiyi GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):191-197
ObjectiveAge-related macular degeneration (AMD) is one of the leading causes of low vision and blindness in people over 50 years old, and dry AMD (dAMD) is one type for which there is currently no clear treatment. On the basis of the diagnosis and clinical characteristics of dAMD in traditional Chinese and Western medicine, this paper evaluated the fitting degrees of existing animal models of dAMD with clinical characteristics according to the evaluation methods of animal models, and put forward suggestions and prospects. MethodsLiterature on animal models of dAMD was searched against database, and the characteristics of the models were assigned according to the diagnosis criteria of diseases and syndromes of traditional Chinese and Western medicine, and the fitting degrees of the models with clinical characteristics were analyzed and evaluated. ResultsAt present, the animal models of dAMD are mainly established targeting complement factors, chemokines, oxidative damage, lipid/glucose metabolism, and natural strains. Most of the models can simulate the major pathological changes of dAMD, showing the fitting degree of 25%-50% with clinical characteristics in Western medicine. However, the evaluation of traditional Chinese medicine (TCM) syndromes, especially the evaluation of secondary syndromes, is missing, and the models present low fitting degrees with the clinical characteristics in TCM. ConclusionExisting animal models of dAMD are mostly established under the guidance of Western diagnostic standards, which reproduce the main disease characteristics of Western medicine and lack observation of TCM syndromes. Future studies can pay attention to the intervention factors and evaluation systems of spleen deficiency Qi deficiency and liver-kidney Yin deficiency syndrome and build the animal model of dAMD with integration of disease and syndrome based on clinical characteristics of traditional Chinese and Western medicine.
10.Erjingwan Alleviate Inflammatory Response and Apoptosis in Skeletal Muscle Cells of Sarcopenia via SIRT1/Nrf2/HO-1 Signaling Pathway
Long SHI ; Yang LI ; Hongyu YAN ; Tianle ZHOU ; Zhiwen ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):57-66
ObjectiveTo investigate the effects of the classical Chinese medicine compound prescription Erjingwan on the inflammatory response and apoptosis of skeletal muscle cells in a mouse model of sarcopenia and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway. MethodsForty C57/BL6 male mice were randomized into a control group, a model group, and groups with different doses of Erjingwan (8,16,32 g·kg-1). The mouse model of sarcopenia was established by D-gal-induced skeletal muscle senescence. The body weight and grip strength of mice treated with different doses of Erjingwan were examined to evaluate their physiological functions. Hematoxylin-eosin (HE) staining and Masson staining were used to observe the pathological changes and fibrosis in the skeletal muscle of mice. Enzyme-linked immunosorbent assay (ELISA) was adopted to determine the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum samples of mice, and biochemical tests were conducted to quantify the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in the serum. The protein and mRNA levels of SIRT1, Nrf2, B-cell lymphoma (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsAfter 4 weeks of drug intervention, the model group exhibited significant reductions in body weight and grip strength (P0.01) compared with the control group. Compared with the model group, all doses of Erjingwan increased the body weight in mice at week 8 (P0.01) and grip strength from week 6 (P0.01). HE staining revealed clear muscle fiber structure in the control group, muscle fiber rupture and atrophy in the model group, and dose-dependent repair of muscle fiber structure in the Erjingwan groups. Masson staining showed minimal collagen fibers and mild fibrosis in the control group, collagen fiber proliferation and severe fibrosis in the model group, and collagen proliferation with dose-dependent inhibition of fibrosis in the Erjingwan groups. ELISA results showed that serum levels of TNF-α and IL-6 were elevated in the model group compared with those in the control group (P0.01). After intervention, the low-dose Erjingwan group exhibited a decreased TNF-α level (P0.05), while the medium and high-dose groups showed decreases in both TNF-α and IL-6 levels (P0.01). Biochemical assays revealed that the model group had decreased SOD and GSH levels (P0.01) and an increased MDA level (P0.01) compared with the control group. The medium and high-dose Erjingwan groups exhibited increases in SOD and GSH levels (P0.01) and decreases in MDA level (P0.01), compared with the model group. WB and Real-time PCR results showed that compared with the control group, the model group presented down-regulated protein and mRNA levels of SIRT1, Nrf2, HO-1, and Bcl-2 in the muscle tissue (P0.01) and up-regulated protein and mRNA levels of Bax (P0.01). Compared with the model group, Erjingwan at different doses up-regulated the protein levels of SIRT1, Nrf2, HO-1, and Bcl-2 (P0.01) and down-regulated the protein and mRNA levels of Bax (P0.01) in the muscle tissue. Low-dose Erjingwan elevated the mRNA levels of Nrf2 and HO-1 (P0.05, P0.01), and medium and high-dose Erjingwan up-regulated the mRNA levels of SIRT1, Nrf2, HO-1, and Bcl-2 (P0.01). ConclusionErjingwan reduced the content of inflammatory factors in skeletal muscle cells, improved the antioxidant capacity, and attenuated pathological changes and fibrosis in the muscle of the mouse model of sarcopenia by regulating the SIRT1/Nrf2/HO-1 pathway, inflammatory response, and apoptosis network.

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