1.Chufeng Yisuntang Ameliorates PM2.5-induced Dry Eye via ROS/p38 MAPK Signaling Pathway
Yuan ZHONG ; Pan ZHAO ; Shi TAN ; Yu TANG ; Dongdong LI ; Lihao CHEN ; Jun PENG ; Qinghua PENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):191-200
ObjectiveTo establish a mouse model of particulate matter 2.5 (PM2.5)-induced dry eye and investigate whether Chufeng Yisuntang can ameliorate the PM2.5-induced ocular surface damage by regulating the reactive oxygen species (ROS)/p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. MethodsSixty 8-week-old male C57BL/6J mice were used. Ten were randomly selected as the control group. The remaining 50 mice received topical instillation of 1 drop (0.1 mL) of 5 g·L-1 PM2.5 suspension in both eyes, four times daily. Successfully modeled mice were randomized into four groups (n=10): Model, p38 MAPK inhibitor, Chufeng Yisuntang, and combination (Chufeng Yisuntang at 7.3 g·kg-1 + p38 MAPK inhibitor SB203580 at 5 mg·kg-1). Chufeng Yisuntang was administered via gavage, and the inhibitor group via intraperitoneal injection. The control and model groups received equal volumes of distilled water by gavage. All treatments lasted for 4 weeks. General conditions were dynamically observed. Tear secretion, tear film break-up time, and corneal fluorescein staining were assessed. After intervention for 4 weeks, hematoxylin and eosin (HE) staining was used to examine the histopathological changes. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure serum levels of ROS, malondialdehyde (MDA), superoxide dismutase (SOD) 1, and SOD2. Western blot and Real-time PCR were employed to determine the protein and gene levels, respectively, of p38 MAPK, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteinyl aspartate-specific proteinase-3 (Caspase-3) in the corneal tissue. ResultsCompared with the control group, the model group exhibited reduced tear secretion volume and tear film breakup time, along with increased corneal fluorescein staining scores (P<0.01). Compared with the model group, the Chufeng Yisuntang group, p38 MAPK inhibitor group, and combination group demonstrated increased tear secretion volume and tear film breakup time, along with decreased corneal fluorescein staining scores (P<0.01). HE staining revealed that compared with the control group, the model group exhibited marked increases in corneal epithelial cell layers and epithelial thickness, along with reduced meibomian gland acini and intensely stained, densely packed nuclei around the acini. Compared with the model group, the Chufeng Yisuntang group, p38 MAPK inhibitor group, and combination group showed intact corneal structure, improved cell morphology, and reduced damage severity. ELISA revealed elevated ROS and MDA levels (P<0.01) and decreased SOD1 and SOD2 levels (P<0.01) in the model group compared with the control group. Compared with the model group, Chufeng Yisuntang, p38 MAPK inhibitor, and the combination lowered ROS and MDA levels (P<0.01), while raising SOD1 and SOD2 levels (P<0.05, P<0.01). Western blot revealed that compared with the control group, the model group exhibited increased protein levels of p38 MAPK, Bax, and Caspase-3 (P<0.01) and reduced protein level of Bcl-2 (P<0.01). Compared with the model group, Chufeng Yisuntang, p38 MAPK inhibitor, and the combination down-regulated the protein levels of p38 MAPK, Bax, and Caspase-3 (P<0.01), while up-regulating the protein level of Bcl-2 (P<0.01). Compared with the Chufeng Yisuntang group, the combination group exhibited decreased protein levels of p38 MAPK, Bax, and Caspase-3 (P<0.01) and increased protein level of Bcl-2 (P<0.01). Real-time PCR revealed that compared with the control group, the model group exhibited upregulated mRNA levels of p38 MAPK, Bax, and Caspase-3 (P<0.01), and downregulated mRNA level of Bcl-2 (P<0.01). Compared with the model group, Chufeng Yisuntang, p38 MAPK inhibitor, and the combination down-regulated the mRNA levels of p38 MAPK, Bax, and Caspase-3 (P<0.01), while up-regulating the mRNA level of Bcl-2 (P<0.05, P<0.01). Compared with the Chufeng Yisuntang group, the combination group exhibited decreased mRNA levels of p38 MAPK, Bax, and Caspase-3 expression (P<0.05, P<0.01) and increased mRNA level of Bcl-2 (P<0.01). ConclusionChufeng Yisuntang may partially protect against PM2.5-induced corneal injury by inhibiting the ROS/p38 MAPK pathway, enhancing antioxidant defense, and reducing epithelial apoptosis.
2.Proanthocyanidins activate Nrf2/GPX4 pathway to inhibit ferroptosis and exert protective effects on dry ARMD
International Eye Science 2026;26(7):1116-1126
AIM: To investigate whether proanthocyanidins(PAC)can ameliorate ferroptosis in retinal pigment epithelium(RPE)cells via the Nrf2/GPX4 pathway, and to evaluate its protective effect in dry age-related macular degeneration(ARMD). METHODS: The protective effects of PAC against ferroptosis were evaluated using an in vitro model of H2O2-stimulated human RPE cells(ARPE-19)and an in vivo dry ARMD animal model induced by sodium iodate(NaIO3). CCK-8 assay was applied to assess cell viability. Calcein-AM/PI staining was applied to determine the level of cell death. Lipid peroxidation(LPO)levels were measured using a lipid peroxidation assay kit. CM-H2DCFDA was used to detect the level of reactive oxygen species(ROS)in cells. MitoSOX probe was employed to measure the mitochondrial ROS level. JC-1 staining was used to evaluate the changes in mitochondrial membrane potential. Western Blot was performed to detect the expression levels of proteins(Nrf2, GPX4, and HO-1)related to the Nrf2 pathway. Retinal flat mounts were used to evaluate the structure and function of RPE. Hematoxylin and eosin(H & E)staining was applied to assess the morphological changes in rat retinas. Rhodopsin and opsin staining was used to evaluate visual function in the retina. TUNEL staining was employed to detect apoptosis in retinal cells. RESULTS: H2O2 exacerbates ferroptosis in ARPE-19 cells, as evidenced by elevated levels of Fe2+, ROS, and LPO. PAC preconditioning ameliorates mitochondrial function, reduces intracellular Fe2+, ROS, and LPO levels, and suppresses ferroptosis in RPE cells via activation of the Nrf2/GPX4 signaling pathway. In a NaIO3-induced dry ARMD model, both PAC and the ferroptosis inhibitor Fer-1 reverse the downregulation of retinal Nrf2/GPX4 expression and attenuate retinal cell death.CONCLUSION:PAC inhibits ferroptosis via the Nrf2/GPX4 pathway, offering a novel potential therapeutic strategy for retinal degeneration.
3.Horticultural Therapy Combined with Intradermal Needling for Patients with Generalized Anxiety Disorder of Liver Depression Transforming into Fire Syndrome Under Transcranial Magnetic Stimulation and Psychological Therapy:Clinical Observation of 60 Cases
Wanyun ZHANG ; Jiayi YAN ; Qingyi QIU ; Yumei PENG ; Xiaoling ZHONG ; Jinwen ZHANG ; Rundong TANG ; Miao WU ; Dan HU ; Guang SU
Journal of Traditional Chinese Medicine 2025;66(1):50-58
ObjectiveTo observe the clinical effectiveness of horticultural therapy involving the planting of Chinese medicinal herbs (mint and lily potted plants) combined with intradermal needling therapy for generalized anxiety disorder (GAD) of liver depression transforming into fire syndrome under transcranial magnetic stimulation and basic psychological therapy, and to explore the possible mechanisms of action. MethodsA total of 180 patients with GAD of liver depression transforming into fire syndrome were randomly divided into three groups, horticultural therapy group, intradermal needling group, and horticultural therapy+intradermal needling group, with 60 patients in each. All groups received basic treatment including basic psychological therapy and transcranial magnetic stimulation. The horticultural therapy group received horticultural therapy in addition to the basic treatment; the intradermal needling group received intradermal needling therapy once a week for 8 weeks in addition to the basic treatment; the horticultural therapy+intradermal needling group received both horticultural therapy and intradermal needling therapy, following the same procedures and duration. Hamilton Anxiety Rating Scale (HAMA), Self-Rating Anxiety Scale (SAS), and Pittsburgh Sleep Quality Index (PSQI) scores were assessed at baseline and after 2, 4, 6, and 8 weeks of treatment. Serum levels of adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were measured before treatment and after 8 weeks of treatment. Motor-evoked potential (MEP) baseline levels were recorded before treatment, and MEP amplitude ratios were compared after 1 week and 8 weeks of treatment. Clinical effectiveness and safety were evaluated after 8 weeks of treatment. Pearson correlation analysis was used to examine the relationships between serum ACTH and CORT levels, MEP amplitude, and anxiety. ResultsIn the horticultural therapy group and intradermal needling group, HAMA, SAS and PSQI scores after 4, 6, and 8 weeks treatment were lower than baseline scores (P<0.05). In the horticultural therapy+intradermal needling group, these scores showed a significant decline starting after 2 weeks treatment and continuing through 8 weeks after treatment (P<0.05). The HAMA, SAS, and PSQI scores in the horticultural therapy+intradermal needling group were significantly lower than those in the other two groups after 2, 4, 6, and 8 weeks treatment (P<0.05). After 8 weeks of treatment, serum CORT and ACTH levels in the horticultural therapy+intradermal needling group were significantly lower than baseline levels (P<0.05) and were also lower than those in the horticultural therapy group and intradermal needling group at the same time point (P<0.01). When comparing the level after 8 weeks treatment to that after 1 week treatment, under PAS10 stimulation, the MEP amplitude ratio in the intradermal needling group decreased at 30 minutes, while in the horticultural therapy+intradermal needling group, the MEP amplitude ratio decreased at all time points (P<0.05 or P<0.001); under PAS25 stimulation, the MEP amplitude ratio in the horticultural therapy group increased at 20 minutes, and in the intradermal needle group at 10 minutes (P<0.05). In the horticultural therapy+intradermal needling group, the MEP amplitude ratio increased significantly at all time points after treatment (P<0.001). The cure rate in the horticultural therapy+intradermal needling group (74.14%, 43/58) was significantly higher than that in the horticultural therapy group (30.00%, 18/60) and the intradermal needling group (48.28%, 28/58, P<0.05). Correlation analysis revealed that serum ACTH and CORT levels were positively correlated with HAMA scores (r = 0.488, P<0.01; r = 0.428, P<0.01). Following PAS10 intervention, the MEP amplitude ratio was positively correlated with HAMA scores (r = 0.458, P<0.01), whereas after PAS25 intervention, the MEP amplitude ratio was negatively correlated with HAMA scores (r = -0.562, P<0.01). ConclusionHorticultural therapy combined with intradermal needling treatment, under transcranial magnetic stimulation and basic psychological therapy, demonstrates significant clinical effectiveness in patients with GAD of liver depression transforming into fire syndrome. Its mechanism of action may be related to the regulation of hyperactivation of the hypothalamic-pituitary-adrenal (HPA) axis and the reduction of cortical excitability.
4.Research progress of meibomian gland dysfunction-related dry eye
Jianbo ZHONG ; Guoqiang ZENG ; Yi ZHANG ; Xiaoyan DOU ; Wanmei TANG ; Kunling CHEN ; Li CAI
International Eye Science 2025;25(2):259-263
In recent years, with the endless emergence of meibomian gland dysfunction(MGD)diagnostic equipment, rich treatment methods, and in-depth clinical and basic research on MGD at home and abroad, the understanding of MGD has entered a new stage. MGD-related dry eye is considered to be the main cause of lipid abnormal dry eye, and its occurrence and development is a chronic and multi-factorial pathological process. This article reviews the pathogenesis, imaging analysis and clinical treatment progress of MGD-related dry eye, in order to provide scientific evidence and ideas for clinical diagnosis and therapy of MGD-related dry eye.
5.Polymorphism and Tissue Expression Analysis of TYR and MC1R Genes in Guinea Pigs with Different Coat-Color Phenotypes
Yingen TANG ; Yaxian FENG ; Min ZHONG ; Zhen WEI ; Lie WANG ; Diwen LIU
Laboratory Animal and Comparative Medicine 2025;45(1):21-29
Objective To explore the polymorphism of tyrosinase (TYR) and melanocortin 1 receptor (MC1R) genes and their mRNA expression levels in relation to coat-color phenotypes in guinea pigs, providing genetic markers for locating dominant traits in guinea pigs. Methods A total of 57 self-bred ordinary-level guinea pigs were selected and divided into three groups based on coat color: white (n=22), variegated (n=22) and black (n=13). The guinea pigs were euthanized with an overdose of pentobarbital sodium via intraperitoneal injection. DNA was then extracted from the dorsal skin tissue. Polymorphism in the coding sequence (CDS) of the exons of the TYR and MC1R genes in each group was detected by cloning and sequencing. The mRNA expression of the two genes in skin tissues was detected by real-time fluorescent quantitative PCR to investigate the relationship between these genes and guinea pig coat color. Results A single nucleotide polymorphism (SNP) site was found in the CDS region of TYR exon Ⅰ, where the base A was replaced by G. All white guinea pigs had the G/G genotype for TYR, while no deep-colored (variegated and black) guinea pigs exhibited the G/G genotype for TYR. Most deep-colored guinea pigs had the A/A genotype, and a few had A/G genotype. The A/A genotype frequency in black guinea pigs was higher than in variegated guinea pigs. A 2 760 bp sequence deletion was identified in the exon of the MC1R gene, marked as the - gene, with non-deleted samples marked as N gene. Most white guinea pigs had the -/- genotype for MC1R, variegated guinea pigs mainly had the -/N genotype, and black guinea pigs mainly had the N/N genotype, with a few showing the -/N. The TYR gene expression level was higher in white guinea pigs, lower in variegated guinea pigs, and intermediate in black guinea pigs, but there was no significant difference among the three groups (P>0.05). The MC1R gene expression level in white guinea pigs was extremely low, while both variegated and black guinea pigs showed significantly higher levels than white guinea pigs (P<0.01). Black guinea pigs showed significantly higher levels than variegated guinea pigs (P<0.05). ConclusionThe TYR and MC1R genes synergistically regulate coat color of guinea pigs. The G-site mutation in the TYR gene may lead to albinism, and the change of N-site in the MC1R gene affects the depth of the coat color.
6.Assessing distribution characteristics and clinical significance of vertebral fractures in patients with osteoporosis based on whole spine MRI
Jiajun ZHOU ; Fei MA ; Yebo LENG ; Shicai XU ; Baoqiang HE ; Yang LI ; Yehui LIAO ; Qiang TANG ; Chao TANG ; Qing WANG ; Dejun ZHONG
Chinese Journal of Tissue Engineering Research 2025;29(9):1883-1889
BACKGROUND:Osteoporotic vertebral fractures are the most common complication in patients with osteoporosis.As a new imaging technique,spine magnetic resonance imaging(MRI)is much more sensitive than X-ray film in the diagnosis of osteoporotic vertebral fractures.However,total spine MRI is costly and takes a long time to scan.Therefore,there is no consensus on whether all patients with osteoporotic vertebral fractures need to undergo total spine MRI scan and which patients need to undergo total spine MRI. OBJECTIVE:To analyze the distribution characteristics of vertebral fractures and explore their clinical significance by observing the whole spine MRI data of osteoporotic vertebral fractures patients. METHODS:Data of cases and MRI images of all patients diagnosed with fresh osteoporotic vertebral fractures who visited the Department of Orthopedics,Affiliated Hospital of Southwest Medical University from August 2018 to September 2022 were retrospectively analyzed.903 patients were included in the study based on inclusion and exclusion criteria.General information(age,gender,and body mass index),medical history characteristics(duration of illness,history of trauma surgery,percussion pain area,and pain score)were collected.The characteristics of vertebral fractures were analyzed through whole spine magnetic resonance imaging.Firstly,based on the number of vertebral fractures in patients,they were divided into the single vertebral fracture group(484 cases)and the multi-vertebral fracture group(419 cases),and the differences were analyzed between the two groups.Then,based on whether the farthest interval between the fractured vertebrae was greater than or equal to 5,the multi vertebral fracture group was further divided into two subgroups.Among them,Group A(the farthest interval between the fractured vertebrae was less than 5)contained 306 cases;Group B(with the farthest interval between fractured vertebral bodies greater than 5)included 113 cases.The differences were analyzed between two subgroups. RESULTS AND CONCLUSION:(1)Among 903 patients,419 patients(46.4%)had more than two fractured vertebrae.There were 654 patients(72.4%)with thoracolumbar fractures,and 54 patients(6%)with fractures in the thoracic plus lumbar region and the entire thoracic to lumbar region.In group B,96.5%of patients had multiregional percussion pain.(2)Compared with the patients in the single vertebral fracture group and the multi-vertebral fracture group,there were significant differences in bone mineral density,whether the medical history was greater than or equal to 1 month,the history of low energy injury,and the distribution and number of axial percussion pain areas in the spine during physical examination between the two groups(P<0.05).Age,gender,body mass index,whether there was underlying disease,pain visual analog scale score,whether there was a history of elderly thoracolumbar fracture,and whether there was a history of thoracolumbar surgery,and the number of fractured vertebrae had no statistical significance(P>0.05).(3)There were statistically significant differences between the Groups A and B in bone mineral density,the distribution and quantity of percussion pain area,and the history of low energy injury(P<0.05).There were no significant differences in age,gender,history of old fractures,visual analog scale score,body mass index,whether the medical history was longer than or equal to 1 month,history of underlying diseases,and history of thoracolumbar surgery between the two groups(P>0.05).(4)Patients with multiple low-energy trauma history,history of more than 1 month,multiple percussion pain,and the lower bone mineral density should be alert to the occurrence of multiple vertebral fracture and jump fracture.We recommend the whole spinal MRI for these patients.
7.Visualization analysis of knowledge map and trends in glymphatic system research
Xingyi MA ; Huijing LI ; Juan LI ; Dongling ZHONG ; Yuan ZHANG ; Hong TANG ; Yuxi LI ; Rongjiang JIN
Chinese Journal of Tissue Engineering Research 2025;29(23):5051-5060
BACKGROUND:The clearance pathway of metabolic waste in the brain is crucial for maintaining neural homeostasis.The accumulation of metabolic waste disrupts this equilibrium,representing a common pathological feature of many central nervous system diseases.In recent years,research focusing on the glymphatic system has emerged as a hotspot in the nervous system field.OBJECTIVE:To construct a knowledge map of glymphatic system research,visually analyze the current state of research,main hotspots,and future development trends within this area.METHODS:Utilizing CiteSpace,VOSviewer,and the Bibliometrix package in the R language environment,this study conducted an in-depth visual analysis of glymphatic system-related literature from the Web of Science Core Collection database,spanning from January 2012 to March 2024.This analysis included authors,institutions,countries,journals,keywords,and co-citation frequencies.RESULTS AND CONCLUSION:A total of 687 related articles were included in this study.The number of publications in this field increased year by year,showing an explosive growth trend in the past three years.The countries,institutions,and authors with the largest number of publications in this research field were the United States,University of Rochester,and Professor Maiken Nedergaard.The journal with the highest number of publications was JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM.The high-frequent and high-central keywords mainly focused on the mechanism such as cerebrospinal fluid hydrodynamics,neurological diseases such as Alzheimer's disease,and imaging techniques such as diffusion tensor imaging.The literature with the highest co-citation frequency was a classic review of the glymphatic system.The above results show that the research of glymphatic system is an emerging and active field,which has attracted wide attention at home and abroad and gradually expand from theoretical research to clinical practice.
8.Analysis of the subjective and objective effects of family care level and disease acceptance among maintenance hemodialysis patients and their caregivers
Yahui ZHANG ; Li ZHANG ; Yuxin ZHONG ; Kaihui DENG ; Le TANG ; Yidan FENG
Chinese Journal of Practical Nursing 2025;41(14):1056-1063
Objective:The actor-partner interdependence model was used to explore the subject-object effect of family care on disease acceptance in maintenance hemodialysis (MHD) patients and their primary caregivers, and to provide empirical research support for formulating intervention strategies to improve the quality of life of MHD patients and their primary caregivers at the dyadic coping perspective.Methods:This study was a cross-sectional study. A convenience sampling method was used to select 223 pairs of MHD patients and their primary caregivers who received treatment at three hospitals from December 2023 to May 2024, namely the Second Affiliated Hospital of Guizhou Medical University, Qiandongnan People′s Hospital and Qiandongnan Traditional Chinese Medicine Hospital. The General Information Questionnaire, Family APGAR Index (APGAR) and the Acceptance of Illness Scale (AIS) were used to conduct the survey. An actor-partner interdependence model of the impact of family care levels on disease acceptance was constructed.Results:There were 223 patients with MHD, 134 males and 89 females, aged (48.41 ± 14.41) years. Among the 223 primary caregivers of MHD patients, 83 were males and 140 were females, aged (49.44 ± 12.40) years. The scores of APGAR and AIS for MHD patients were (7.92 ± 1.94), (20.45 ± 4.66) points, and (8.16 ± 1.67), (21.86 ± 3.54) points for their primary caregivers. There were statistically significant differences in scores ( t = - 2.28, - 7.69, both P<0.05). The results of correlation analysis showed that there was a significant positive correlation between the degree of family care of MHD patients and the disease acceptance of MHD patients, the family care of primary caregivers, and the disease acceptance of primary caregivers ( r = 0.454, 0.625, 0.515, all P<0.05). There was a significant positive correlation between the disease acceptance of MHD patients and the family care of their primary caregivers, the disease acceptance of primary caregivers, and the family care of primary caregivers and the disease acceptance of primary caregivers ( r = 0.442, 0.813, 0.495, all P<0.05). In terms of the actor effect, the level of family care positively influenced the disease acceptance for both MHD patients and their primary caregivers ( β = 0.715, 0.603, both P<0.001), the actor effect was significant. In terms of partner effect, there was a positive correlation between the family care levels of MHD patients and their primary caregivers and the disease acceptance of the other party ( β = 0.628, 0.725, both P<0.001), the partner effect was significant. Conclusions:There is an interactive effect between the disease acceptance of maintenance hemodialysis patients and their primary caregivers and their levels of family care.
9.Application of combined stent and catheter thrombectomy technique in severe cerebral venous sinus thrombosis
Lin XIANG ; Genlong ZHONG ; Yanan TANG ; Weikang CHEN
Chinese Journal of Primary Medicine and Pharmacy 2025;32(8):1132-1136
Objective:To investigate the effects of combined stent and catheter thrombectomy technique on severe cerebral venous sinus thrombosis.Methods:A single-center, prospective, randomized controlled study was conducted involving 78 patients with severe cerebral venous sinus thrombosis who were treated at Lishui People's Hospital from January 2019 to December 2023. Using the random number table method, these patients were divided into two groups: an experimental group ( n = 39) and a control group ( n = 39). The control group received heparin anticoagulation treatment, while the experimental group underwent stent placement combined with catheter thrombectomy. The patency rate of occluded venous sinuses, Glasgow Coma Scale scores, Glasgow Outcome Scale scores, overall prognosis, and the incidence of adverse events were compared between the two groups. Results:After treatment and at 90 days post-treatment, the patency rate of occluded venous sinuses in the experimental group was 89.7% (35/39) and 79.5% (31/39), respectively. These were significantly higher than those in the control group [71.8% (28/39), 56.4% (22/39), χ2 = 4.04, 4.77, both P < 0.05]. Both the Glasgow Coma Scale scores and Glasgow Outcome Scale scores significantly increased in both groups after treatment, with the experimental group showing higher scores than the control group (all P < 0.05). At 30 and 90 days after treatment, the proportion of patients with a modified Rankin Scale score of ≤ 2 in the experimental group was 87.2% (34/39) and 79.5% (31/39), respectively. These were significantly higher than those in the control group [66.7% (26/39), 53.8% (21/39), χ2 = 4.62, 5.77, both P < 0.05]. There was no significant difference in the total incidence of adverse events between the two groups ( P > 0.05). Conclusions:The stent combined with catheter thrombectomy technique exhibits a considerable application value for patients with severe cerebral venous sinus thrombosis. This approach can enhance the vascular recanalization rate, promote disease recovery, improve patient prognosis, and is considered safe.
10.Study on synergistic promotion of ferroptosis in human hypertrophic scar fibroblasts by erastin combined with shikonin
Jian-jun WANG ; Yan-hua WANG ; Yu-ting TANG ; Jing-yi ZHANG ; Fang MA ; Xi HE ; Hui-xia YANG ; Qi-peng ZHAO ; Zhi-gang BAI ; Yin-ju HAO ; Gui-zhong LI ; Yi-deng JIANG ; Jiang-yong SHEN
Chinese Pharmacological Bulletin 2025;41(2):268-276
Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50%).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P<0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P<0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P<0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P<0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

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