1.Shentong Zhuyutang Regulates SIRT1/Nrf2 Pathway to Ameliorate Intervertebral Disc Degeneration in Rats
Jiajun HUANG ; Diyou WU ; Guangyi TAO ; Yu ZHAO ; Junqing HUANG ; Bin YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):29-39
ObjectiveTo study the effect and mechanism of Shentong Zhuyutang in treating intervertebral disc degeneration (IDD) in rats. MethodsIn the cell experiment, male rats were administrated with normal saline or low-, medium-, and high-dose (3.38, 6.75,13.5 g·kg-1, respectively) Shentong Zhuyutang by gavage, respectively, and serum samples were collected after 7 days of continuous administration. Another 10 male rats were selected for the isolation of nucleus pulposus cells. The cell model of IDD was established by treatment with interleukin (IL)-1β. The modeled cells were then treated with Shentong Zhuyutang-containing serum and the ferroptosis inhibitor ferrostatin-1 (Fer-1), respectively, to investigate the effects of Shentong Zhuyutang-containing serum on the proliferation and ferroptosis of nucleus pulposus cells. To study the role of silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) in the regulation of ferroptosis in nucleus pulposus cells by Shentong Zhuyutang-containing serum, this study treated the cells with the SIRT1 inhibitor Ex 527 and the Nrf2 inhibitor ML385, respectively, in addition to the treatment with IL-1β and high-dose Shentong Zhuyutang-containing serum. The cell-counting kit-8 (CCK-8) assay and EdU staining were employed to measure the cell viability and proliferation, respectively. The Fe2+, glutathione (GSH), and malondiadehyde (MDA) levels were measured by colorimetric assay. Western blot was employed to determine the protein levels of glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family 4 (ACSL4), Collagen Ⅱ, Aggrecan, SIRT1, and Nrf2. Immunofluorescence was used detect SIRT1 expression. In the animal experiment, male rats were treated with anulus puncture for the modeling of IDD. Rats were randomly assigned into sham operation, model, Shentong Zhuyutang-containing serum (13.5 g·kg-1), and positive control (nimesulide dispersible tablets, 0.18 mg·kg-1) groups. Rats in the drug intervention groups were administrated with corresponding agents at 1 mL·kg-1, and those in the sham operation and model groups were administrated with equal volumes of normal saline, once daily for 28 consecutive days. At the end of the last administration, the histopathological changes in the intervertebral discs of rats were observed by hematoxylin-eosin staining and scored by the Masuda method. Western blot was employed to determine the protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ in the nucleus pulposus tissue. ResultsCompared with the control group, the IL-1β group of nucleus pulposus cells showed elevated levels of Fe2+, MDA, and ACSL4 (P<0.05), decreased cell viability, lowered GSH level, and down-regulated protein levels of GPX4, Collagen Ⅱ, and Aggrecan (P<0.05). Shentong Zhuyutang-containing serum and Fer-1 reversed the effects of IL-1β on the viability and ferroptosis of nucleus pulposus cells and up-regulated the protein levels of Collagen Ⅱ and Aggrecan in nucleus pulposus cells (P<0.05). Compared with the control group, the IL-1β group showcased down-regulated expression of Sirt1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the IL-1β group, the high-dose Shentong Zhuyutang-containing serum+IL-1β group showed up-regulated expression of SIRT1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the high-dose Shentong Zhuyutang-containing serum+IL-1β group, the ML385 group showed down-regulated protein levels of Nrf2 and GPX4, lowered GSH level, and elevated Fe2+ and MDA levels (P<0.05). In addition, the Ex 527 group showed down-regulated protein levels of SIRT1, Nrf2, and GPX4 (P<0.05). The results of the animal experiment showed that compared with the sham operation group, the model group had severe degeneration of the intervertebral disc tissue with increased pathological score, up-regulated protein level of ACSL4 (P<0.05), and down-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). Compared with the model group, the Shentong Zhuyutang group showed alleviated IDD with declined pathological score, down-regulated protein level of ACSL4 (P<0.05), and up-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). ConclusionShentong Zhuyutang may activate the SIRT1/Nrf2 signaling pathway to inhibit the ferroptosis of nucleus pulposus cells, thereby delaying the process of IDD in rats.
2.Proteomics and Network Pharmacology Reveal Mechanism of Xiaoer Huatan Zhike Granules in Treating Allergic Cough
Youqi DU ; Yini XU ; Jiajia LIAO ; Chaowen LONG ; Shidie TAI ; Youwen DU ; Song LI ; Shiquan GAN ; Xiangchun SHEN ; Ling TAO ; Shuying YANG ; Lingyun FU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):69-79
ObjectiveTo explore the pharmacological mechanism involved in the treatment of allergic cough (AC) by Xiaoer Huatan Zhike granules (XEHT) based on proteomics and network pharmacology. MethodsAfter sensitization by intraperitoneal injection of 1 mL suspension containing 2 mg ovalbumin (OVA) and 100 mg aluminum hydroxide, a guinea pig model of allergic cough was constructed by nebulization with 1% OVA. The modeled guinea pigs were randomized into the model, low-, medium- and high-dose (1, 5, 20 g·kg-1, respectively) XEHT, and sodium montelukast (1 mg·kg-1) groups (n=6), and another 6 guinea pigs were selected as the blank group. The guinea pigs in drug administration groups were administrated with the corresponding drugs by gavage, and those in the blank and model groups received the same volume of normal saline by gavage, 1 time·d-1. After 10 consecutive days of drug administration, the guinea pigs were stimulated by 1% OVA nebulization, and the coughs were observed. The pathological changes in the lung tissue were observed by hematoxylin-eosin staining. The enzyme-linked immunosorbent assay was performed to measure the levels of C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), and malondialdehyde (MDA) in the bronchoalveolar lavage fluid (BALF) and immunoglobulin G (IgG) and immunoglobulin A (IgA) in the serum. Immunohistochemistry (IHC) was employed to observe the expression of IL-6 and TNF-α in the lung tissue. Transmission electron microscopy was employed observe the alveolar type Ⅱ epithelial cell ultrastructure. Real-time PCR was employed to determine the mRNA levels of IL-6, interleukin-1β (IL-1β), and TNF-α in the lung tissue. Label-free proteomics was used to detect the differential proteins among groups. Network pharmacology was used to predict the targets of XEHT in treating AC. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to search for the same pathways from the results of proteomics and network pharmacology. ResultsCompared with the blank group, the model group showed increased coughs (P<0.01), elevated levels of CRP, TNF-α, IL-6, and MDA and lowered level of SOD in the BALF (P<0.05, P<0.01), elevated levels of IgA and IgG in the serum (P<0.05, P<0.01), congestion of the lung tissue and infiltration of inflammatory cells, increased expression of IL-6 and TNF-α (P<0.01), large areas of low electron density edema in type Ⅱ epithelial cells, obvious swelling and vacuolization of the organelles, karyopyknosis or sparse and dissolved chromatin, and up-regulated mRNA levels of IL-6, IL-1β, and TNF-α (P<0.01). Compared with the model group, the drug administration groups showed reduced coughs (P<0.01), lowered levels of CRP, TNF-α, IL-6, and MDA and elevated level of SOD in the BALF (P<0.05, P<0.01), alleviated lung tissue congestion, inflammatory cell infiltration, and type Ⅱ epithelial cell injury, and decreased expression of IL-6 and TNF-α (P<0.01). In addition, the medium-dose XEHT group and the montelukast sodium group showcased lowered serum levels of IgA and IgG (P<0.05, P<0.01). The medium- and high-dose XEHT groups and the montelukast sodium showed down-regulated mRNA levels of IL-6, IL-1β, and TNF-α and the low-dose XEHT group showed down-regulated mRNA levels of IL-6 and TNF-α (P<0.05, P<0.01). Phospholipase D, mammalian target of rapamycin (mTOR), and epidermal growth factor receptor family of receptor tyrosine kinase (ErbB) signaling pathways were the common pathways predicted by both proteomics and network pharmacology. ConclusionProteomics combined with network pharmacology reveal that XEHT can ameliorate AC by regulating the phospholipase D, mTOR, and ErbB signaling pathways.
3.Clinical Characteristics and Influencing Factors of Rheumatoid Arthritis in Patients with Cold Dampness Obstruction Syndrome
Yanyu CHEN ; Yanqi LI ; Longxiao LIU ; Liubo ZHANG ; Tianyi LAN ; Nan ZHANG ; Cheng XIAO ; Yuan XU ; Qingwen TAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):140-146
ObjectiveTo study the clinical characteristics and influencing factors of rheumatoid arthritis (RA) in the patients with cold dampness obstruction syndrome. MethodsThe RA patients treated in the Department of Traditional Chinese Medicine and Rheumatology of the China-Japan Friendship Hospital from August 2022 to June 2024 were selected. The demographic information, clinical data, laboratory test results, and traditional Chinese medicine (TCM) symptom information were collected for syndrome differentiation, on the basis of which the characteristics and influencing factors of cold dampness obstruction syndrome were analyzed. ResultsA total of 258 RA patients were selected in this study, including 88 (34.1%) patients with cold dampness obstruction syndrome, 53 (20.5%) patients with dampness and heat obstruction syndrome, 31 (12.0%) patients with wind dampness obstruction syndrome, 29 (11.2%) patients with liver-kidney deficiency syndrome, 19 (7.4%) patients with Qi-blood deficiency syndrome, 14 (5.4%) patients with phlegm-stasis obstruction syndrome, 15 (5.8%) patients with stasis obstructing collateral syndrome and 9 (3.5%) patients with Qi-Yin deficiency syndrome. The patients were assigned into two groups of cold dampness obstruction syndrome and other syndromes. The group of cold dampness obstruction syndrome had lower joint fever, 28-tender joint count (TJC28), and 28-joint disease activity score (DAS28)-C-reactive protein (CRP) and higher central sensitization, cold feeling of joints, fear of wind and cold, cold limbs, and abdominal distention than the group of other syndromes (P<0.05). The binary logistic regression analysis showed that central sensitization (OR 5.749, 95%CI 2.116-15.616, P<0.001) and DAS28-CRP (OR 0.600, 95% CI 0.418-0.862, P=0.006) were the independent factors influencing cold dampness obstruction syndrome in RA. ConclusionCold dampness obstruction syndrome is a common syndrome in RA patients. It is associated with central sensitization, cold feeling of joints, abdominal distension and may be a clinical syndrome associated with central sensitization.
4.Differentiation and Treatment of Lipid Turbidity Disease Based on Theory of "Spleen Ascending and Stomach Descending"
Yun HUANG ; Wenyu ZHU ; Wei SONG ; Xiaobo ZHANG ; Xin ZHOU ; Lele YANG ; Tao SHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):244-252
Lipid turbidity disease is a metabolic disease featuring lipid metabolism disorders caused by many factors such as social environment, diet, and lifestyle, which is closely related to many diseases in modern medicine, such as hyperlipidemia, obesity, fatty liver, atherosclerosis, metabolic syndrome, and cardiovascular and cerebrovascular diseases, with a wide range of influence and far-reaching harm. According to the Huangdi Neijing, lipid turbidity disease reflects the pathological change of the body's physiologic grease. Grease is the thick part of body fluids, which has the function of nourishing, and it is the initial state and source of important substances in the human body such as brain, marrow, essence, and blood. Once the grease of the human body is abnormal, it can lead to lipid turbidity disease. The Huangdi Neijing also points out the physiological relationship between the transportation and transformation of body fluids and the rise and fall of the spleen and stomach, which can deduce the pathological relationship between the occurrence of lipid turbidity disease and the abnormal rise and fall of the spleen and stomach functions. Lipid turbidity disease is caused by overconsumption of fatty and sweet foods or insufficient spleen and stomach endowments, leading to disorders of the function of promoting clear and reducing turbidity in the spleen and stomach. This leads to the transformation of thick grease in body fluids into lipid turbidity, which accumulates in the body's meridians, blood vessels, skin pores, and organs, forming various forms of metabolic diseases. The research team believed that the pathological basis of lipid turbidity disease was the abnormal rise and fall of the spleen and stomach and the obstruction of the transfer of grease. According to the different locations where lipid turbidity stays, it was divided into four common pathogenesis types: ''inability to distinguish between the clear and turbid, turbid stagnation in the Ying blood'', ''spleen not rising clear, turbid accumulation in the vessels'', ''spleen dysfunction, lipid retention in the pores'', ''spleen failure to transportation and transformation, and grease accumulation in the liver''. According to the pathogenesis, it could be divided into four common syndromes, namely, turbid stagnation in the Ying blood, turbid accumulation in the vessels, lipid retention in the pores, and grease accumulation in the liver, and the corresponding prescriptions were given for syndrome differentiation and treatment, so as to guide clinical differentiation and treatment of the lipid turbidity disease.
5.Characteristics of public health emergencies in Jinhua City from 2014 to 2023
ZHANG Tao ; DU Zhiping ; WANG Zuoyi ; JIN Lü ; hua
Journal of Preventive Medicine 2025;37(1):69-72
Objective:
To investigate the characteristics of public health emergencies in Jinhua City, Zhejiang Province from 2014 to 2023, so as to provide the reference for prevention and control of public health emergencies.
Methods:
Data of public health emergencies and related information in Jinhua City from 2014 to 2023 were collected through Emergency Public Reporting System of Chinese Disease Prevention and Control Information System. Attack rates, and distribution of time, areas and places were descriptively analyzed.
Results:
A total of 276 public health emergencies were reported in Jinhua City from 2014 to 2023. There were 10 324 reported cases and 7 deaths, with an attack rate of 0.32%. There were 53 Ⅳ-level (19.20%) and 223 unclassified public health emergencies (80.80%). Infectious disease emergencies were predominant types, accounting for 97.83% (270 events). The three most common infectious disease emergencies were other infectious diarrhea (42.03%), influenza (21.01%) and COVID-19 (16.30%). The reported public health emergencies peaked in November and December, with 66 and 45 events reported, respectively. The three most counties (cities, districts) included Yiwu City, Wucheng District and Lanxi City, accounting for 24.28% (67 events), 18.48% (51 events) and 11.96% (33 events), respectively. School and preschool institutions were predominant places where public health emergencies occurred (198 events, 71.74%).
Conclusions
The public health emergencies in Jinhua City from 2014 to 2023 were Ⅳ-level and unclassified emergencies, and infectious disease emergencies were predominant. November and December were the peak reporting periods, and schools and preschool institutions were the main places where these events occurred.
6.Collection, storage and utilization of lung transplant tissue samples
Yixing LI ; Xue SHI ; Hongyi WANG ; Runyi TAO ; Ye SUN ; Ailing SU ; Liyan TONG ; Jinteng FENG ; Yanpeng ZHANG ; Shuo LI ; Yawen WANG ; Guangjian ZHANG
Organ Transplantation 2025;16(1):147-155
After continuous development and improvement, lung transplantation has become the preferred means to treat a variety of benign end-stage lung diseases. However, the field of lung transplantation still faces many challenges, including shortage of donor resources, preservation and maintenance of donor lungs, and postoperative complications. Lung tissue samples removed after lung transplantation are excellent clinical resources for the study of benign end-stage lung disease and perioperative complications of lung transplantation. However, at present, the collection, storage and utilization of tissue samples after lung transplantation are limited to a single study, and unified technical specifications have not been formed. Based on the construction plan of the biobank for lung transplantation in the First Affiliated Hospital of Xi'an Jiaotong University, this study reviewed the practical experience in the collection, storage and utilization of lung transplant tissue samples in the aspects of ethical review, staffing, collection process, storage method, quality control and efficient utilization, in order to provide references for lung transplant related research.
7.Effect and Mechanism of Wulingsan Decoction in Protecting Blood Brain Barrier and Ameliorating Cerebral Edema after Intracerebral Hemorrhage in Mice
Damei TAO ; Huihong LI ; Xiaoqing ZHENG ; Yunfei DENG ; Wei WEI ; Xiehua XUE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):1-9
ObjectiveTo study the effect of Wulingsan on cerebral edema after intracerebral hemorrhage (ICH) in mice and explore the treatment mechanism. MethodsThe mouse model of ICH was established by injection of collagenase into the caudate nucleus. Mice were randomly assigned into the following groups: sham, ICH, intervention before modeling with low-dose and high-dose (3.69, 11.07 g·kg-1, respectively) Wulingsan, and intervention after modeling with high-dose Wulingsan. The modified neurological severity score (mNSS) was recorded, and the small animal MRI T2 sequential scanning was performed to measure the volume of cerebral hemorrhage after the modeling of ICH in each group. The Y-maze test, open field test, and Morris water maze test were conducted to evaluate the neurological behaviors of mice in each group. Hematoxylin-eosin staining was employed to observe the pathological changes in the brain tissue. Immunohistochemistry was employed to observe the expression of aquaporin 4 (AQP4), neuronal nuclei (NeuN), and glial fibrillary acidic protein (GFAP) in the brain tissue. Western blot was employed to determine the protein levels of AQP4, Claudin-5, and zonula occludens-1 (ZO-1) in the hematoma area. ResultsCompared with the sham group, the ICH group showed increases in the mNSS, the cerebral hemorrhage volume, and the escape latency in the Morris water maze test (P<0.01), decreases in the times of touching the platform and times of entering the quadrant where the platform was located in the Morris water maze test, and reductions in the spontaneous alternation rate in the Y-maze test and the ratio of distance of center travel to total travel distance in the open field test (P<0.01). Moreover, pathological changes such as cell disarrangement, cell space enlargement, and cell swelling were observed in the ICH group. Immunohistochemistry results showed that the ICH group had higher proportions of AQP4- and GFAP-positive cells and lower proportion of NeuN-positive cells than the sham group (P<0.01). Compared with the sham group, the ICH group showed an up-regulated protein level of AQP4 and down-regulated protein levels of Claudin-5 and ZO-1 (P<0.01). Compared with the ICH group, the intervention with Wulingsan decreased the mNSS, the volume of cerebral hemorrhage, and the escape latency in the Morris water maze test (P<0.05, P<0.01), while increasing the times of touching the platform and times of entering the quadrant where the platform was located in the Morris water maze test, the spontaneous alternation rate in the Y-maze test, and the ratio of distance of center travel to total travel distance in the open field test (P<0.05, P<0.01). Furthermore, the intervention with Wulingsan alleviated the pathological changes in the brain tissue after ICH, decreased the proportion of AQP4- and GFAP-positive cells (P<0.01), increased the proportion of NeuN-positive cells (P<0.01), down-regulated the protein level of AQP4 (P<0.01), and up-regulated the protein levels of Claudin-5 and ZO-1 (P<0.01). ConclusionThe intervention with Wulingsan could reduce the neural function score and the cerebral hemorrhage volume, up-regulate the expression of Claudin-5 and ZO-1, and down-regulate the expression of AQP4 to ameliorate the neurological function defect and cerebral edema after ICH, thereby protecting the brain.
8.Application of Zebrafish in Quality Evaluation of Traditional Chinese Medicine
Jiaqi LI ; Xiaolu CHEN ; Jiarui WU ; Xiaoyu TAO ; Qiqi FAN ; Shengyun DAI ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):58-70
As a new type of model organism, zebrafish is gradually gaining prominence in the field of scientific research. The unique biological characteristics and advantages of zebrafish make them play an increasingly important role in the quality evaluation of traditional Chinese medicine. Compared with other common experimental animals, zebrafish have a fast reproductive and growth speed and high embryo transparency, making them an ideal model for evaluating the quality of traditional Chinese medicine. This provides a new perspective and method for research on traditional Chinese medicine. With the growing global interest in traditional Chinese medicine, it has become crucial to find scientific and accurate methods to evaluate the quality and effectiveness of traditional Chinese medicine. The introduction of the zebrafish model has brought new breakthroughs in the quality evaluation of traditional Chinese medicine. To further promote the application of zebrafish in evaluating the quality of traditional Chinese medicine, this article systematically searched and sorted out the previous studies related to the application of zebrafish for this purpose since 2023. The commonly used disease models and indicators of zebrafish in evaluating the effectiveness of traditional Chinese medicine, as well as the mechanism of zebrafish in exploring the active ingredients of traditional Chinese medicine, were primarily reviewed. The application of zebrafish in evaluating the safety of traditional Chinese medicine and the typical examples in ensuring the quality of traditional Chinese medicine were demonstrated. The limitations encountered by zebrafish models in evaluating the quality of traditional Chinese medicine were highlighted. The resolution of these problems will help further improve the accuracy and reliability of zebrafish in evaluating the quality of traditional Chinese medicine. The article discussed the evaluation of effectiveness, safety, and quality control of zebrafish applied in traditional Chinese medicine, so as to provide a reference for establishing standards for traditional Chinese medicine and promoting its modernization in the future.
9.Hepatotoxicity and Anti-osteoporotic Mechanism of Evodiamine
Jiaqi LI ; Xiaolu CHEN ; Xiaoyu TAO ; Shan LU ; Qiqi FAN ; Jiarui WU ; Chongjun ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):79-86
ObjectiveBased on the zebrafish model, the hepatotoxicity and anti-osteoporotic activity of evodiamine (EVO) were studied. The mechanism of EVO in treating osteoporosis was explored by using network pharmacology and real-time polymerase chain reaction(Real-time PCR). MethodsThree days after fertilization (3 dpf), zebrafish were randomly selected and exposed to different concentrations of EVO solution for 96 hours. The mortality rate of zebrafish at different concentrations was calculated at the exposure endpoint, and a "dose-toxicity" curve was drawn. The 10% lethal concentration (LC10) was calculated. Liver phenotype, acridine orange staining, and pathological tissue sections of liver-transgenic zebrafish [CZ16 (gz15Tg.Tg (fabp 10a: ds Red; ela31: EGFP))] were used to confirm hepatotoxicity of EVO. On this basis, prednisolone was used to create a model of osteoporosis in zebrafish. The skull development, area of the skull stained by alizarin red, and cumulative optical density were used as indicators to evaluate the anti-osteoporotic activity of EVO in a safe dose. Based on network pharmacology, the mechanism of action of EVO in the treatment of osteoporosis was predicted and verified through Real-time PCR. ResultsThe LC10 of EVO on zebrafish (7 dpf) was determined to be 0.4 mg·L-1. Compared with the control group, sublethal concentrations (
10.Weichang'an Prescription-containing Serum Induces Ferroptosis of Gastric Cancer MKN-45 Cells
Xin LI ; Jinzu YANG ; Jianxin QIAN ; Li TAO ; Ling CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):132-139
ObjectiveTo observe the effect of Weichang'an prescription-containing serum on ferroptosis of human gastric cancer cells and explore the possible mechanism. MethodsSD rats were administrated with 18, 36, 72 g·kg-1·d-1 Weichang'an prescription by gavage for preparation of serum samples containing different doses of Weichang'an prescription, which were then used to treat MKN-45 cells. The cell proliferation was examined by the cell counting kit-8 (CCK-8). In addition, inhibitors of apoptosis, necroptosis, and ferroptosis were added, and the survival of the cells treated with the serum samples was observed. The fluorescent probe dichlorodihydrofluorescein diacetate (DCF-DA) and the lipid peroxidation sensor C11-BODIPY were employed to detect the intracellular levels of reactive oxygen species (ROS) and lipid peroxidation, respectively. The levels of ferrous ion (Fe2+), glutathione (GSH), and malondialdehyde (MDA) were detected by enzyme-linked immunosorbent assay (ELISA). Real-time PCR and Western blotting were employed to determine the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), aldo-keto reductase family 1 member B1 (AKR1B1), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4), signal transducer and activator of transcription 3 (STAT3), and mitogen-activated protein kinase (MAPK). ResultsCompared with the blank group, Weichang'an prescription-containing serum decreased the viability of MKN-45 cells (P<0.05, P<0.01) in a time- and dose-dependent manner. Compared with the Weichang'an prescription group, the apoptosis inhibitor+Weichang'an prescription group and the ferroptosis inhibitor+Weichang'an prescription group showed increased cell viability (P<0.05, P<0.01). Compared with the blank group, Weichang'an prescription elevated the levels of ROS, lipid peroxidation, and intracellular Fe2+ and MDA (P<0.05, P<0.01) and lowered the level of GSH (P<0.05, P<0.01) in a dose-dependent manner. Compared with the blank group, Weichang'an prescription down-regulated the mRNA and protein levels of Nrf2, AKR1B1, and GPX4 (P<0.05, P<0.01) and up-regulated the mRNA and protein levels of ACSL4 (P<0.05, P<0.01) in a dose-dependent manner. Compared with the blank group, Weichang'an prescription down-regulated the protein levels of p-STAT3 and p-ERK (P<0.05, P<0.01) in a dose-dependent manner. ConclusionThe Weichang'an prescription-containing serum can promote the ferroptosis and inhibit the proliferation of MKN-45 cells by regulating the STAT3 and MAPK pathways.


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