1.Modified Xiaoyaosan Alleviates Neuronal Dysfunction in Rat Model of Post-myocardial Infarction Depression by Regulating Mitochondrial Quality Control Through Drp1/PINK1/Parkin Signaling Pathway
Zhen ZHONG ; Dongsheng WEI ; Xinyue XIONG ; Lin LI ; Mingli YAO ; Xinnuan SHI ; Youming JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):20-31
ObjectiveTo investigate the effects of modified Xiaoyaosan (JJXYS) on behavioral abnormalities and hippocampal mitochondrial quality control (MQC) in the rat model of post-myocardial infarction depression (PMD) and preliminarily explore its potential mechanism. MethodsA rat model of PMD was established by left anterior descending coronary artery ligation combined with chronic unpredictable mild stress (CUMS). Rats were randomized into a control group, a model group, a fluoxetine (FLX, 10 mg·kg-1) group, and low-, medium-, and high-dose JJXYS (JJXYS-L/M/H, 1.12, 2.24, 4.48 g·kg-1, respectively) groups. Depressive-like behaviors were evaluated by body weight monitoring, sucrose preference test, open field test, and forced swimming test. Hematoxylin-eosin staining and Nissl staining were used to observe hippocampal histomorphology and neuronal changes. Enzyme-linked immunosorbent assay was conducted to determine the serum levels of 5-hydroxytryptamine (5-HT), dopamine (DA), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). The mRNA levels of MQC-related genes including peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1α), nuclear respiratory factor 1 (Nrf1), and transcription factor A, mitochondrial (TFAM) in the hippocampal tissue were measured by real-time PCR. The expression of proteins related to the dynamin-related protein 1 (Drp1)/PTEN-induced putative kinase 1 (PINK1)/Parkin signaling pathway was determined by Western blot. ResultsCompared with the control group, the model group showed restricted body weight gain, aggravated depressive-like behaviors, declined serum 5-HT and DA levels, evident hippocampal neuronal damage and reduced Nissl bodies, as well as downregulated expression of MQC-related genes and proteins (P<0.05). Compared with the model group, both FLX and JJXYS alleviated the above changes to varying degrees. Moreover, the JJXYS-M and JJXYS-H groups showed more pronounced effects, improving behavioral performance, restoring 5-HT and DA levels, alleviating hippocampal pathological injury, and upregulating the expression of PGC-1α/Nrf1/TFAM mRNA and Drp1/PINK1/Parkin signaling pathway-related proteins (P<0.05). ConclusionJJXYS can significantly alleviate depressive-like behaviors and neurotransmitter imbalance in the rat model of PMD by regulating hippocampal MQC and upregulating the Drp1/PINK1/Parkin-related pathway. This study provides experimental evidence for the intervention of PMD with JJXYS.
2.Analysis of Blood-absorbed Components and Their Metabolic Differences of Xiebaisan in Normal and Chronic Bronchitis Mice Based on UPLC-Q-Exactive Orbitrap MS
Peng PENG ; Jiaxin LI ; Xinyue YANG ; Fangle LIU ; Chenchen ZHU ; Chaozhan LIN ; Yufeng YAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):219-227
ObjectiveThis study aims to systematically analyze the blood-absorbed components and metabolic profiles of Xiebaisan(XBS) in normal and chronic bronchitis (CB) mice using ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS), while comparing differences between the two states. MethodsThirty female BABL/c mice were randomly divided into the normal group, the normal drug administration group, the CB group, the CB drug administration group and the dexamethasone group, with 6 mice in each group. The CB mouse model was established by inducing with ovalbumin (OVA). The mice in the normal drug administration group and the CB drug administration group started to be gavaged with XBS(13.2 g·kg-1) from the 21st day, and the dexamethasone group mice were simultaneously gavaged with dexamethasone (0.5 mg·kg-1) until the end of the 35th day of the experiment. Subsequently, serum samples were collected and evaluated for their efficacy, based on the pharmacological evaluation indicators, to determine the efficacy of XBS in treating CB. Then the UPLC-Q-Exactive Orbitrap MS was employed to identify and analyze the chemical constituents, blood-absorbed components, and metabolites of XBS. Chemometric analysis was conducted to reveal metabolic profile differences under "dual states". Concurrently, Real-time PCR technology was utilized to detect the expression levels of key liver metabolic enzymes CYP2E1, CYP3A1, UGT1A1, and UGT1A6. ResultsA total of 28 prototype components and 158 metabolites (including 48 phase Ⅰ metabolites and 110 phase Ⅱ metabolites) of XBS were unambiguously identified in the serum of normal mice. Additionally, a comprehensive characterization was performed on a total of 32 prototype components and 178 metabolites (including 50 phase Ⅰ metabolites and 128 phase Ⅱ metabolites) of XBS in the serum of CB mice. Among them, 27 prototype components were detected in both states, including 12 flavonoids, 2 alkaloids, 3 triterpenes, 4 organic acids, 3 amides, 1 stilbene and 2 other compounds. The chemometrics analysis revealed no significant difference in the prototype components and metabolites of XBS between normal and CB mice; however, there was a significant increase in the in-vivo exposure of XBS in CB mice. Compared to normal mice, the levels of phase Ⅰ metabolites such as oxidation, reduction and methylation of blood components of XBS as well as phase Ⅱ metabolites of glucuronidation showed significant changes in CB mice. Real-time PCR further confirmed that these alterations were attributed to the upregulation of CYP2E1 (P<0.05), CYP3A1 (P>0.05), UGT1A1 (P<0.01) and UGT1A6 (P<0.01) enzymes expression in the liver of CB mice. ConclusionThis study elucidated the disparities in the levels of the blood-absorbed components and metabolic profiles of XBS in normal and CB mice, especially in oxidation, reduction, methylation in phase Ⅰ metabolism and glucoaldehyde acidification in phase Ⅱ metabolism. And there are related to the differences in the expression levels of phase Ⅰ and phase Ⅱ metabolic enzymes CYP2E1, CYP3A1, UGT1A1 and UGT1A6 in the liver.
3.Study on preparation of universal platelet-rich plasma
Lu WANG ; Qiong WU ; Xinyue YAO ; Pingping MAO ; Yi YANG ; Kaiyun LUO ; Na LI ; Shujun WANG
Chinese Journal of Blood Transfusion 2026;39(7):840-845
Objective: To improve the efficiency of patient treatment in routine, emergency, and mass casualty scenarios (both peacetime and wartime), eliminate reliance on blood type compatibility, and promote the productization of platelet-rich plasma (PRP), this study investigated the optimal mixing ratio for developing a universal PRP formulation. Methods: PRP was prepared from donors with blood types O, A, B, and AB. Samples were mixed in varying proportions, and the optimal ratio was determined based on the trend of anti-A and anti-B antibody titers in the mixed PRP. Concentrations of the growth factors PDGF-AA, PDGF-BB, EGF, TGF-β1, and VEGF were measured by enzyme-linked immunosorbent assay (ELISA) before and after mixing. Results: A mixing ratio of A∶B∶AB=6∶2.5∶24 resulted in no detectable agglutination with either A or B cells in the microcolumn gel system, indicating the absence of immunological reactivity and confirming universal compatibility. Importantly, the levels of all measured growth factors remained unchanged following mixing, suggesting preservation of biological activity. Conclusion: These findings suggest that universal PRP can be prepared by neutralizing blood type antibodies through antigen-antibody reactions.
4.The Overview and Future Trend of High-quality Clinical Randomized Controlled Trial of Acupuncture for Neurological Disorders
Jin YAO ; Qiongnan BAO ; Kexin WU ; Wanqi ZHONG ; Xinyue ZHANG ; Ziwen CHEN ; Zihan YIN ; Fanrong LIANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(4):973-981
Objective Neurological disorders,constitute the majority of acupuncture-treated ailments.By critically examining a comprehensive selection of high-quality clinical randomized controlled trials(RCTs)from both domestic and international sources,we aimed to uncover the prevailing hot research topic and potential future directions in acupuncture therapy for these conditions.Methods Four databases,including PubMed,Web of science,Cochrane Library and EMBASE were searched.The search period is set from the establishment of each database to November 30,2023.And the search content was RCTs of acupuncture for neurological diseases classified as Ⅰ and Ⅱ by the Chinese Academy of Sciences were extracted.Key aspects extracted included annual publication volume,journals,disease types,outcomes,conclusions,acupuncture methods and control groups.Results A total of 81 high-quality RCTs were identified,revealing a consistent annual publication trend of acupuncture for neurological disorders.Key journals contributing to this research were Cephalalgia,Clinical Rehabilitation and Clinical Journal Of Pain,etc.The studies predominantly focused on migraine and sequela of apoplexy with migraine days/frequency and Ashworth scale and its modifications as commonly used primary outcome indicators,respectively.Overall,there were 73 RCTs that concluded that acupuncture for neurologic disorders were effective,8 were ineffective.The acupuncture methods were diverse,encompassing acupuncture,individualized acupuncture and transcutaneous electrical nerve stimulation,etc.The control measures were also varied,featuring sham acupuncture,sham-acupoint and micro-needle,etc.Conclusion With the development of acupuncture RCTs,their international influence has expanded.Research has shifted from validation of effectiveness to exploration of mechanisms,and despite the expansion of the disease spectrum of acupuncture,acupuncture trials have yet to be regulated and standardized,and the assessment of acupuncture efficacy remains controversial and challenging for the discipline.It is recommended to promote multi-center cooperation and strengthen the training of trialists etc.to provide more high-quality clinical evidence.
5.The impact of different bolus application methods on chest wall skin dose after radical mastectomy for breast cancer
Zuohuai HU ; Jiandong FU ; Xiaofang LI ; Xinyue YAO ; Bin ZHAO ; Shu YAN ; Sisi HE
The Journal of Practical Medicine 2025;41(14):2138-2142
Objective To investigate the impact of different application methods of tissue compensators(bolus)on the skin dose delivered to the chest wall following radical mastectomy for breast cancer.Methods A retrospective analysis was conducted on 60 female patients who underwent radical mastectomy and required chest wall radiotherapy at the hospital between January 2023 and March 2025.The Pinnacle3 9.10 radiotherapy planning system(TPS)was utilized to design two VMAT dual semi-arc radiotherapy plans for each patient,with a prescribed target dose of 50 Gy delivered in 2 Gy fractions over 25 sessions.In Plan 1,a Bolus was applied and optimized during the first 15 fractions,and subsequently removed for the remaining 10 fractions without re-optimization.The sub-field configuration and dose weighting from the initial optimization were retained,and only dose recalculations were performed.The final treatment plan combined both the Bolus-included and Bolus-excluded phases.In contrast,Plan 2 involved the application and optimization of Bolus during the first 15 fractions,followed by its removal and re-optimization of the plan for the last 10 fractions.The two optimized plans were then combined for the overall treatment delivery.Data from the two plan groups were analyzed using a paired sample t-test with SPSS 29.0 software.Results There was a statistically significant difference(P<0.05)in skin Dmean,V52.5,and V55;heart Dmean,V5,V30,and V40;affected lung Dmean,V5,and V20;PRVcord Dmean and Dmax;healthy breast Dmean,V5,and V10;affected humeral head Dmean and V30;as well as PTV Dmean,V50,V55,D2%,D98%,CI,and MU.Moreover,the dose distribution on the target layer and the DVH curves showed marked differences.However,no statistically significant difference was observed in PTV HI(P=0.125).Conclusion The combination of the two optimized plans,consisting of 15 fractions with bolus and 10 fractions without bolus,more accurately reflects the dose distri-bution within the planned target area and organs at risk,thereby providing enhanced protection for the patient's chest wall skin.
6.The Overview and Future Trend of High-quality Clinical Randomized Controlled Trial of Acupuncture for Neurological Disorders
Jin YAO ; Qiongnan BAO ; Kexin WU ; Wanqi ZHONG ; Xinyue ZHANG ; Ziwen CHEN ; Zihan YIN ; Fanrong LIANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(4):973-981
Objective Neurological disorders,constitute the majority of acupuncture-treated ailments.By critically examining a comprehensive selection of high-quality clinical randomized controlled trials(RCTs)from both domestic and international sources,we aimed to uncover the prevailing hot research topic and potential future directions in acupuncture therapy for these conditions.Methods Four databases,including PubMed,Web of science,Cochrane Library and EMBASE were searched.The search period is set from the establishment of each database to November 30,2023.And the search content was RCTs of acupuncture for neurological diseases classified as Ⅰ and Ⅱ by the Chinese Academy of Sciences were extracted.Key aspects extracted included annual publication volume,journals,disease types,outcomes,conclusions,acupuncture methods and control groups.Results A total of 81 high-quality RCTs were identified,revealing a consistent annual publication trend of acupuncture for neurological disorders.Key journals contributing to this research were Cephalalgia,Clinical Rehabilitation and Clinical Journal Of Pain,etc.The studies predominantly focused on migraine and sequela of apoplexy with migraine days/frequency and Ashworth scale and its modifications as commonly used primary outcome indicators,respectively.Overall,there were 73 RCTs that concluded that acupuncture for neurologic disorders were effective,8 were ineffective.The acupuncture methods were diverse,encompassing acupuncture,individualized acupuncture and transcutaneous electrical nerve stimulation,etc.The control measures were also varied,featuring sham acupuncture,sham-acupoint and micro-needle,etc.Conclusion With the development of acupuncture RCTs,their international influence has expanded.Research has shifted from validation of effectiveness to exploration of mechanisms,and despite the expansion of the disease spectrum of acupuncture,acupuncture trials have yet to be regulated and standardized,and the assessment of acupuncture efficacy remains controversial and challenging for the discipline.It is recommended to promote multi-center cooperation and strengthen the training of trialists etc.to provide more high-quality clinical evidence.
7.Exogenous triggering with hCG/GnRHa improves outcomes of natural cycle IVF/ICSI in patients with diminished ovarian reserve: a propensity score matching and logistic regression analysis.
Xinyue CHANG ; Ningning YAO ; Yan ZHAO ; Yinfeng WANG ; Ancong WANG ; Huihui ZHANG ; Jing ZHANG
Journal of Southern Medical University 2025;45(7):1519-1526
OBJECTIVES:
To explore the effects of exogenous trigger (hCG/GnRHa) versus endogenous LH surge in natural cycle IVF/ICSI (NC-IVF/ICSI) for patients with diminished ovarian reserve (DOR).
METHODS:
A retrospective analysis was conducted on 1,118 NC-IVF/ICSI cycles from two reproductive centers between 2013 and 2024. Propensity score matching (PSM) and multivariate logistic regression were used to adjust for confounding factors. The trigger-day hormone threshold was determined using receiver operating characteristic (ROC) curve analysis. Outcome measures included oocyte retrieval rate, 2PN fertilization rate, clinical available embryo rate, high-quality embryo rate, fresh cycle clinical pregnancy rate (CPR), and live birth rate (LBR).
RESULTS:
After adjusting for confounders via PSM and logistic regression, the exogenous trigger group demonstrated significantly better outcomes across all the evaluated parameters (oocyte retrieval rate, 2PN fertilization rate, transferable embryo rate, high-quality embryo rate, fresh cycle CPR, and LBR) than the endogenous LH surge group (P<0.05). Age-stratified analysis revealed that for the entire cohort, exogenous triggering significantly increased the number of transferable embryos and high-quality embryos (P<0.001). In the 35-39 years old subgroup, exogenous triggering showed significant advantages in oocyte yield, high-quality embryo rate, CPR, and LBR (P<0.05) and resulted in the most pronounced improvement in LBR (OR=6.25, 95% CI: 1.34-29.23). ROC analysis established a decision-day LH threshold of 19.055 mIU/mL (AUC=0.945, specificity=93.3%) for precise stratification of the clinical pathways.
CONCLUSIONS
For DOR patients undergoing NC-IVF/ICSI, exogenous triggering comprehensively improves the treatment outcomes, particularly providing significant live birth benefits for women aged 35-40 years. An individualized protocol incorporating the LH threshold (19.055 mIU/mL) effectively enhances embryonic developmental potential and live birth rates.
Humans
;
Female
;
Ovarian Reserve
;
Pregnancy
;
Propensity Score
;
Retrospective Studies
;
Fertilization in Vitro
;
Sperm Injections, Intracytoplasmic
;
Chorionic Gonadotropin
;
Pregnancy Rate
;
Logistic Models
;
Ovulation Induction/methods*
;
Gonadotropin-Releasing Hormone
;
Adult
;
Oocyte Retrieval
8.Association of takeaway consumption and sedentary behavior with emotional symptoms among freshman students
YAO Zhiyuan, WEI Ruihong, WANG Xinyue, JIANG Linlin, WAN Yuhui
Chinese Journal of School Health 2025;46(10):1474-1477
Objective:
To investigate the assocation of sedentary behavior among college students on psychological health issues, such as depressive, anxiety, and stress symptoms, and to analyze the moderating role of takeaway consumption behavior in the context, in order to provide a scientific basis for reducing emotional symptoms among college students.
Methods:
A stratified cluster sampling method was employed to conduct a questionnaire on 3 427 first year students of a higher education institution in Hefei of Anhui Province from May to June 2021. The study variables included demographic characteristics, sedentary time, takeaway consumption behavior, and emotional (symptoms depressive, anxiety, and stress symptoms). The Spearman correlation analysis was used to analyze the association between variables, and linear regression analysis was used to analyze the association between takeaway consumption behavior and depressive, anxiety and stress symptoms among college students with sedentary time.
Results:
Both sedentary time and takeaway consumption behavior were positively correlated with depressive, anxiety and stress symptoms among college students ( r =0.10, 0.10, 0.10; 0.10, 0.11, 0.11, all P <0.05). The results of linear regression analysis showed that the interaction term between takeaway consumption behavior and sedentary time was positively correlated with symptoms of depressive, anxiety, and stress symptoms among college students (depression: β =0.04, anxiety: β =0.04, stress: β =0.04, all P <0.05). The results of the simple slope test demonstrated that regardless of the level of takeaway consumption behavior, sedentary time was positively correlated with the depressive symptoms of college students; compared with low takeaway consumption behavior, high takeaway consumption behavior ( β=0.77, P <0.01) enhanced the association between sedentary time and depressive symptoms among college students. In addition, under the condition of high takeaway consumption behavior, sedentary time was positively correlated with the anxiety and stress symptoms of college students (anxiety: β =0.64; stress: β =0.71, both P <0.01); while under the condition of low takeaway consumption behavior, sedentary time was not related to the anxiety and stress symptoms of college students ( β =0.17, 0.22, both P >0.05).
Conclusions
Sedentary behavior is related to a the emotional symptoms of depressive, anxiety, and stress among college students. Takeaway consumption behavior may exacerbate this impact.
9.Study on the role of butyric acid-producing bacteria in periodontitis-induced interference with long bone homeostasis in mice fed a high-fat/high-sugar diet
XU Zhonghan ; YAO Yujie ; WANG Xinyue ; SONG Shiyuan ; BAO Jun ; YAN Fuhua ; TONG Xin ; LI Lili
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(6):445-456
Objective:
To investigate the role of butyric acid-producing bacteria in long bone homeostasis in mice with periodontitis under a high-fat/high-sugar diet and to provide new insights for the prevention and treatment of periodontitis and related bone metabolic diseases.
Methods:
This study has been approved by the Animal Welfare and Ethics Committee of the Experimental Animal Center. Initially, 14 mice were randomly divided into the CON group (the control group) and the LIG group (the periodontitis group). Mice in the LIG group had experimental periodontitis induced by ligating the second maxillary molars bilaterally and were fed a high-fat and high-sugar diet. After 8 weeks, samples were collected. Micro-computed tomography (Micro-CT) was used to analyze alveolar bone resorption and various parameters of the proximal tibia trabecular bone, including bone mineral density (BMD), bone volume per tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp). After decalcification, hematoxylin and eosin (HE) staining was performed on maxillary bone sections to assess periodontal tissue inflammation and connective tissue destruction. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect related genes in the distal femur and proximal tibia bone tissues, including osteocalcin (OCN), osteogenic transcription factor (Osterix), osteoprotegerin (OPG), tartrate resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), receptor activator of nuclear factor kappa-B (RANK), and receptor activator of nuclear factor kappa-B ligand (RANK-L). Subsequently, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + butyric acid-producing bacteria (BP) group, and LIG + BP group. The breeding, sampling, and sample detection methods remained the same. Finally, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + sodium butyrate (SB) group, and LIG + SB group. The breeding, sampling, and sample detection methods remained the same.
Results:
①Periodontitis modeling was successful. Compared with the CON group, the LIG group exhibited significant alveolar bone resorption of the maxillary second molar, aggravated periodontal tissue inflammation, and connective tissue destruction. ②Periodontitis exacerbated long bone resorption in mice fed a high-fat high-sugar diet. Compared with the CON group, the LIG group had significantly lower BMD, BV/TV, Tb.N, and Tb.Th (P<0.05), and significantly higher Tb.Sp (P<0.05). HE staining of the proximal tibia showed that the trabeculae in the LIG group were sparse and disordered, with some areas showing fractures or dissolution. The expression of osteoblast markers (OCN, Osterix, OPG) was significantly lower in the LIG group (P<0.05), while the expression of the osteoclast marker TRAP showed an increasing trend (P>0.05). The ratio of RANK-L/OPG was significantly higher in the LIG group compared with the CON group (P<0.05). ③ Supplementation with butyric acid-producing bacteria alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BMD and Tb.Th were significantly higher in the LIG + BP group. HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + BP group compared with the LIG group. The expression of OCN and Osterix was significantly higher in the LIG + BP group, while the expression of osteoclast-specific genes (OSCAR, RANK, RANK-L) was significantly lower (P<0.05). ④ Supplementation with butyrate alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BV/TV and Tb.N were significantly higher in the LIG + SB group, and Tb.Sp was significantly lower (P<0.05). HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + SB group compared with the LIG group. The expression of Osterix, OPG, OSCAR, TRAP, and RANK was significantly lower in the LIG + SB group compared with the LIG group (P<0.05).
Conclusion
Periodontitis disrupts the long bone homeostasis of mice fed a high-fat high-sugar diet, aggravating long bone resorption. Supplementation with butyric acid-producing bacteria or butyrate can effectively alleviate the disruption of long bone homeostasis caused by periodontitis.
10.Elesclomol combined with Cu2 + induced cuproptosis in hepatoma cell lines PLC/PRF/5 and Huh-7
Qiaohui Ren ; Xinyue Zhu ; Wei Lv ; Yan Zang ; Lianzi Wang ; Xinyi Zhou ; Junxiao Yao ; Tao Li
Acta Universitatis Medicinalis Anhui 2025;60(8):1470-1477
Objective:
To investigate the inhibitory effect of Elesclomol (ES) + Cu2 + on the proliferation of human hepatoma cell lines PLC/PRF/5 and Huh-7 and its potential to induce Cuproptosis.
Methods:
Human hepatoma cell lines PLC/PRF/5 and Huh_7 cells were Cultured in vitro. ES solution , Cu2 + solution and copper chelating agent ammonium tetrathiomolybdate VI (ATTM) solution was treated separately or in combination. The effect of ES + Cu2 + on the survival rate of human hepatoma cell lines PLC/PRF/5 and Huh_7 cells and the effect of ES + Cu2 + on the survival rate after pretreatment with copper chelating agent ATTM were evaluated using CCK_8 kit. The cell death induced by ES + Cu2 + was detected by flow cytometry and the changes of ES + Cu2 + after pretreatment with copper chelating agent ATTM. The expression of Cuproptosis related proteins ATPase copper transporting beta (ATP7B) ,ferredoxin 1 (FDX1) , dihydrolipoamide s_acetyltransferase(DLAT) and superoxide dismutase 1 (SOD1) were detected by Western blot. The effect of ES + Cu2 + on cell proliferation and the reverse effect after ATTM pretreatment was detected by cell scratch assay.
Results:
The toxicity of ES + Cu2 + to human hepatocellular carcinoma cell lines PLC/PRF/5 and Huh_7 was significantly dose_dependent (P < 0. 05) . Compared with the control group , the combined application of ES and Cu2 + had a more significant inhibitory effect on hepatocellular carcinoma cells than ES or Cu2 + alone (P < 0. 05) , and copper chelating agent ATTM could reverse the inhibitory effect of ES + Cu2 + on hepatocellular carcinoma cells (P < 0. 05) . Flow cytometry results showed that compared with the control group , the proportion of cell death in PLC/PRF/5 and Huh_7 cells treated with ES + Cu2 + increased , while the proportion of cell death decreased after ATTM intervention (P < 0. 05) . The results of cell scratch test showed that the migration ability of PLC/PRF/5 and Huh_7 cells was decreased after ES + Cu2 + treatment , however, the addition of ATTM reversed the inhibitory effect of ES + Cu2 + on cell migration (P < 0. 05) . Compared with the control group , the expression levels of copper death related proteins ATP7B , FDX1 , DLAT and SOD1 decreased after ES + Cu2 + treatment , but the addition of ATTM reversed the expression trend of these proteins (P < 0. 05) .
Conclusion
The combination of ES and Cu2 + can effectively inhibit the proliferation and migration of PLC/PRF/5 and Huh_7 of hepatocellular carcinoma cells , and induce Cuproptosis , which provides a new strategy for the treatment of hepatocellular carcinoma.


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