1.Mechanism of Modified Si Junzitang and Shashen Maidong Tang in Improving Sensitivity of Cisplatin in EGFR-TKI Resistant Lung Adenocarcinoma Cells Based on Aerobic Glycolysis
Yanping WEN ; Yi JIANG ; Liping SHEN ; Haiwei XIAO ; Xiaofeng YANG ; Surui YUAN ; Lingshuang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):39-46
ObjectiveTo investigate the mechanism of modified Si Junzitang and Shashen Maidong Tang [Yiqi Yangyin Jiedu prescription (YQYYJD)] in enhancing the sensitivity of cisplatin in epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI)-resistant lung adenocarcinoma cells based on aerobic glycolysis. MethodsThe effects of different concentrations of YQYYJD (0, 2, 3, 4, 5, 6, 7, 8 g·L-1) and cisplatin (0, 3, 6, 9, 12, 15, 18, 21, 24, 27 mg·L-1) on the proliferation and activity of PC9/GR cells were detected by the cell counting kit-8 (CCK-8) assay after 24 hours of intervention. The half-maximal inhibitory concentration (IC50) for PC9/GR cells was calculated to determine the concentrations used in subsequent experiments. PC9/GR cells were divided into blank group (complete medium), YQYYJD group (5 g·L-1), cisplatin group (12 mg·L-1), and combined group (YQYYJD 5 g·L-1 + cisplatin 12 mg·L-1). After 24 hours of intervention, cell viability was measured using CCK-8 assay. Cell proliferation was assessed by colony formation assay, and cell migration was evaluated by scratch and Transwell assays. Glucose consumption, lactate production, and adenosine triphosphate (ATP) levels were measured by colorimetric assays. The expression levels of glycolysis-related proteins, including hexokinase 2 (HK2), phosphofructokinase P (PFKP), pyruvate kinase M2 (PKM2), lactate dehydrogenase A (LDHA), glucose transporter 1 (GLUT1), and monocarboxylate transporter 4 (MCT4), were determined by Western blot. ResultsBoth YQYYJD and cisplatin inhibited the viability of PC9/GR cells in a concentration-dependent manner. The IC50 of PC9/GR cells for YQYYJD and cisplatin were 5.15 g·L-1 and 12.91 mg·L-1, respectively. In terms of cell proliferation, compared with the blank group, the cell survival rate and the number of colonies formed in the YQYYJD group, cisplatin group, and combined group were significantly decreased (P<0.01). Compared with the YQYYJD and cisplatin groups, the combined group showed a further significant reduction in cell survival rate and colony formation (P<0.01). In terms of cell migration, compared with the blank group, the cell migration rate and the number of cells passing through the Transwell membrane in the YQYYJD group, cisplatin group, and combined group were significantly decreased (P<0.01). Compared with the YQYYJD and cisplatin groups, the combined group exhibited a further significant reduction in cell migration rate and the number of cells passing through the Transwell membrane (P<0.01). In terms of glycolysis, compared with the blank group, glucose consumption, lactate production, and ATP levels in the YQYYJD group, cisplatin group, and combined group were significantly decreased (P<0.01). Compared with the YQYYJD and cisplatin groups, the combined group showed a further significant reduction in glucose consumption, lactate production, and ATP levels (P<0.05). Compared with the blank group, the protein expression levels of HK2, PFKP, PKM2, and LDHA in the YQYYJD, cisplatin, and combined groups were significantly decreased (P<0.01). The combined group showed a further significant reduction in the expression levels of these proteins compared with the YQYYJD and cisplatin groups (P<0.01). No significant differences were observed in the protein expression levels of GLUT1 and MCT4 among the groups. ConclusionYQYYJD can synergistically inhibit the proliferation and migration of PC9/GR cells and enhance their sensitivity to cisplatin. The mechanism may be related to the downregulation of the expression of glycolysis-related rate-limiting enzymes, including HK2, PFKP, PKM2, and LDHA, thereby inhibiting glycolysis.
2.Effect of Yiqi Yangyin Jiedu Prescription on sPD-L1 in Peripheral Blood of Patients Treated with EGFR-TKIs and Prognosis Analysis
Liping SHEN ; Yuqing CAI ; Yanping WEN ; Yi JIANG ; Lingshuang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):219-226
ObjectiveTo observe the prognosis effect of soluble programmed death ligand-1(sPD-L1) in treating patients with advanced lung adenocarcinoma treated with epidermal growth factor receptor-tyrosine kinase inhibitors(EGFR-TKIs) and the influence of Yiqi Yangyin Jiedu prescription. MethodA prospective cohort-controlled study was conducted to enroll patients treated with EGFR-TKIs in the first line of treatment,who were admitted to the Oncology Department of Longhua Hospital and Shanghai Chest Hospital from May 1st, 2021 to June 30th, 2023, and they were evaluated as non-progressive and identified with deficiency of Qi and Yin after one month of treatment. The patients were divided into an exposed group (EGFR-TKIs combined with Yiqi Yangyin Jiedu prescription) and a non-exposed group (EGFR-TKIs alone)according to whether or not they were treated with Yiqi Yangyin Jiedu prescription and were treated until disease progression, or death and intolerable adverse reactions occurred. The enzyme-linked immunosorbent assay (ELISA) was applied to detect the level of sPD-L1 in patients at the time of enrollment and disease progression,and Cox risk proportionality model was used to analyze the independent prognostic factors affecting disease progression of patients treated with EGFR-TKIs. ResultA total of 90 patients (39 in the exposed group and 51 in the non-exposed group) undergoing disease progression after EGFR-TKI treatment were enrolled. At the time of enrolment and after disease progression,the levels of serum sPD-L1 in the 90 patients were 12.06 (27.54) ng·L-1 and 41.99 (62.93) ng·L-1,respectively. Compared with that at the time of enrollment, the serum sPD-L1 level in the 90 patients was significantly increased after disease progression (P<0.01). The serum sPD-L1 level in patients in the exposed group was 12.27 (24.78) ng·L-1 and 29.57 (61.12)ng·L-1 respectively at the time of enrolment and after disease progression. In the non-exposed group, patients had serum sPD-L1 levels of 11.81 (28.46) ng·L-1 and 49.54 (74.12) ng·L-1 respectively at the time of enrolment and after disease progression. Compared with that at the time of enrollment, the serum sPD-L1 level in the two groups of patients was significantly increased after disease progression (P<0.01). In addition, compared with that in the non-exposed group, the sPD-L1 level in the exposed group was greatly reduced after disease progression(P<0.01). Cox multifactorial analysis showed that sPD-L1 level and age at the time of enrolment were associated with patients' progression-free survival(PFS),and that low levels of sPD-L1 (<12.06 ng·L-1) prolonged the PFS and reduced the risk of disease progression in patients treated with EGFR-TKIs compared with high levels of sPD-L1. ConclusionElevated sPD-L1 level is a poor prognostic factor for the long-term efficacy of EGFR-TKIs,and treatment with Yiqi Yangiin Jiedu prescription can down-regulate sPD-L1 level of patients treated with EGFR-TKIs.
3.Efficacy Mechanism of Xianlian Jiedu Prescription Against Colorectal Cancer Recurrence vias Regulating Angiogenesis
Yanru XU ; Lihuiping TAO ; Jingyang QIAN ; Weixing SHEN ; Jiani TAN ; Chengtao YU ; Minmin FAN ; Changliang XU ; Yueyang LAI ; Liu LI ; Dongdong SUN ; Haibo CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):79-87
ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer (CRC) and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit, followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group, low-dose Xianlian Jiedu prescription (XLJDP-L) group (6.45 g·kg-1·d-1), medium-dose Xianlian Jiedu prescription (XLJDP-M) group (12.9 g·kg-1·d-1), high-dose Xianlian Jiedu prescription (XLJDP-H) group (25.8 g·kg-1·d-1), and 5-fluorouracil (5-FU) group (1×10-3 g·kg-1·d-1). The mice were euthanized after 14 days of continuous intervention, and recurrent tumor tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry (IHC) was employed to assess the expression of proliferating cell nuclear antigen (Ki67), vascular endothelial growth factor (VEGF), and platelet-endothelial cell adhesion molecule (CD31) in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 (ANG-2), VEGF, phosphorylated-protein kinase B (p-Akt), protein kinase B (Akt), phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), and phosphatidylinositol 3-kinase (PI3K) in recurrent tumor tissue. ResultsBefore treatment, there were no statistical differences in tumor volume, tumor weight, and body mass among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group, indicating model stability. After treatment, compared with those in the model group, the tumor volume and tumor weight in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01), showing dose dependency. Meanwhile, there were no significant differences in body weight among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group, tumor tissue in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group had loosely arranged cells, increased intercellular spaces, small and shriveled nuclei, light staining, fewer mitotic figures and atypical nuclei, and increased necrotic areas. IHC showed that compared with those of the model group, the positive rates of Ki67, VEGF, and CD31 in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01) in a dose-dependent manner. Western blot results showed that compared with those of the model group, the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly downregulated (P<0.05, P<0.01), and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner (P<0.05, P<0.01). ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2, VEGF, and CD31.
4.Study on the correlation between inflammatory indicators in the hyperacute phase of acute ischemic stroke and early neurological deterioration, syndrome factors, and prognosis
Ligaoge KANG ; Ying GAO ; Jinyue BAI ; Huan TANG ; Hongbo SHEN ; Lei LIU ; Lingbo KONG
Journal of Beijing University of Traditional Chinese Medicine 2025;48(1):98-107
Objective:
Inflammatory cascade reactions play a crucial role in secondary neuronal injury in acute ischemic stroke (AIS). The aim of this study was to explore the correlations between specific serological indicators, early neurological deterioration (END), disease prognosis, and syndrome factors in AIS based on this injury mechanism.
Methods:
The data for this study were collected from 135 patients with AIS admitted to the emergency department of Fangshan Hospital, Beijing University of Chinese Medicine, within 24 h of onset between November 2019 and May 2021. Among these, 29 patients had complete data and experienced END. Additionally, 9 non-END patients were matched from the remaining 90 patients with complete data, resulting in a total of 38 patients for statistical analysis. Statistical methods, including logistic regression and receiver operating curves, were used to analyze the correlation between serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), vascular endothelial growth factor (VEGF), and intercellular adhesion molecule-1 (ICAM-1) within 24 h of END onset, disease prognosis, and syndrome factors. Grouping criteria included END occurrence, presence of syndrome elements on the first and third day post-onset, and prognosis at 90 days post-onset.
Results:
All 38 cases had onset time of less than 12 h, and there were no significant differences in age, gender, and onset time between the END and non-END groups. The TNF-α serum level within 24 h of onset was not associated with the occurrence of END but was negatively correlated with all-cause mortality at 90 days [0.1
5.Relationship between levels of novel inflammatory indicators and aggressivity in patients with first-episode and recurrent schizophrenia
Ying'ao CUI ; Cheng YANG ; Yinghan TIAN ; Qingqing SHEN ; Huanzhong LIU
Sichuan Mental Health 2025;38(1):28-33
BackgroundAggressive behavior in schizophrenic patients could result in legal disputes and public safety concerns. In patients with illness episodes of different numbers, there may exist differences in the association between levels of novel inflammatory indicators and aggressivity. ObjectiveTo investigate the differences in the correlation between levels of novel inflammatory indicators and aggressivity in patients with first-episode and recurrent schizophrenia, in order to search for inflammatory biomarkers to assess aggression level in schizophrenic patients. MethodsA total of 168 schizophrenic patients were selected as subjects, who were hospitalized for acute disease onset in Chaohu Hospital of Anhui Medical University from October 2022 to April 2024 as well as met the diagnostic criteria of Diagnostic and Statistical Manual of Mental Disorders, fifth edition (DSM-5). Patients were divided into first-episode group (n=58) and recurrent group (n=110). Meanwhile, 110 healthy controls from community who matched in age and gender with the patient group were recruited. All patients were evaluated with Modified Overt Aggression Scale (MOAS) and Positive and Negative Syndrome Scale (PANSS). All subjects went through examination of the levels of novel inflammatory indicators, including neutrophil/lymphocyte ratio (NLR), monocyte/lymphocyte ratio (MLR), platelet/lymphocyte ratio (PLR), neutrophil/high-density lipoprotein ratio (NHR), monocyte/high-density lipoprotein ratio (MHR) and platelet/high-density lipoprotein ratio (PHR). Spearman correlation analysis was adopted to investigate the correlation between levels of novel inflammatory indicators and the total score of MOAS in patients with first-episode and recurrent schizophrenia. ResultsThe levels of NLR, MLR, PLR, NHR, MHR and PHR in first-episode group were higher than those in control group (adjusted P<0.01). The levels of NLR, MLR, NHR, MHR and PHR in recurrent group were higher than those in control group (adjusted P<0.01). No significant difference was observed in the comparison in the levels of six novel inflammatory indicators between first-episode group and recurrent group (adjusted P>0.05). Spearman correlation analysis showed, the MOAS total score of recurrent group was positively correlated with the levels of NLR, MLR and PLR (r=0.234, 0.192, 0.243, P<0.05). There was no statistical significance in the correlation between MOAS total score and levels of six novel inflammatory indicators in first-episode group (P>0.05). ConclusionAmong patients with first-episode and recurrent schizophrenia, the correlation between levels of novel inflammatory indicators and aggressivity could differ. NLR, MLR and PLR might be the biomarkers for assessing aggression level in recurrent schizophrenic patients. [Funded by Anhui Provincial Natural Science Foundation (number, 2108085MH275)]
6.The renoprotective effect of Salvia miltiorrhiza injection on HBOC-CHP01 resuscitated haemorrhagic shock rats
Shasha HAO ; Xintong XIE ; Shen LI ; Honghui ZHANG ; Hong WANG ; Chengmin YANG ; Wentao ZHOU ; Jiaxin LIU
Chinese Journal of Blood Transfusion 2025;38(2):172-180
[Objective] To investigate the protective effect of Salvia miltiorrhiza injection (SMI) on the kidneys of HBOC-CHP01 resuscitated haemorrhagic shock rats. [Methods] A 50% haemorrhagic shock rat model was established, with 12 rats divided into two groups: SMI + HBOC-CHP01 group and HBOC-CHP01 group, with 6 rats in each group. The rats in the SMI+ HBOC-CHP01 group were given an equal volume of HBOC-CHP01 for resuscitation after haemorrhagic shock, and an 8 mL/kg dose of SMI. Rats in the HBOC-CHP01 group were resuscitated by administering an equilibrium blood loss volume of HBOC-CHP01 and given an 8 mL/kg dose of 0.9% NaCl solution. Blood was taken from rats at five points: before bloodletting (baseline), during haemorrhagic shock (HS), immediately after resuscitation (RS0h), 1 h after resuscitation (RS1h), and 24 h after resuscitation (RS24h). A blood gas analyser was used to detect the lactate level (Lac), glucose content (Glu), residual base (BEecf), pH, bicarbonate (HCO3-), high iron haemoglobin (MetHb). White blood cells (WBC), platelets (PLT), haemoglobin content (Hb), carboxyhaemoglobin (COHb) were detected using a quintuple classification. Blood creatinine (SCr), uric acid (UA), kidney-related indexes were detected using biochemistry instrument. Kidney tissues of the rats were taken after 24 h of resuscitation and after execution, and the inflammation of kidneys of the rats of the two groups was analyzed using HE staining. Fluorescence staining was used to detect the level of ROS in the kidneys of rats in both groups. [Results] At RS 0h, the Beecf, Glu and Lac levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group, and the pH level of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group, and the Glu levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group at RS 1h. At RS 0h, the WBC, PLT and COHb contents of rats in the SMI+HBOC-CHP01 group were all significantly higher than those of rats in the HBOC-CHP01 group, and at RS 1h, the WBC content of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group; at RS 1h, the UA content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the SCr content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the inflammation level of kidney tissues of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC -CHP01 group rats, and the ROS and MPO levels in the kidney tissues of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group. [Conclusion] The combination of Salvia miltiorrhiza injection during the resuscitation of rats with severe haemorrhagic shock by HBOC-CHP01 can alleviate renal injury by reducing inflammatory response and oxidative stress.
7.The multi-center mid-term clinical outcomes of combined complete preservation of chordal structure mitral valve replacement with total anatomic complete arterial myocardial revascularization for coronary patients with moderate-to-severe or severe ischemic mitral regurgitation
Ke GUO ; Xujun CHEN ; Baoshi ZHENG ; Chao SHI ; Keli HUANG ; Yong CAO ; Chengquan LIAO ; Jingwei CHEN ; Yu LIN ; Chengxin LIU ; Quansheng CAO ; Lin SHEN ; Zhendong WANG
Chinese Journal of Surgery 2025;63(1):58-67
Objective:To evaluate the clinical outcomes of combined complete preservation of chordal structure mitral valve replacement (C-MVR) with total anatomical arterial myocardial revascularization (TACR) in coronary patients with moderate-to-severe or severe ischemic mitral regurgitation (IMR).Methods:This is a retrospective multi-center case series study. Data were retrospectively collected from 127 patients with coronary artery disease with moderate to severe or severe IMR who received TACR with C-MVR from July 2015 to April 2024 in 13 hospitals in China. There were 90 males and 37 females, aged (56.5±10.7) years (range: 33 to 74 years). Perioperative data and follow-up data including left ventricular ejection fraction, left ventricular end-diastolic diameter, and patency rate of arterial grafts of patients were collected. Comparisons were made using paired sample t-test or χ2 test. Results:In this cohort of 127 patients, 67 underwent concurrent tricuspid valve repair. During surgery, 113 grafts of the left internal mammary artery (LIMA), 127 grafts of the left radial artery, 80 grafts of the right radial artery, and 110 grafts of the right internal mammary artery (RIMA) were harvested. The number of the distal anastomosis was 4.2±0.4 (range: 3 to 5). The aortic cross-clamp time and cardiopulmonary bypass time were (97.5±23.4) minutes (range: 90 to 161 minutes) and (145.4±19.2) minutes (range: 101 to 210 minutes), respectively. There was one operative death. Intraoperative placement of an intra-aortic balloon pump was performed in 21 patients to improve the left ventricular ejection. No sternal ischemic occurred. All patients completed follow-up, with a mean follow-up period of (64.3±7.5) months (range: 4 to 110 months). No major cerebrovascular events occurred during the follow-up period, and all patients survived. Left ventricular ejection fraction improved postoperatively (55.0%±5.3% vs. 41.0%±15.3%, t=17.23, P<0.01). The proportion of patients with New York Heart Association functional class ≤2 increased postoperatively (23.6% (30/127) vs. 87.3% (110/126), χ2=103.77, P<0.01). The proportion of patients with Canadian Cardiovascular Society Angina Classification ≤3 decreased postoperatively (4.8% (6/126) vs. 78.7% (100/127), χ2=142.19, P<0.01). The left ventricular end-diastolic diameter decreased postoperatively ((5.70±4.50) cm vs. (6.10±0.23) cm, t=12.15, P<0.01). Coronary multi-detector computed tomography angiography (MDCTA) follow-up was conducted for (60.5±11.7) months (range: 6 to 109 months) postoperatively. MDCTA confirmed the patency rates of the grafts: 96.4% (108/112) for the LIMA grafts, 88.9% (112/126) for the left radial artery grafts, 93.7% (74/79) for the right radial artery grafts, and 90.9% (100/110) for the free RIMA grafts. No significant differences in graft patency rates were observed between the arterial grafts ( χ2=5.24, P=0.155). Conclusion:The results of this multi-centre study demonstrate satisfactory mid-term results of C-MVR with TACR for the treatment of coronary artery disease with moderate to severe or severe IMR.
8.Clinical diagnosis of drug-induced autoimmune-like hepatitis
Journal of Clinical Hepatology 2025;41(3):542-546
Some patients with drug-induced liver injury (DILI) may present with similar features to patients with autoimmune hepatitis (AIH), and such cases are defined as drug-induced autoimmune-like hepatitis (DI-ALH). In 2022, the European Association for the Study of the Liver discussed the nomenclature, diagnosis, and treatment of DI-ALH and put forward expert opinions to establish the criteria for clinical diagnosis. However, the clinical diagnosis of DI-ALH remains complex and difficult, and there is no direct evidence for the link between drug intake and the body’s immune progression. Therefore, this article reviews the research advances in the diagnosis of DI-ALH from the four aspects of clinical phenotype, pathological diagnosis, diagnostic score, and clinical prognosis.
9.Mechanism of action of bile-gut axis in the development and progression of intrahepatic cholangiocarcinoma
Xue YU ; Tianhao SHEN ; Cheng ZHOU ; Yu LIU ; Wei LI ; Tinghui JIANG ; Yongqiang ZHU ; Yan LIU
Journal of Clinical Hepatology 2025;41(3):588-593
Intrahepatic cholangiocarcinoma is a malignant tumor with an extremely poor prognosis, and its pathogenesis is complex and remains unclear. In recent years, more and more studies have focused on the role of bile-gut axis in the development and progression of intrahepatic cholangiocarcinoma. Bile-gut axis refers to the complex interaction between bile and gut microbiota, including bile salt metabolism, dynamic changes of microbiota, inflammatory response, and immune system regulation. This article elaborates on the potential mechanisms of bile-gut axis in intrahepatic cholangiocarcinoma, especially gut microbiota dysbiosis, abnormal bile salt metabolism, chronic inflammatory response, and immune system interaction, this article aims to provide new perspectives and possible therapeutic targets for future research and promote the early diagnosis and effective treatment of intrahepatic cholangiocarcinoma.
10.Analysis of Mechanism of Astragaloside Ⅳ in Regulating Ferroptosis Through SLC7A11/GPX4 Pathway Against Vascular Smooth Muscle Cell Proliferation
Guoting LI ; Changchao YANG ; Lin LIU ; Weikang LI ; Zixian ZHAO ; Quan SHEN ; Jingshan ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):159-167
ObjectiveTo investigate the effect of astragaloside Ⅳ(AS-Ⅳ) on the proliferation of vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ(Ang Ⅱ) based on solute carrier family 7 member 11/glutathione peroxidase 4(SLC7A11/GPX4) pathway. MethodsPrimary rat thoracic aortic VSMCs were cultured by tissue explant method, and the cell types were identified by immunofluorescence. Cell counting kit-8(CCK-8) was used to determine the optimal concentration and time of AS-Ⅳ after Ang Ⅱ stimulation. The experiment was divided into blank group, model group, AS-Ⅳ group(40 μmol·L-1), Erastin group(0.5 μmol·L-1), Erastin+AS-Ⅳ group(0.5 μmol·L-1+40 μmol·L-1). The blank group was cultured in normal medium, the model group was cultured in medium containing Ang Ⅱ(0.1 μmol·L-1), and each administration group was cultured in medium containing Ang Ⅱ(0.1 μmol·L-1) and the corresponding doses of drug. CCK-8 and plate clone formation assay were used to detect the proliferation of cells in each group, Prussian blue staining was used to detect cell iron deposition, the content of reactive oxygen species(ROS) in cells was detected by fluorescence probe method, the content of malondialdehyde(MDA) was detected by thiobarbituric acid(TBA) method, and the protein levels of SLC7A11 and GPX4 in each group were detected by Western blot. ResultsPrimary rat thoracic aortic VSMCs were successfully cultured by tissue explant method, and immunofluorescence detection showed that positive expression of α-smooth muscle actin(α-SMA) and negative expression of vimentin in the cells, identifying them as VSMCs. The optimal concentration and time of AS-Ⅳ determined by CCK-8 were 40 μmol·L-1 and 24 h, respectively. Pharmacodynamic studies showed that compared with the blank group, the cell proliferation in the model group increased, the iron deposition in the cells increased, the contents of ROS and MDA increased, and the expression levels of SLC7A11 and GPX4 proteins decreased(P<0.05, P<0.01). Compared with the model group, the cell proliferation of the AS-Ⅳ group was inhibited, the iron deposition in the cells was decreased, the contents of ROS and MDA were decreased, and the expression levels of SLC7A11 and GPX4 proteins were increased(P<0.05, P<0.01). While in the Erastin group, the cell proliferation was increased, the iron deposition was increased, ROS and MDA contents were increased, and the expression levels of SLC7A11 and GPX4 proteins were decreased(P<0.05, P<0.01). Compared with the AS-Ⅳ group, Erastin+AS-Ⅳ group showed increased cell proliferation, increased iron deposition in cells, increased ROS and MDA contents, and decreased expression of SLC7A11 and GPX4 proteins(P<0.05). Compared with the Erastin group, the cell proliferation in Erastin+AS-Ⅳ group was inhibited, the iron deposition was decreased, the contents of ROS and MDA were decreased, and the expression levels of SLC7A11 and GPX4 proteins were increased(P<0.05, P<0.01). ConclusionAS-Ⅳ can inhibit ferroptosis by regulating the SLC7A11/GPX4 pathway, so as to weaken the proliferation of VSMCs, thus playing a role in the treatment of atherosclerosis.


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