1.Influence of pterygium thickness and area on corneal refractive status
Xiaodong CHENG ; Jie WANG ; Song GAO ; Yanhong LU ; Yanbo MA ; Xinming CUI ; Xihui CHEN
International Eye Science 2026;26(1):152-156
AIM: To investigate the influence of pterygium thickness and area on corneal refractive status.METHODS: Prospective longitudinal study. A total of 60 cases(60 eyes)of pterygium patients admitted to our hospital from January 2024 to September 2024 were randomly selected. All patients underwent pterygium excision combined with pedicle conjunctival flap transplantation for treatment. Optical coherence tomography(OCT)was used to measure the preoperative thickness of patient's pterygium, and a digital slit lamp microscope was used to measure the area of pterygium. The corneal refractive status(degree of corneal astigmatism and average curvature)and changes in uncorrected visual acuity of patients before surgery, 1 d, 1, and 3 mo after surgery were compared. The relationship between preoperative thickness and area of pterygium in patients and corneal refractive status indicators at different postoperative time points were analyzed, and Logistic regression was used to analyze the impact of pterygium thickness and area on postoperative visual improvement in patients.RESULTS: All patients completed follow-up after surgery for 3 mo. At 3 mo after surgery, visual acuity improved in 21 eyes(35%). The results of bivariate Pearson correlation analysis showed that the thickness and area of pterygium positively correlated with the degree of corneal astigmatism and uncorrected visual acuity before surgery and 1 d, 1, and 3 mo after surgery(all P<0.05), and negatively correlated with the average corneal curvature before surgery and 1 d, 1, and 3 mo after surgery(all P<0.05). Logistic regression analysis showed that the thickness and area of pterygium before surgery, high degree of corneal astigmatism, and low uncorrected visual acuity(large LogMAR value)were all risk factors for poor postoperative visual improvement in patients(OR>1, P<0.05). The large average corneal curvature before surgery was a protective factor for poor postoperative visual improvement in patients(OR<1, P<0.05).CONCLUSION: The increase in thickness and area of pterygium can, to some extent, improve corneal astigmatism, reduce the average curvature of the cornea, and affect postoperative visual recovery.
2.Serological characteristics of individuals with hepatitis C virus/hepatitis B virus overlapping infection
Yanfei CUI ; Xia HUANG ; Chao ZHANG ; Yingjie JI ; Song QING ; Yuanjie FU ; Jing ZHANG ; Li LIU ; Yongqian CHENG
Journal of Clinical Hepatology 2026;42(1):74-79
ObjectiveTo investigate the status of overlapping hepatitis B virus (HBV) infection in patients with chronic hepatitis C virus (HCV) infection and the serological characteristics of such patients. MethodsA total of 8 637 patients with HCV infection who were hospitalized from January 1, 2010 to December 31, 2020 and had complete data of HBV serological markers were enrolled, and the composition ratio of patients with overlapping HBV serological markers was analyzed among the patients with HCV infection. The patients were divided into groups based on age and year of birth, and serological characteristics were analyzed, and the distribution of HBV-related serological characteristics were analyzed across different HCV genotypes. ResultsThe patients with HCV/HBV overlapping infection accounted for 5.85%, and the patients with previous HBV infection accounted for 48.10%; the patients with protective immunity against HBV accounted for 14.67%, while the patients with a lack of protective immunity against HBV accounted for 31.39%. The patients were divided into groups based on age: in the 0 — 17 years group, the patients with protective immunity against HBV accounted for 61.41% (304 patients); the 18 — 44 years group was mainly composed of patients with previous HBV infection (698 patients, 37.31%), the 45 — 59 years group was predominantly composed of patients with previous HBV infection (1 945 patients, 50.38%), and the ≥60 years group was also predominantly composed of patients with previous HBV infection (1 486 patients, 61.66%). The patients were divided into groups based on the year of birth: in the pre-1992 group, the patients with previous HBV infection accounted for 51.63% (4 112 patients); in the 1992 — 2005 group, the patients with protective immunity against HBV accounted for 54.72% (168 patients); in the post-2005 group, the patients with protective immunity against HBV accounted for 64.38% (235 patients). In this study, 6 301 patients underwent HCV genotype testing: the patients with genotype 1b accounted for the highest proportion of 51.71% (3 258 patients), followed by those with genotype 2a (1 769 patients, 28.07%), genotype 3b (63 patients, 1.00%), genotype 3a (10 patients, 0.16%), genotype 4 (21 patients, 0.33%), and genotype 6a (5 patients, 0.08%). ConclusionWith the implementation of hepatitis B planned vaccination program in China, there has been a significant reduction in the proportion of patients with previous HBV infection among the patients with HCV/HBV overlapping infection, but there is still a relatively high proportion of patients with a lack of protective immunity against HBV.
3.Compilation Instruction for Expert Consensus on Clinical Application of Binghuang Fule Ointment
Xin CUI ; Yanping BAI ; Ping SONG ; Yuanyuan LI ; Yanming XIE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):267-273
Compilation instruction for Expert Consensus on Clinical Application of Binghuang Fule Ointment elaborates on the formulation methods and evidence-based basis of the consensus. To address the problems of insufficient evidence on efficacy, vague indications, and a lack of uniform standard for Binghuang Fule Ointment in clinical application, 34 experts from 29 medical institutions across China participated in the compilation under the lead of the Institute of Basic Research in Clinical Medicine and Xiyuan Hospital, China Academy of Chinese Medical Sciences, as well as China-Japan Friendship Hospital. The compilation strictly adhered to the WHO Handbook for Guideline Development (GB/T 1.1—2020), and the Guidance of Instructions for Compiling Expert Consensus on Clinical Practice of Chinese Patent Medicine. Through multidisciplinary collaboration, the compilation was completed using the Grading of Recommendations Assessment, Development and Evaluations (GRADE) evidence grading system. The detailed workflow included various key links. In clinical question construction, 15 items were screened by the nominal group method. In evidence retrieval, Chinese and English databases, along with gray literature, were covered to obtain 116 clinical and 33 pharmaceutical studies. In safety assessment, drug monitoring data and clinical research results were integrated, clarifying local adverse skin reactions and contraindications. Ultimately, 8 recommendations were formed by the GRADE grid method, while 16 consensus suggestions were reached via the majority vote rule. The results showed that the Binghuang Fule Ointment was applicable to eczema, psoriasis, neurodermatitis, tinea pedis, and other diseases. The Consensus also elucidated the syndrome differentiation points, usage and dosage for different diseases (such as adjustment of course and application frequency), as well as the indications of combination medication. Additionally, safety assessment suggested that the Ointment should be used with caution in individuals with skin ulceration or hypersensitivity. To ensure methodological rigor, the compilation process went through three rounds of internal and external expert reviews, while a comprehensive analysis was conducted by literature analysis, the Delphi method, and other methods. This compilation instruction provided methodological support for the clinical transformation of the Consensus through key links, including project initiation, international registration, informed consent, conflict-of-interest statements, evidence evaluation, and popularization. The Consensus will be continuously improved through a dynamic revision mechanism in the future.
4.Compilation Instruction for Expert Consensus on Clinical Application of Binghuang Fule Ointment
Xin CUI ; Yanping BAI ; Ping SONG ; Yuanyuan LI ; Yanming XIE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):267-273
Compilation instruction for Expert Consensus on Clinical Application of Binghuang Fule Ointment elaborates on the formulation methods and evidence-based basis of the consensus. To address the problems of insufficient evidence on efficacy, vague indications, and a lack of uniform standard for Binghuang Fule Ointment in clinical application, 34 experts from 29 medical institutions across China participated in the compilation under the lead of the Institute of Basic Research in Clinical Medicine and Xiyuan Hospital, China Academy of Chinese Medical Sciences, as well as China-Japan Friendship Hospital. The compilation strictly adhered to the WHO Handbook for Guideline Development (GB/T 1.1—2020), and the Guidance of Instructions for Compiling Expert Consensus on Clinical Practice of Chinese Patent Medicine. Through multidisciplinary collaboration, the compilation was completed using the Grading of Recommendations Assessment, Development and Evaluations (GRADE) evidence grading system. The detailed workflow included various key links. In clinical question construction, 15 items were screened by the nominal group method. In evidence retrieval, Chinese and English databases, along with gray literature, were covered to obtain 116 clinical and 33 pharmaceutical studies. In safety assessment, drug monitoring data and clinical research results were integrated, clarifying local adverse skin reactions and contraindications. Ultimately, 8 recommendations were formed by the GRADE grid method, while 16 consensus suggestions were reached via the majority vote rule. The results showed that the Binghuang Fule Ointment was applicable to eczema, psoriasis, neurodermatitis, tinea pedis, and other diseases. The Consensus also elucidated the syndrome differentiation points, usage and dosage for different diseases (such as adjustment of course and application frequency), as well as the indications of combination medication. Additionally, safety assessment suggested that the Ointment should be used with caution in individuals with skin ulceration or hypersensitivity. To ensure methodological rigor, the compilation process went through three rounds of internal and external expert reviews, while a comprehensive analysis was conducted by literature analysis, the Delphi method, and other methods. This compilation instruction provided methodological support for the clinical transformation of the Consensus through key links, including project initiation, international registration, informed consent, conflict-of-interest statements, evidence evaluation, and popularization. The Consensus will be continuously improved through a dynamic revision mechanism in the future.
5.Expert Consensus on Clinical Application of SkinPro Ointment
Xin CUI ; Ping SONG ; Yuanyuan LI ; Yanping BAI ; Yanming XIE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):163-170
SkinPro Ointment is an emulsion-based preparation derived from a traditional Tibetan medical empirical formula and developed using modern pharmaceutical technology. It is an exclusive patented product of Tibet Hairong Tangguo Pharmaceutical Co. Ltd. and has been listed as a National Protected Traditional Chinese Medicine Variety, the Pharmacopoeia of the People's Republic of China, and classified as a Category B product in the National Basic Medical Insurance Catalog. The ointment possesses the functions of clearing heat and drying dampness, activating blood circulation and dispelling wind, relieving itching and reducing inflammation. Clinically, it is used for skin pruritus caused by dampness-heat accumulation or blood-heat with wind-dryness, as well as pruritic skin diseases such as neurodermatitis, eczema, tinea pedis, and psoriasis. To clarify the standards for its clinical application and promote rational drug use, a consensus working group comprising 34 national experts in dermatology, evidence-based medicine, and pharmacy was established. Through expert interviews, the nominal group technique, and questionnaire surveys, 15 clinical issues were identified. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) evidence grading system was employed to assess the quality of evidence, leading to the formulation of the Expert Consensus on the Clinical Application of SkinPro Ointment. This consensus specifies that the intended users are physicians and pharmacists in medical institutions at all levels. It standardizes the clinical application of the ointment, including syndrome characteristics, dosage and course of treatment, combination therapy, precautions, and contraindications. Recommendations and consensus suggestions were formed addressing the 15 clinical issues, covering the following key areas: ①Indications and TCM syndromes: In TCM, the ointment mainly treats conditions such as "damp sores" (Shichuang), "white scaling" (Baibi), "collar sores" (Shelingchuang), and "damp foot Qi" (Jiaoshiqi), corresponding to eczema, psoriasis, neurodermatitis, and tinea pedis in Western medicine. The relevant TCM syndromes are identified as dampness-heat accumulation or blood-heat with wind-dryness. ②Usage and dosage: For external use, apply to the affected area 3 times daily. The dosage should follow the fingertip unit (FTU) principle. A treatment course of 1-2 weeks is recommended for mild to moderate cases; for serious cases, the course should be followed as prescribed by a physician. ③Combined therapy: The ointment can be used as monotherapy for mild cases. For moderate to severe cases, combination therapy is recommended, with reference to relevant clinical guidelines. ④Safety: Common adverse reactions include skin rashes, pruritus, and erythema. The ointment is contraindicated in patients with broken skin or obvious exudation at the affected area, as well as in patients with known hypersensitivity to any of its components. Contact with sensitive areas such as the eyes and oral mucosa should be avoided. Modern research shows that the ointment also has potential efficacy in other dermatological conditions, such as adult atopic dermatitis, tinea cruris, exfoliative keratolysis, acne vulgaris, and Malassezia folliculitis. This consensus provides a scientific basis for promoting the rational clinical use of SkinPro Ointment, improving its therapeutic efficacy, and reducing medication risks. Future updates will be dynamically revised according to emerging clinical issues and new evidence.
6.Effect of different exercise interventions on patients with metabolic dysfunction-associated fatty liver disease: A systematic review and network Meta-analysis
Guodong MA ; Zhuojing SUN ; Song HU ; Zijun YE ; Mingchen MA ; Fei CUI ; Jiaju ZHU
Journal of Clinical Hepatology 2026;42(2):326-344
ObjectiveTo investigate the effect of different exercise interventions on metabolism and liver parameters in patients with metabolic dysfunction-associated fatty liver disease (MAFLD), and to provide evidence-based recommendations for clinical exercise rehabilitation. MethodsThis study was conducted according to the PRISMA guidelines, and the protocol was registered on the PROSPERO platform, with a registration number of CRD42025641717. PubMed, Web of Science, Scopus, Wiley Online Library, CNKI, Wanfang Data, and VIP were searched for related articles published up to September 2024. The Cochrane tool for assessing risk of bias was used to assess the quality of articles, and Stata MP 17.0 was used to perform the network meta-analysis. ResultsA total of 57 articles were included, involving 2 648 patients. The results showed that aerobic exercise combined with resistance exercise had the best effect in improving body mass index (mean difference [WMD]=-0.97, 95% confidence interval [CI]: -1.66 to -0.28], P<0.05, surface under the cumulative ranking curve [SUCRA]=85.4) and triglycerides (WMD=-29.6, 95%CI: -46.66 to 12.54, P<0.05, SUCRA=87.3); resistance exercise was the optimal intervention method for improving total cholesterol (WMD=-15.99, 95%CI: -24.19 to -7.79, P<0.05, SUCRA=79.9) and glutamine transaminase (WMD=-8.08, 95%CI: -12.13 to -4.02, P<0.05, SUCRA=87.3); low-intensity aerobic exercise had the best effect in improving aspartate aminotransferase (WMD=-4.3, 95%CI: -8.45 to -0.15, P<0.05, SUCRA=73.5), gamma-glutamyl transpeptidase (GGT) (WMD=-3.26, 95%CI: -7.79 to 1.27, P>0.05, SUCRA=82.3), and glycated hemoglobin (HbA1c) (WMD=-0.6, 95%CI: -2.02 to 0.82, P>0.05, SUCRA=78.8); moderate-intensity aerobic exercise was the optimal intervention modality to improve Homeostasis Model Assessment of Insulin Resistance (WMD=-0.92, 95%CI: -1.51 to -0.33, P<0.05, SUCRA=69.4). It should be noted that there were no significant differences in HbA1c and GGT across different exercise interventions (all P>0.05), suggesting that there was currently no sufficient statistical evidence to support that exercise could improve these two indicators. ConclusionBased on the comprehensive league table and cumulative probability ranking, aerobic exercise combined with resistance exercise, resistance exercise, and low- and moderate-intensity aerobic exercise may be the best exercise modality for improving key indicators in MAFLD patients, and targeted exercise modalities should be selected for intervention against different indicators; however, due to limitations of the original studies, further studies are needed for validation and exploration.
7.Electroacupuncture Ameliorates NLRP3-mediated Pyroptosis in Spinal Cord Injury Rats by Reshaping The Gut Microbiota
Yin-Jie CUI ; Hong-Ru LI ; Jing-Yi LIU ; Hai-Lin DU ; Shu-Wen LIU ; Yuan YANG ; Chen-Guang ZHENG ; Jian-Qin XIANG ; Xiao-Juan SONG
Progress in Biochemistry and Biophysics 2026;53(5):1132-1153
ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.
8.Electroacupuncture Ameliorates NLRP3-mediated Pyroptosis in Spinal Cord Injury Rats by Reshaping The Gut Microbiota
Yin-Jie CUI ; Hong-Ru LI ; Jing-Yi LIU ; Hai-Lin DU ; Shu-Wen LIU ; Yuan YANG ; Chen-Guang ZHENG ; Jian-Qin XIANG ; Xiao-Juan SONG
Progress in Biochemistry and Biophysics 2026;53(5):1132-1153
ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.
9.Effect of Gypenosides on MAFLD Mice and Its Molecular Mechanism Based on Classical/Non-classical Ferroptosis Pathways
Yu LIU ; Yupeng PEI ; Jiaxin WANG ; Jingxuan ZHU ; Xiaofei SUN ; Qun WANG ; Peng CUI ; Nan SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):99-107
ObjectiveTo explore the effect of gypenosides (GPs) on liver lipid deposition in metabolism-associated fatty liver disease (MAFLD) mice and its mechanism based on classical/non-classical ferroptosis. MethodsEight male C57BL/6 mice in a blank group and 32 male apolipoprotein E gene knockout (ApoE-/-) mice were randomly divided into a model group, a low-dose GPs (GPs-L) group, a high-dose GPs (GPs-H) group, and a simvastatin (SV) group. Starting from the second week, mice in the blank group were given a maintenance diet, and the other four groups were fed a high-fat diet daily. After eight weeks of feeding, mice in the GPs-L and GPs-H groups were given GPs of 1.487 mg·kg-1·d-1 and 2.973 mg·kg-1·d-1, respectively, and mice in the SV group were given simvastatin of 2.275 mg·kg-1·d-1. Mice in the blank group and the model group were given saline of equal volume by gavage for four weeks. The content of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the serum of mice in each group was detected by an automatic biochemical analyzer. The level of non-esterified fatty acid (NEFA) and TG in the mouse liver was measured by the kit. The change in liver tissue structure and lipid deposition was observed by hematoxylin-eosin (HE) and oil red O staining. The levels of coenzyme Q10 (CoQ10), glutathione (GSH), malondialdehyde (MDA), and Fe2+ in serum, as well as nicotinamide adenine dinucleotide phosphate [NAD(P)H] in the liver were detected by enzyme-linked immunosorbent assay (ELISA). The expression of ferroptosis suppressor protein 1 (FSP1) in the liver of mice was observed by the immunohistochemical (IHC) method, and the expression of genes and proteins related to classical and non-classical ferroptosis pathways was analyzed by real-time polymerase chain reaction (Real-time PCR) and Wes automated protein expression analysis system. ResultsCompared with those in the blank group, the levels of TC, TG, LDL-C, ALT, and AST in serum and TG and NEFA in the liver in the model group were significantly increased, and the level of HDL-C in serum was significantly decreased (P<0.01). The liver tissue structure changed, and there were fat vacuoles of different sizes and a large number of red lipid droplets, with obvious lipid deposition. The level of CoQ10 and GSH in serum and NADH in the liver were significantly decreased, while the level of MDA and Fe2+ in serum was significantly increased (P<0.01). The mRNA and protein expressions of cystine/glutamate transporter (xCT/SLC7A11), glutathione peroxidase (GPX4), p62, nuclear factor E2-related factor 2 (Nrf2), and FSP1 were significantly decreased, and the mRNA and protein expressions of tumor antigen (p53), spermidine/spermine N1-acetyltransferase 1 (SAT1), arachidonate 15-lipoxygenase (ALOX15), and Kelch-like epichlorohydrin-associated protein-1 (Keap1) were significantly increased (P<0.01). Compared with those in the model group, the level of TC, TG, LDL-C, ALT, and AST in serum and TG and NEFA in the liver of mice in the GPs-L, GPs-H, and SV groups were decreased, while the level of HDL-C in serum was significantly increased (P<0.05, P<0.01). The liver tissue structure and lipid deposition were improved. The levels of CoQ10 and GSH in serum and NADH in the liver were significantly increased, while the levels of MDA and Fe2+ in serum were significantly decreased (P<0.05, P<0.01). The mRNA and protein expressions of xCT, GPX4, p62, Nrf2, and FSP1 were significantly increased, while the mRNA and protein expressions of p53, SAT1, ALOX15, and Keap1 were significantly decreased (P<0.05, P<0.01). ConclusionGPs can interfere with liver lipid deposition in MAFLD mice through classical/non-classical ferroptosis pathways.
10.Effect of Gypenosides on MAFLD Mice and Its Molecular Mechanism Based on Classical/Non-classical Ferroptosis Pathways
Yu LIU ; Yupeng PEI ; Jiaxin WANG ; Jingxuan ZHU ; Xiaofei SUN ; Qun WANG ; Peng CUI ; Nan SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):99-107
ObjectiveTo explore the effect of gypenosides (GPs) on liver lipid deposition in metabolism-associated fatty liver disease (MAFLD) mice and its mechanism based on classical/non-classical ferroptosis. MethodsEight male C57BL/6 mice in a blank group and 32 male apolipoprotein E gene knockout (ApoE-/-) mice were randomly divided into a model group, a low-dose GPs (GPs-L) group, a high-dose GPs (GPs-H) group, and a simvastatin (SV) group. Starting from the second week, mice in the blank group were given a maintenance diet, and the other four groups were fed a high-fat diet daily. After eight weeks of feeding, mice in the GPs-L and GPs-H groups were given GPs of 1.487 mg·kg-1·d-1 and 2.973 mg·kg-1·d-1, respectively, and mice in the SV group were given simvastatin of 2.275 mg·kg-1·d-1. Mice in the blank group and the model group were given saline of equal volume by gavage for four weeks. The content of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the serum of mice in each group was detected by an automatic biochemical analyzer. The level of non-esterified fatty acid (NEFA) and TG in the mouse liver was measured by the kit. The change in liver tissue structure and lipid deposition was observed by hematoxylin-eosin (HE) and oil red O staining. The levels of coenzyme Q10 (CoQ10), glutathione (GSH), malondialdehyde (MDA), and Fe2+ in serum, as well as nicotinamide adenine dinucleotide phosphate [NAD(P)H] in the liver were detected by enzyme-linked immunosorbent assay (ELISA). The expression of ferroptosis suppressor protein 1 (FSP1) in the liver of mice was observed by the immunohistochemical (IHC) method, and the expression of genes and proteins related to classical and non-classical ferroptosis pathways was analyzed by real-time polymerase chain reaction (Real-time PCR) and Wes automated protein expression analysis system. ResultsCompared with those in the blank group, the levels of TC, TG, LDL-C, ALT, and AST in serum and TG and NEFA in the liver in the model group were significantly increased, and the level of HDL-C in serum was significantly decreased (P<0.01). The liver tissue structure changed, and there were fat vacuoles of different sizes and a large number of red lipid droplets, with obvious lipid deposition. The level of CoQ10 and GSH in serum and NADH in the liver were significantly decreased, while the level of MDA and Fe2+ in serum was significantly increased (P<0.01). The mRNA and protein expressions of cystine/glutamate transporter (xCT/SLC7A11), glutathione peroxidase (GPX4), p62, nuclear factor E2-related factor 2 (Nrf2), and FSP1 were significantly decreased, and the mRNA and protein expressions of tumor antigen (p53), spermidine/spermine N1-acetyltransferase 1 (SAT1), arachidonate 15-lipoxygenase (ALOX15), and Kelch-like epichlorohydrin-associated protein-1 (Keap1) were significantly increased (P<0.01). Compared with those in the model group, the level of TC, TG, LDL-C, ALT, and AST in serum and TG and NEFA in the liver of mice in the GPs-L, GPs-H, and SV groups were decreased, while the level of HDL-C in serum was significantly increased (P<0.05, P<0.01). The liver tissue structure and lipid deposition were improved. The levels of CoQ10 and GSH in serum and NADH in the liver were significantly increased, while the levels of MDA and Fe2+ in serum were significantly decreased (P<0.05, P<0.01). The mRNA and protein expressions of xCT, GPX4, p62, Nrf2, and FSP1 were significantly increased, while the mRNA and protein expressions of p53, SAT1, ALOX15, and Keap1 were significantly decreased (P<0.05, P<0.01). ConclusionGPs can interfere with liver lipid deposition in MAFLD mice through classical/non-classical ferroptosis pathways.

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