1.Neuroprotective Mechanism of Yifei Xuanfei Jiangzhuo Prescription on VaD Rats Based on NF-κB/NLRP3 Signaling Pathway
Bingmao YUAN ; Wei CHEN ; Xiu LAN ; Lingfei JIANG ; Lin WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):88-96
ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway to improve neuronal function in vascular dementia (VaD) rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries (CCA) combined with bilateral vascular occlusion (2-VO). Eighty-four SD rats were randomly divided into a blank group, sham group, model group, piracetam group (0.2 g·kg-1), and low-, medium-, and high-dose Yifei Xuanfei Jiangzhuo prescription groups (6.09, 12.18, and 24.36 g·kg-1). Drug administration began on day 7 after surgery, once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin (HE) staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen (NeuN). Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase (IKK), NF-κB p65, NLRP3, Caspase-1, apoptosis-associated speck-like protein (ASC), and interleukin-1β (IL-1β). ResultsCompared with the blank group, the model group showed a significant reduction in platform-crossing frequency (P0.01), aggravated hippocampal injury, a significant increase in neuronal apoptosis (P0.05), decreased NeuN positivity in the CA1 region (P0.05), increased nuclear expression of NF-κB p65 (P0.05), and significantly elevated expression of p-IKK, p-NF-κB p65, NLRP3, cleaved Caspase-1, ASC, and cleaved IL-1β (P0.05). Compared with the model group, all drug-treated groups improved learning and spatial memory in VaD rats, alleviated hippocampal pathological injury and neuronal apoptosis, and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg-1 reduced hippocampal expression levels of p-IKK, p-NF-κB p65, NLRP3, Caspase-1, ASC, and cleaved IL-1β in VaD rats (P0.05), showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway, thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.
2.Clinical Efficacy of Qi-regulating and Phlegm-removing Method(Liu Junzitang Combined with Linggang Wuwei Jiangxintang) in Treating AECOPD with Increased EOS
Renjie HUANG ; Wangqin YU ; Wuyinuo TANG ; Hong SONG ; Lyuyuan HE ; Wenbo LIN ; Guanyi WU ; Hang HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):149-156
ObjectiveTo assess the efficacy and safety of the Qi-regulating and phlegm-removing method(Liu Junzitang Combined with Linggang Wuwei Jiangxintang) for treating acute exacerbations of chronic obstructive pulmonary disease (AECOPD) with increased eosinophils (EOS). MethodsSixty-eight AECOPD patients with increased EOS who were hospitalized in the Department of Pulmonary Diseases of Jinhua Traditional Chinese Medicine Hospital from April 2023 to April 2024 were recruited and randomly assigned to an experimental group (EG) or a control group (CG). Both groups received conventional Western medicine, with the EG additionally receiving Liujunzitang and Linggan Wuwei Jiangxintang. The therapeutic efficacy indicators were measured after the treatment. The main therapeutic efficacy indicators included partial pressure of oxygen (PaO2) and partial pressure of carbon dioxide (PaCO2). The secondary efficacy indicators included the TCM symptom scores, the COPD Assessment Test (CAT) score, the Modified Medical Research Council (mMRC) Dyspnea Scale score, and the length of hospital stay. The indicators were measured at baseline and on days 3 and 7 of intervention. The safety was evaluated based on the adverse events. ResultsBaseline characteristics were not statistically different between the two groups. Compared with CG, EG showed no significant difference in PaO2 (P=0.773), PaCO2 (P=0.632) and or CAT score (P=0.336) at on day 3 but better PaO2 (P=0.004), PaCO2 (P=0.008), and CAT score (P=0.013) were significantly better at on day 7. Compared with CGAfter treatment, EG had lower TCM syndrome scores of than CG EG on day 3 (P=0.005) and day 7 were significantly decreased (P0.001). There was no significant difference in mMRC score between the two groups on day 3 (P=0.514) and day 7 (P=0.176) as wasor the length of hospital stay (P=0.915). The generalized linear mixed model (GLMM) showed that compared with CG, EG had significant improvements over time in PaO2, PaCO2, TCM syndrome symptom scores, CAT score, and mMRC score. ConclusionRegulating qi Qi and removing phlegm combined with conventional Western medicine can significantly alleviateimprove the clinical symptoms and improve the lung function of AECOPD patients with increased EOS increased AECOPDwhich has and demonstrates good safety.
3.Neuroprotective Mechanism of Yifei Xuanfei Jiangzhuo Prescription on VaD Rats Based on NF-κB/NLRP3 Signaling Pathway
Bingmao YUAN ; Wei CHEN ; Xiu LAN ; Lingfei JIANG ; Lin WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):88-96
ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway to improve neuronal function in vascular dementia (VaD) rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries (CCA) combined with bilateral vascular occlusion (2-VO). Eighty-four SD rats were randomly divided into a blank group, sham group, model group, piracetam group (0.2 g·kg-1), and low-, medium-, and high-dose Yifei Xuanfei Jiangzhuo prescription groups (6.09, 12.18, and 24.36 g·kg-1). Drug administration began on day 7 after surgery, once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin (HE) staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen (NeuN). Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase (IKK), NF-κB p65, NLRP3, Caspase-1, apoptosis-associated speck-like protein (ASC), and interleukin-1β (IL-1β). ResultsCompared with the blank group, the model group showed a significant reduction in platform-crossing frequency (P0.01), aggravated hippocampal injury, a significant increase in neuronal apoptosis (P0.05), decreased NeuN positivity in the CA1 region (P0.05), increased nuclear expression of NF-κB p65 (P0.05), and significantly elevated expression of p-IKK, p-NF-κB p65, NLRP3, cleaved Caspase-1, ASC, and cleaved IL-1β (P0.05). Compared with the model group, all drug-treated groups improved learning and spatial memory in VaD rats, alleviated hippocampal pathological injury and neuronal apoptosis, and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg-1 reduced hippocampal expression levels of p-IKK, p-NF-κB p65, NLRP3, Caspase-1, ASC, and cleaved IL-1β in VaD rats (P0.05), showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway, thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.
4.Clinical Efficacy of Qi-regulating and Phlegm-removing Method(Liu Junzitang Combined with Linggang Wuwei Jiangxintang) in Treating AECOPD with Increased EOS
Renjie HUANG ; Wangqin YU ; Wuyinuo TANG ; Hong SONG ; Lyuyuan HE ; Wenbo LIN ; Guanyi WU ; Hang HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):149-156
ObjectiveTo assess the efficacy and safety of the Qi-regulating and phlegm-removing method(Liu Junzitang Combined with Linggang Wuwei Jiangxintang) for treating acute exacerbations of chronic obstructive pulmonary disease (AECOPD) with increased eosinophils (EOS). MethodsSixty-eight AECOPD patients with increased EOS who were hospitalized in the Department of Pulmonary Diseases of Jinhua Traditional Chinese Medicine Hospital from April 2023 to April 2024 were recruited and randomly assigned to an experimental group (EG) or a control group (CG). Both groups received conventional Western medicine, with the EG additionally receiving Liujunzitang and Linggan Wuwei Jiangxintang. The therapeutic efficacy indicators were measured after the treatment. The main therapeutic efficacy indicators included partial pressure of oxygen (PaO2) and partial pressure of carbon dioxide (PaCO2). The secondary efficacy indicators included the TCM symptom scores, the COPD Assessment Test (CAT) score, the Modified Medical Research Council (mMRC) Dyspnea Scale score, and the length of hospital stay. The indicators were measured at baseline and on days 3 and 7 of intervention. The safety was evaluated based on the adverse events. ResultsBaseline characteristics were not statistically different between the two groups. Compared with CG, EG showed no significant difference in PaO2 (P=0.773), PaCO2 (P=0.632) and or CAT score (P=0.336) at on day 3 but better PaO2 (P=0.004), PaCO2 (P=0.008), and CAT score (P=0.013) were significantly better at on day 7. Compared with CGAfter treatment, EG had lower TCM syndrome scores of than CG EG on day 3 (P=0.005) and day 7 were significantly decreased (P0.001). There was no significant difference in mMRC score between the two groups on day 3 (P=0.514) and day 7 (P=0.176) as wasor the length of hospital stay (P=0.915). The generalized linear mixed model (GLMM) showed that compared with CG, EG had significant improvements over time in PaO2, PaCO2, TCM syndrome symptom scores, CAT score, and mMRC score. ConclusionRegulating qi Qi and removing phlegm combined with conventional Western medicine can significantly alleviateimprove the clinical symptoms and improve the lung function of AECOPD patients with increased EOS increased AECOPDwhich has and demonstrates good safety.
5.Mechanism of inhibitory effect of total flavonoids from Taraxacum mongolicum on obesity in mice by regulating intestinal flora
Yixue GAO ; Lin GUO ; Linyan LANG ; Jing WU ; Haoyang WANG ; Jing YANG ; Mingsan MIAO ; Zhanzhan LI
China Pharmacy 2025;36(3):293-299
OBJECTIVE To investigate the mechanism of the inhibitory effect of total flavonoids from Taraxacum mongolicum on high-fat diet-induced obesity in mice through modulation of intestinal flora. METHODS Twenty-four C57BL/6J mice were randomly divided into blank group, model group and T. mongolicum total flavonoid group, with 8 mice in each group. Except for the blank group, the other 2 groups were given a high-fat diet, while T. mongolicum total flavonoid group was given T. mongolicum total flavonoid [400 mg/(kg·d)] intragastrically, once a day, for 8 consecutive weeks. During the experiment, the food intake of each group of mice was recorded. After the last medication, the body mass, fat weight, blood lipid level and pathological changes of liver and epididymal fat in mice were evaluated to observe the effect of T. mongolicum total flavonoid on the treatment of obesity in mice. The changes in abundance and structure of intestinal flora in mice were detected by amplicon sequencing; the effects of T. mongolicum total flavonoids on fat metabolism related genes were analyzed by qPCR. RESULTS Compared with model group, the body weight of mice in T. mongolicum total flavonoids group was decreased significantly (P<0.05); the levels of total lipid cholesterol, triglycerides, and LDL cholesterol were all decreased significantly (P<0.01), and the level of HDL cholesterol was increased significantly (P<0.01); the fat indexes of inguinal white adipose tissue and epididymal white wind_lz@hactcm.edu.cn adipose tissue were significantly reduced (P<0.05); significant improvement in hepatocellular steatosis and adipose cytopathy were significantly improved; mRNA expressions of COX7A1 and COX8B were significantly upregulated (P<0.05). The results of bacterial colony detection showed that compared with the model group, there was a rising trend in the diversity of the bacterial colony in T. mongolicum total flavonoids group, and the Sobs index characterization and β diversity were increased significantly (P<0.05). Relative abundances of Blautia, norank_f_Ruminococcaceae, Bilophila, Alistipes, classified_f_Ruminococcaceae, Parabacteroides, norank_f_Desulfovibrionaceae, Anaerotruncus were significantly up-regulated(P<0.05), while those of Faecalibaculum, Erysipelatoclostridium, GCA-900066575, Tuzzerella, Lactobacillus, norank_f_norank_o_RF39, achnospiraceae_FCS020_group were significantly down-regulated (P<0.05). CONCLUSIONS T. mongolicum total flavonoids can reduce body mass, fat weight and blood lipid levels, and repair the pathological damage to liver and epididymal fat in obese mice, which is related to improving intestinal flora disorders caused by high-fat diet.
6.Working practices in eliminating the public health crisis caused by viral hepatitis in Hainan Province of China
Weihua LI ; Changfu XIONG ; Taifan CHEN ; Bin HE ; Dapeng YIN ; Xuexia ZENG ; Feng LIN ; Biyu CHEN ; Xiaomei ZENG ; Biao WU ; Juan JIANG ; Lu ZHONG ; Yuhui ZHANG
Journal of Clinical Hepatology 2025;41(2):228-233
In 2022, Hainan provincial government launched the project for the prevention and control of viral hepatitis with the goals of a hepatitis B screening rate of 90%, a diagnostic rate of 90%, and a treatment rate of 80% among people aged 18 years and above by the year 2025, and the main intervention measures include population-based prevention, case screening, antiviral therapy, and health management. As of December 31, 2024, a total of 6.875 million individuals in the general population had been screened for hepatitis B, with a screening rate of 95.6%. A total of 184 710 individuals with positive HBsAg were identified, among whom 156 772 were diagnosed through serological reexamination, resulting in a diagnostic rate of 84.9%. A total of 50 742 patients with chronic hepatitis B were identified, among whom 42 921 had hepatitis B-specific health records established for health management, with a file establishment rate of 84.6%. A total of 31 553 individuals received antiviral therapy, with a treatment rate of 62.2%. A total of 2.503 million individuals at a high risk of hepatitis C were screened, among whom 4 870 tested positive for HCV antibody and 3 858 underwent HCV RNA testing, resulting in a diagnostic rate of 79.2%, and 1 824 individuals with positive HCV RNA were identified, among whom 1 194 received antiviral therapy, with a treatment rate of 65.5%. In addition, 159 301 individuals with negative HBsAg and anti-HBs and an age of 20 — 40 years were inoculated with hepatitis B vaccine free of charge. Through the implementation of the project for the prevention and control of viral hepatitis, a large number of hepatitis patients have been identified, treated, and managed in the province within a short period of time, which significantly accelerates the efforts to eliminate the crisis of viral hepatitis.
7.Fitting Degrees of Animal Models of Chronic Glomerulonephritis with Clinical Characteristics in Western Medicine and Traditional Chinese Medicine
Siqing WU ; Lin GUO ; Beibei ZHANG ; Mingsan MIAO ; Jinxin MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):259-264
Chronic glomerulonephritis (CGN) is a common clinical chronic glomerular disease caused by autoimmune reaction, the pathogenesis of which is complex and has not been fully elucidated. There is no specific treatment method in modern medicine. The establishment of an animal model of CGN in accordance with its characteristics in western medicine and traditional Chinese medicine will help to reveal the pathogenesis of CGN, rate drugs, and improve the treatment plan. Based on the clinical diagnostic criteria of CGN, the paper establishes the syndrome differentiation criteria of CGN for Chinese and western medicine. Through summarizing the literature on animal models of CGN and making a further analysis, it is found that the CGN models are mainly modeled using rats with the methods of single-factor induction or two-factor induction, and the main manifestation of the disease characteristics is nephritis-related symptoms. The single-factor-induced or two-factor-induced CGN rat models have a high fitting degree with the clinical characteristics in western medicine, but the fitting degree is insufficient with the clinical characteristics in traditional Chinese medicine. In addition, the CGN models with syndromes of traditional Chinese medicine are dominated by Qi deficiency in the spleen and kidney and Qi deficiency in the lung and kidney, while models for Yang deficiency in the spleen and kidney, Yin deficiency in the liver and kidney, and deficiency of both Qi and Yin are slightly insufficient. Therefore, it is important to prepare a new and improved animal model of CGN, so that a preclinical model can be provided for the exploration of the pathogenesis of CGN in western medicine and traditional Chinese medicine and its therapeutic research.
8.Mechanism of Modified Shaofu Zhuyutang in Antagonising Ectopic Endometrial Tissue Fibrosis Based on Cellular Pyroptosis Mediated by TRL4/NF-κB/NLPR3 Signaling Pathway
Zuoliang ZHANG ; Jiaxing WANG ; Wanrun WANG ; Xiangyu LIN ; Bin YUE ; Zhirui ZHANG ; Yinan WANG ; Yaling YANG ; Dongqing WEI ; Cancan HUANG ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):19-28
ObjectiveTo investigate the mechanism of action of modified Shaofu Zhuyutang in antagonizing cellular pyroptosis and fibrosis in ectopic endometrial tissues of endometriosis through the Toll-like receptor 4/nuclear factor-κB/NOD-like receptor protein 3 (TRL4/NF-κB/NLPR3) signaling pathway. MethodsSeventy-two SPF-grade female SD rats were randomly divided into a sham-operated group (n = 12) and a modeling group (n = 60). The rats in the sham-operated group underwent a caesarean section, while the rats in the modeling group were used to establish an endometriosis model through the auto-transplantation method. After successful modeling, the animals were randomly divided into the model group, progesterone group (0.25 mg·kg-1), and modified Shaofu Zhuyutang low-, medium-, and high-dose groups (7.5, 15, 30 g·kg-1), with 12 animals in each group. After 4 weeks of drug administration, voluntary activity and heat pain latency were observed. The rats were sacrificed for tissue collection, and Masson staining were used to observe histopathological changes in the endometrial tissues. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of interleukin-18 (IL-18), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and transforming growth factor-β (TGF-β). Immunohistochemistry (IHC) was used to detect the protein expression area of tumor necrosis factor-related factor 6 (TRAF6) and NLPR3 in the endometrial tissues. Immunofluorescence (IF) was used to detect the relative fluorescence intensity of Caspase-1 and gasdermin D (GSDMD) in the endometrial tissues. Western blot was employed to measure the relative expression of TRL4, myeloid differentiation factor 88 (MyD88), TRAF6, NF-κB p65, phosphorylated NF-κB p65 (p-NF-κB p65), and NLPR3 proteins in endometrial tissues. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of TRL4, MyD88, TRAF6, NF-κB, and NLPR3 in the endometrial tissues. ResultsCompared with the sham-operated group, rats in the model group showed reduced voluntary activity and shorter heat pain latency. Serum levels of IL-18, IL-1β, TNF-α, and TGF-β were elevated. The relative expression areas of TRAF6 and NLPR3 proteins were increased, and the relative fluorescence intensity of Caspase-1 and GSDMD was enhanced. The relative expression of TRL4, MyD88, TRAF6, NF-κB p65, p-NF-κB p65, and NLPR3 proteins, along with the expression of TRL4, MyD88, TRAF6, NF-κB, and NLPR3 mRNA, were significantly increased (P<0.01). Compared with the model group, rats in the progesterone group and the modified Shaofu Zhuyutang medium- and high-dose groups exhibited improved voluntary activity, longer heat pain latency, the fibrosis of endometrial tissue is alleviated. Serum levels of IL-18, IL-1β, TNF-α, and TGF-β were decreased. The relative expression areas of TRAF6 and NLPR3 proteins decreased, and the relative fluorescence intensity of Caspase-1 and GSDMD weakened. The relative expression of TRL4, MyD88, TRAF6, p-NF-κB p65, NLPR3 proteins, and TRL4, MyD88, TRAF6, NF-κB, and NLPR3 mRNA expression were reduced (P<0.05, P<0.01). ConclusionModified Shaofu Zhuyutang may play a therapeutic role in endometriosis by interfering with key proteins in the TRL4/NF-κB/NLPR3 signaling pathway, reducing NLRP3 inflammasome-induced cellular pyroptosis, antagonizing the fibrosis process in ectopic endometrial tissues, improving the inflammatory microenvironment in the pelvic cavity, and alleviating pain.
9.Mechanism of Modified Shaofu Zhuyutang in Treatment of Endometriosis Based on EGFR/PI3K/Akt Signaling Pathway
Yaling YANG ; Wanrun WANG ; Zuoliang ZHANG ; Xiangyu LIN ; Jiaxing WANG ; Cancan HUANG ; Xiujia JI ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):29-38
ObjectiveTo observe the effects of modified Shaofu Zhuyutang on key proteins of the epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in SD rats with endometriosis. MethodsAfter successful establishment of an endometriosis model in 60 female SD rats of SPF grade via the auto-transplantation method, the rats were randomly divided into a model group, modified Shaofu Zhuyutang high-, medium-, and low-dose groups, and a gestrinone group, with another 12 rats serving as a blank group. The blank and model groups were administered 10 mL·kg-1 normal saline, while the high-, medium-, and low-dose groups received 30, 15, and 7.5 g·kg-1 modified Shaofu Zhuyutang, respectively. The gestrinone group was administered 0.25 mg·kg-1 gestrinone suspension. After four weeks of treatment, uterine contractions were induced with 2 U of oxytocin, and the writhing response of rats was observed. After 24 h, the rats were euthanized, and the weight and volume of ectopic endometrial tissue were recorded. Hematoxylin-eosin (HE) staining was used to observe pathological changes in endometrial tissues, while the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay was used to evaluate the apoptosis rate of endometrial tissues. Immunofluorescence was used to detect the relative expression areas of the B-cell lymphoma-2 gene-associated promoter (Bad) and B-cell lymphoma-2 (Bcl-2) proteins in endometrial tissues. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), epidermal growth factor (EGF), and EGFR were measured by enzyme-linked immunosorbent assay (ELISA). The relative protein expression levels of EGFR, PI3K, phosphorylated PI3K (p-PI3K), Akt, and phosphorylated Akt (p-Akt) in endometrial tissues were analyzed by Western blot. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of EGFR, PI3K, and Akt. ResultsCompared with the blank group, the model group showed endometrial thickening, glandular and mesenchymal hyperplasia, a significant decrease in the relative expression area of Bad in ectopic endometrial tissues, a significant increase in the relative expression area of Bcl-2, and a significant reduction in the apoptosis rate as indicated by TUNEL staining. Serum levels of IL-1β, IL-6, TNF-α, EGF, and EGFR were significantly elevated (P<0.01). The relative protein expression levels of EGFR, PI3K, p-PI3K, Akt, and p-Akt, as well as the mRNA expression levels of EGFR, PI3K, and Akt, were also significantly increased (P<0.01). Compared with the model group, the high- and medium-dose groups of modified Shaofu Zhuyutang and the gestrinone group exhibited reduced glandular and mesenchymal hyperplasia to varying degrees, with dilated glandular lumens. The number of writhing responses was significantly reduced, the latency to writhing response was significantly prolonged, and the weight and volume of ectopic endometrial tissue were significantly decreased. The relative expression area of Bad in ectopic endometrial tissue was significantly increased, the relative expression area of Bcl-2 was significantly decreased, and the apoptosis rate was significantly elevated as shown by TUNEL staining. Serum levels of IL-1β, IL-6, TNF-α, EGF, and EGFR were significantly reduced, and the relative protein expression levels of EGFR, PI3K, p-PI3K, Akt, and p-Akt, as well as the mRNA expression levels of EGFR, PI3K, and Akt, were significantly decreased (P<0.05,P<0.01). ConclusionModified Shaofu Zhuyutang may exert therapeutic effects on endometriosis by interfering with key proteins of the EGFR/PI3K/Akt signaling pathway and inducing apoptosis in ectopic endometrial tissue.
10.Effect of Modified Shaofu Zhuyutang on Ferroptosis in Ectopic Endometrial Tissues of Rats with Endometriosis Based on MDM4/p53/GPX4 Signaling Pathway
Zuoliang ZHANG ; Xiangyu LIN ; Wanrun WANG ; Jiaxing WANG ; Yaling YANG ; Quansheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):39-47
ObjectiveTo investigate the mechanism of modified Shaofu Zhuyutang in inducing ferroptosis in ectopic endometrial tissues of rats with endometriosis through the murine double minute 4 (MDM4)/tumor suppressor p53/glutathione peroxidase 4 (GPX4) signaling pathway. MethodsSeventy SPF-grade female SD rats were randomly divided into a blank group (n = 10), a sham-operated group (n = 10), and a modeling group (n = 50). The sham-operated group underwent laparotomy, while the modeling group was subjected to the autotransplantation method to establish an endometriosis model. After successful modeling, the animals were randomly assigned to the model group, progesterone group (0.25 mg·kg-1), and modified Shaofu Zhuyutang high-, medium-, and low-dose groups (30, 15, and 7.5 g·kg-1, respectively), with 10 rats per group. After four weeks of drug administration, the rats were euthanized for sample collection. The weight and volume of ectopic endometrial tissues were recorded for each group. Transmission electron microscopy (TEM) was employed to observe ultrastructural changes in endometrial tissues, while Prussian blue staining was used to assess iron ion deposition. Serum levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). The relative levels of Fe2+, malondialdehyde (MDA), and glutathione (GSH) in endometrial tissues were determined by colorimetric assay. Immunofluorescence (IF) was used to detect the relative fluorescence intensities of GSH and GPX4 in endometrial tissues. The relative expression levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (Akt), phosphorylated Akt (p-Akt), MDM4, p53, and GPX4 proteins were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to assess the mRNA expression of PI3K, Akt, MDM4, p53, and GPX4. ResultsCompared with the blank and sham-operated groups, the model group exhibited reduced ferroptotic damage in ultrastructural observations, decreased ferroptotic aggregates and positive iron ion expression area on Prussian blue staining, elevated serum IL-6, IL-1β, and TNF-α levels, reduced Fe2+ and MDA content, increased GSH content in endometrial tissues, and enhanced GSH and GPX4 fluorescence intensities (P<0.01). The protein and mRNA expression levels of PI3K, p-PI3K, Akt, p-Akt, MDM4, and GPX4 were elevated, while those of p53 were decreased (P<0.01). Compared with the model group, in the progesterone group and the modified Shaofu Zhuyutang high- and medium-dose groups, ferroptotic damage in ultrastructural observations was exacerbated to varying degrees by TEM, and ferroptotic aggregates and positive iron ion expression areas were increased on Prussian blue staining. Serum IL-6, IL-1β, and TNF-α levels decreased, Fe2+ and MDA content increased, and GSH content decreased in endometrial tissues. GSH and GPX4 fluorescence intensities weakened, while the protein and mRNA expression levels of PI3K, p-PI3K, Akt, p-Akt, MDM4, and GPX4 decreased, and those of p53 increased (P<0.05, P<0.01). Conclusionmodified Shaofu Zhuyutang may exert therapeutic effects in endometriosis by inducing ferroptosis in ectopic endometrial tissues, alleviating inflammatory responses, and modulating key proteins in the MDM4/p53/GPX4 signaling pathway.

Result Analysis
Print
Save
E-mail