1.Effects of thioredoxin reductase 1 on ferroptosis and immune function of dendritic cells in septic mice
Qiyuan ZHOU ; Jingyan LI ; Yanmin CAO ; Weiling LI ; Ning DONG ; Yao WU ; Yingping TIAN ; Yongming YAO
Chinese Journal of Burns 2025;41(3):212-221
Objective:To investigate the effects of thioredoxin reductase 1 (TXNRD1) on ferroptosis and immune function of dendritic cells (DCs) in septic mice, and to provide a basis for improving the immunosuppression in sepsis caused by wound infection.Methods:This study was an experimental research. Sixty male C57BL/6J mice aged 6-8 weeks were subjected to cecal ligation and puncture (CLP) to establish sepsis models. Ten mice were selected at 0 (immediately), 6, 12, 24, 48, and 72 h after CLP surgery, respectively, according to the random number table method. Mouse splenic DCs were isolated using CD11c-positive magnetic beads. The protein expressions of TXNRD1, and anti-ferroptosis proteins solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the cells were detected by Western blotting, the reduced glutathione (GSH) content in the cells was measured by colorimetric assay, the lipid peroxidation level was assessed via live-cell imaging technology, and the levels of major histocompatibility complex class Ⅱ subtype I-A/I-E and leukocyte differentiation antigens CD80 and CD86 were detected by flow cytometry. Another 100 male C57BL/6J mice aged 6-8 weeks were divided into corn oil+sham injury group, corn oil+CLP group, inhibitor+sham injury group, and inhibitor+CLP group according to the random number table method, with 25 mice in each group. Mice in the two inhibitor groups were intraperitoneally injected with TXNRD1 inhibitor auranofin, while mice in the two corn oil groups were intraperitoneally injected with corn oil. One hour later, mice in the two CLP groups underwent CLP surgery to establish sepsis models, while mice in the two sham injury groups underwent sham surgery. Twenty mice from each group were selected to observe survival within 7 d post-surgery, and the survival rate was calculated. At 24 h post-surgery, mouse splenic DCs from the remaining 5 mice in each group were collected for corresponding assays as above.Results:Compared with those at 0 h after CLP surgery, the protein expressions of TXNRD1, GPX4, and SLC7A11 in mouse cells at 24 h after CLP surgery and the protein expression of TXNRD1 in mouse cells at 48 h after CLP surgery were significantly decreased ( P<0.05), the GSH content in mouse cells was significantly decreased at 24 and 48 h after CLP surgery ( P<0.05). The lipid peroxidation level in mouse cells was low at 0, 6, and 12 h after CLP surgery, slightly lower than that at 72 h after CLP surgery; the lipid peroxidation levels in mouse cells at 24 and 48 h after CLP surgery were significantly higher than those at 0, 6, 12, and 72 h after CLP surgery. Compared with those at 0 h after CLP surgery, the levels of I-A/I-E and CD80 in mouse cells at 6, 12, 24, 48, and 72 h after CLP surgery and the levels of CD86 in mouse cells at 12, 24, and 48 h after CLP surgery were significantly increased ( P<0.05). At 24 h post-surgery, the protein expressions of TXNRD1, SLC7A11, and GPX4 in mouse cells in corn oil+CLP group were significantly lower than those in corn oil+sham injury group ( P<0.05), while the protein expressions of TXNRD1, SLC7A11, and GPX4 in mouse cells in inhibitor+CLP group were significantly lower than those in corn oil+CLP group and inhibitor+sham injury group ( P<0.05). At 24 h post-surgery, the content of GSH in mouse cells in corn oil+CLP group was (239±32) μg/mg, which was significantly lower than (366±59) μg/mg in corn oil +sham injury group ( P<0.05); the content of GSH in mouse cells in inhibitor+CLP group was (134±19) μg/mg, which was significantly lower than (355±31) μg/mg in inhibitor+sham injury group and that in corn oil+CLP group (with both P values <0.05). At 24 h post-surgery, the lipid peroxidation level of mouse cells in inhibitor+CLP group was significantly higher than that in the other three groups ( P<0.05). At 24 h post-surgery, the levels of I-A/I-E, CD80, and CD86 in mouse cells in corn oil+CLP group were significantly higher than those in corn oil+sham injury group ( P<0.05), while the levels of I-A/I-E and CD80 in mouse cells in inhibitor+CLP group were significantly higher than those in inhibitor+sham injury group ( P<0.05) but significantly lower than those in corn oil+CLP group ( P<0.05); the level of CD86 in mouse cells in inhibitor+sham injury group was significantly higher than that in corn oil+sham injury group ( P<0.05). Within 7 d post-surgery, the survival rate of mice in inhibitor+CLP group was significantly lower than that in inhibitor+sham injury group and corn oil+CLP group (with χ2 values of 31.19 and 3.91, respectively, both P values <0.05). Conclusions:In septic mice, the expression of TXNRD1 in DCs is reduced, cell ferroptosis is enhanced, and immune function is weakened. The inhibition of TXNRD1 in DCs will exacerbate cell ferroptosis and immune function suppression, and is closely related to the poor prognosis of sepsis.
2.Effect of the multidisciplinary management model on the prognosis of patients with portal hypertension
Changxiang LAI ; Qiyuan TANG ; Zhiyu LI ; Yujin JIANG ; Xuan ZHOU ; Wenjie XIAN ; Ruikun CHEN ; Xiaojuan WU ; Qing HE ; Fang WANG
Journal of Clinical Hepatology 2025;41(10):2068-2074
ObjectiveTo investigate the effect of the multidisciplinary team (MDT) management model in improving the prognosis of patients with cirrhotic portal hypertension. MethodsA total of 86 patients with cirrhotic portal hypertension who were admitted to Shenzhen Third People’s Hospital from May 2022 to July 2024 were enrolled, and according to whether the MDT treatment regimen was implemented, they were divided into execution group with 51 patients and non-execution group with 35 patients. Baseline clinical data were collected, and the patients were observed in terms of gastrointestinal bleeding, hepatic encephalopathy, liver cancer, and death from admission to the end of follow-up (January 2025). The independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups. The Kaplan-Meier method was used to plot survival curves for the cumulative incidence rates of endpoint events (gastrointestinal bleeding, hepatic encephalopathy, liver cancer, and death), and the Log-rank test was used for comparison between groups. The Cox proportional-hazards regression model analysis was used to investigate the effect of MDT management on the prognosis of patients. ResultsThere were significant differences between the execution group and the non-execution group in diameter of the portal vein (t=1.216, P=0.017) and ascites (χ2=4.515, P=0.034) at baseline. The patients were followed up for 14.6±6.2 months, and the survival curve analysis showed that there was a significant difference in the cumulative incidence rate of gastrointestinal bleeding between the two groups (χ2=4.573, P=0.024), while there were no significant differences in the incidence rates of other outcome events between the two groups (all P>0.05). The Cox regression analysis showed that the execution group had a reduced risk of gastrointestinal bleeding (hazard ratio=0.262, 95% confidence interval: 0.110 — 0.630, P=0.003). ConclusionImplementation of the MDT treatment regimen can significantly reduce the short-term risk of gastrointestinal bleeding in patients with cirrhotic portal hypertension, while its long-term benefits require further follow-up verification.
3.Analysis of the Main Chemical Components and Seasonal Differences of Mussels
Xiaozheng HUANG ; Qianqian ZHONG ; Xinyu PAN ; Wanglin BAO ; Qiyuan FENG ; Jizhou YIN ; Jiangyu LI-ANG ; Shenghan YAN ; Jidong ZHOU ; Jipeng SUN ; Hao WU ; Rui LIU
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(3):383-392
OBJECTIVE To analyze the differences in chemical composition of mussels from Shengsi,Zhoushan in different har-vest months,and to compare the differences in the composition of mussels from different origins,so as to provide ideas for the compre-hensive utilization of its resources.METHODS Ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UHPLC-QqQ-MS)and spectrophotometry were used to analyze and evaluate the contents of nucleosides,amino acids,water-soluble proteins,total sugars and polysaccharides in mussels from Shengsi,Zhoushan in 12 months of the year and from 5 different origins;principal component analysis(PCA),partial least squares discriminant method(PLS-DA)and TOPSIS method were used to compre-hensively evaluate mussels.RESULTS A total of 16 amino acids and 11 nucleosides were detected in mussels.The average content of total amino acids in Shengsi mussels throughout the year was 4 851.74 μg·g-1,the average content of total nucleosides was 921.40 μg·g-1,and the average contents of water-soluble protein,polysaccharides,and total sugars were 51.32 mg·g-1,74.39 μg·g-1,and 417.22 mg·g-1,respectively.The nucleosides of Shengsi mussels were the highest in March and April,the ami-no acids and water-soluble proteins were the highest in April and May,and the polysaccharides and total sugars were the highest in Oc-tober and November.The PCA and PLS-DA results of the chemical components of different resources showed that there were great differences in Shengsi mussel samples harvested in different seasons.The entropy weight TOPSIS analysis showed that the comprehen-sive scores of Shengsi mussels in March and April(S7 and S8)were better than those in other areas,and the comprehensive score of Shengsi mussels in April(S8)was the highest,which was determined to be the best harvesting month.CONCLUSION The comparison results of multi-type resource chemistry show that there are great differences in the chemical compo-sition of Shengsi mussels in different harvesting seasons;the quality of Shengsi mussels is better than that of other areas;April is the best harvesting season,providing a reference for the quality evaluation and comprehensive development and utilization of mussels.
4.The current research status and prospect of the pathogenesis,influencing factors and surgical prevention and treatment strategies of inguinal hernia after prostate cancer surgery
Qiyuan LUO ; Hongchao SHAO ; Shaohua ZHOU ; Mushi YE
Practical Oncology Journal 2025;39(5):445-449
Prostate cancer is one of the common malignant tumors in men.With the aging of the population and the progress of diagnostic technology,its incidence continues to rise.Radical prostatectomy is an effective treatment for localized prostate cancer.However,the incidence of inguinal hernia after surgery is high,which seriously affects patients' postoperative recovery and quality of life.This article provides a systematic review for the pathogenesis of inguinal hernia after prostate cancer surgery,including changes in anatomical structure,scar formation in the retroperitoneal space,no closure of sheath process,and increased intra-abdominal pressure after surgery.It also summarizes the relevant influencing factors,such as surgical methods,patients' own conditions,and postoperative recovery status.On this basis,surgical prevention and treatment strategies are explored,such as intraoperative preventive repair,re-fined surgical procedures,and postoperative targeted treatment.At the same time,future research directions are proposed,focusing on the in-depth exploration of molecular biological mechanisms,research and development of new materials,and application of artificial intelligence technology,aiming to reduce the incidence of inguinal hernia after prostate cancer surgery and improving patients' postop-erative quality of life.
5.The current research status and prospect of the pathogenesis,influencing factors and surgical prevention and treatment strategies of inguinal hernia after prostate cancer surgery
Qiyuan LUO ; Hongchao SHAO ; Shaohua ZHOU ; Mushi YE
Practical Oncology Journal 2025;39(5):445-449
Prostate cancer is one of the common malignant tumors in men.With the aging of the population and the progress of diagnostic technology,its incidence continues to rise.Radical prostatectomy is an effective treatment for localized prostate cancer.However,the incidence of inguinal hernia after surgery is high,which seriously affects patients' postoperative recovery and quality of life.This article provides a systematic review for the pathogenesis of inguinal hernia after prostate cancer surgery,including changes in anatomical structure,scar formation in the retroperitoneal space,no closure of sheath process,and increased intra-abdominal pressure after surgery.It also summarizes the relevant influencing factors,such as surgical methods,patients' own conditions,and postoperative recovery status.On this basis,surgical prevention and treatment strategies are explored,such as intraoperative preventive repair,re-fined surgical procedures,and postoperative targeted treatment.At the same time,future research directions are proposed,focusing on the in-depth exploration of molecular biological mechanisms,research and development of new materials,and application of artificial intelligence technology,aiming to reduce the incidence of inguinal hernia after prostate cancer surgery and improving patients' postop-erative quality of life.
6.Analysis of the Main Chemical Components and Seasonal Differences of Mussels
Xiaozheng HUANG ; Qianqian ZHONG ; Xinyu PAN ; Wanglin BAO ; Qiyuan FENG ; Jizhou YIN ; Jiangyu LI-ANG ; Shenghan YAN ; Jidong ZHOU ; Jipeng SUN ; Hao WU ; Rui LIU
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(3):383-392
OBJECTIVE To analyze the differences in chemical composition of mussels from Shengsi,Zhoushan in different har-vest months,and to compare the differences in the composition of mussels from different origins,so as to provide ideas for the compre-hensive utilization of its resources.METHODS Ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UHPLC-QqQ-MS)and spectrophotometry were used to analyze and evaluate the contents of nucleosides,amino acids,water-soluble proteins,total sugars and polysaccharides in mussels from Shengsi,Zhoushan in 12 months of the year and from 5 different origins;principal component analysis(PCA),partial least squares discriminant method(PLS-DA)and TOPSIS method were used to compre-hensively evaluate mussels.RESULTS A total of 16 amino acids and 11 nucleosides were detected in mussels.The average content of total amino acids in Shengsi mussels throughout the year was 4 851.74 μg·g-1,the average content of total nucleosides was 921.40 μg·g-1,and the average contents of water-soluble protein,polysaccharides,and total sugars were 51.32 mg·g-1,74.39 μg·g-1,and 417.22 mg·g-1,respectively.The nucleosides of Shengsi mussels were the highest in March and April,the ami-no acids and water-soluble proteins were the highest in April and May,and the polysaccharides and total sugars were the highest in Oc-tober and November.The PCA and PLS-DA results of the chemical components of different resources showed that there were great differences in Shengsi mussel samples harvested in different seasons.The entropy weight TOPSIS analysis showed that the comprehen-sive scores of Shengsi mussels in March and April(S7 and S8)were better than those in other areas,and the comprehensive score of Shengsi mussels in April(S8)was the highest,which was determined to be the best harvesting month.CONCLUSION The comparison results of multi-type resource chemistry show that there are great differences in the chemical compo-sition of Shengsi mussels in different harvesting seasons;the quality of Shengsi mussels is better than that of other areas;April is the best harvesting season,providing a reference for the quality evaluation and comprehensive development and utilization of mussels.
7.Effects of thioredoxin reductase 1 on ferroptosis and immune function of dendritic cells in septic mice
Qiyuan ZHOU ; Jingyan LI ; Yanmin CAO ; Weiling LI ; Ning DONG ; Yao WU ; Yingping TIAN ; Yongming YAO
Chinese Journal of Burns 2025;41(3):212-221
Objective:To investigate the effects of thioredoxin reductase 1 (TXNRD1) on ferroptosis and immune function of dendritic cells (DCs) in septic mice, and to provide a basis for improving the immunosuppression in sepsis caused by wound infection.Methods:This study was an experimental research. Sixty male C57BL/6J mice aged 6-8 weeks were subjected to cecal ligation and puncture (CLP) to establish sepsis models. Ten mice were selected at 0 (immediately), 6, 12, 24, 48, and 72 h after CLP surgery, respectively, according to the random number table method. Mouse splenic DCs were isolated using CD11c-positive magnetic beads. The protein expressions of TXNRD1, and anti-ferroptosis proteins solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the cells were detected by Western blotting, the reduced glutathione (GSH) content in the cells was measured by colorimetric assay, the lipid peroxidation level was assessed via live-cell imaging technology, and the levels of major histocompatibility complex class Ⅱ subtype I-A/I-E and leukocyte differentiation antigens CD80 and CD86 were detected by flow cytometry. Another 100 male C57BL/6J mice aged 6-8 weeks were divided into corn oil+sham injury group, corn oil+CLP group, inhibitor+sham injury group, and inhibitor+CLP group according to the random number table method, with 25 mice in each group. Mice in the two inhibitor groups were intraperitoneally injected with TXNRD1 inhibitor auranofin, while mice in the two corn oil groups were intraperitoneally injected with corn oil. One hour later, mice in the two CLP groups underwent CLP surgery to establish sepsis models, while mice in the two sham injury groups underwent sham surgery. Twenty mice from each group were selected to observe survival within 7 d post-surgery, and the survival rate was calculated. At 24 h post-surgery, mouse splenic DCs from the remaining 5 mice in each group were collected for corresponding assays as above.Results:Compared with those at 0 h after CLP surgery, the protein expressions of TXNRD1, GPX4, and SLC7A11 in mouse cells at 24 h after CLP surgery and the protein expression of TXNRD1 in mouse cells at 48 h after CLP surgery were significantly decreased ( P<0.05), the GSH content in mouse cells was significantly decreased at 24 and 48 h after CLP surgery ( P<0.05). The lipid peroxidation level in mouse cells was low at 0, 6, and 12 h after CLP surgery, slightly lower than that at 72 h after CLP surgery; the lipid peroxidation levels in mouse cells at 24 and 48 h after CLP surgery were significantly higher than those at 0, 6, 12, and 72 h after CLP surgery. Compared with those at 0 h after CLP surgery, the levels of I-A/I-E and CD80 in mouse cells at 6, 12, 24, 48, and 72 h after CLP surgery and the levels of CD86 in mouse cells at 12, 24, and 48 h after CLP surgery were significantly increased ( P<0.05). At 24 h post-surgery, the protein expressions of TXNRD1, SLC7A11, and GPX4 in mouse cells in corn oil+CLP group were significantly lower than those in corn oil+sham injury group ( P<0.05), while the protein expressions of TXNRD1, SLC7A11, and GPX4 in mouse cells in inhibitor+CLP group were significantly lower than those in corn oil+CLP group and inhibitor+sham injury group ( P<0.05). At 24 h post-surgery, the content of GSH in mouse cells in corn oil+CLP group was (239±32) μg/mg, which was significantly lower than (366±59) μg/mg in corn oil +sham injury group ( P<0.05); the content of GSH in mouse cells in inhibitor+CLP group was (134±19) μg/mg, which was significantly lower than (355±31) μg/mg in inhibitor+sham injury group and that in corn oil+CLP group (with both P values <0.05). At 24 h post-surgery, the lipid peroxidation level of mouse cells in inhibitor+CLP group was significantly higher than that in the other three groups ( P<0.05). At 24 h post-surgery, the levels of I-A/I-E, CD80, and CD86 in mouse cells in corn oil+CLP group were significantly higher than those in corn oil+sham injury group ( P<0.05), while the levels of I-A/I-E and CD80 in mouse cells in inhibitor+CLP group were significantly higher than those in inhibitor+sham injury group ( P<0.05) but significantly lower than those in corn oil+CLP group ( P<0.05); the level of CD86 in mouse cells in inhibitor+sham injury group was significantly higher than that in corn oil+sham injury group ( P<0.05). Within 7 d post-surgery, the survival rate of mice in inhibitor+CLP group was significantly lower than that in inhibitor+sham injury group and corn oil+CLP group (with χ2 values of 31.19 and 3.91, respectively, both P values <0.05). Conclusions:In septic mice, the expression of TXNRD1 in DCs is reduced, cell ferroptosis is enhanced, and immune function is weakened. The inhibition of TXNRD1 in DCs will exacerbate cell ferroptosis and immune function suppression, and is closely related to the poor prognosis of sepsis.
8.Ultra-fast-track anesthesia management for surgeries for acute type A aortic dissection
Yifan ZOU ; Qiyuan LIU ; Xu SUN ; Yi ZHOU ; Yun REN ; Hao YAO
Chinese Journal of Anesthesiology 2024;44(5):574-578
The clinical data from 16 patients who underwent surgery for acute type A aortic dissection under ultra-fast-track anesthesia (UFTA) in the Second Affiliated Hospital of Nanjing Medical University from August 2023 to December 2023 were collected, and the experience of anesthesia management was analyzed and summarized. UFTA strategy was to reduce inflammation and perform multi-organ protection in the perioperative period through cardiopulmonary bypass methods, medications, etc. According to the process of operation, the long-term and short-acting anesthetic drugs were accurately used to achieve the patient′s awakening immediately after the operation. Deep anesthesia extubation, remifentanil infusion techniques, and nasal high-flow ventilation were used to enable patients to be removed from the endotracheal tube with low stress under the premise of safety. All the patients successfully completed anesthesia induction and anesthesia maintenance. The average surgery duration was (413±92) min, the average anesthesia duration was (480±100) min, the cardiopulmonary bypass time was (168±42) min, the cardiac arrest time was (119±36) min, and the deep hypothermic circulatory arrest lasted (24±5) min. Of these patients, 14 underwent deep hypothermic circulatory arrest combined with unilateral cerebral perfusion via the right axillary artery, and 2 underwent deep hypothermic circulatory arrest combined with retrograde cerebral perfusion via the superior vena cava. All the patients had their tracheal tubes removed within 30 min postoperatively, with blood gas analysis and hemodynamics being within or close to normal ranges. The average postoperative intensive care unit stay time was 6 (3, 11) days, and the total length of hospital stay was (23±8) days. The total hospitalization time extended 20 days in 7 patients due to postoperative complications. All the patients recovered and were discharged. In conclusion, the application of UFTA strategy is safe and feasible in the patients with acute type A aortic dissection, allowing for immediate extubation in the operating room.
9.Role of apurinic/apyrimidinic endodeoxyribonuclease 1 in ferroptosis of mouse dendritic cells under simulated sepsis
Qiyuan ZHOU ; Jingyan LI ; Yongming YAO ; Yingping TIAN
Chinese Journal of Burns 2024;40(10):930-939
Objective:To investigate the role of apurinic/apyrimidinic endodeoxyribonuclease 1 (APE1) in ferroptosis of mouse dendritic cells (DCs) under simulated sepsis, providing evidence for improving immunosuppression in sepsis caused by wound infection.Methods:This study was an experimental research. The mouse DC line DC2.4 in the logarithmic growth phase (with passages 3-10) was used for the experiments (with each sample size of 3). The sepsis model was established by treating DCs with 1 μg/mL lipopolysaccharide (LPS, same concentration throughout) for 0 (untreated), 6, 12, 24, 48, and 72 h. Western blotting was used to detect the protein expressions of APE1 and anti-ferroptosis proteins glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) in cells, flow cytometry was employed to detect reactive oxygen species (ROS) levels in cells, and live-cell imaging technology was used to measure cell lipid peroxidation levels. DCs successfully transfected with lentivirus containing APE1 short hairpin RNA sequence were divided into APE1-knockdown+phosphate buffer solution (PBS) group and APE1-knockdown+LPS group. DCs successfully transfected with empty lentivirus were divided into empty vector+PBS group and empty vector+LPS group. After stimulation with PBS or LPS and 24 h of culture, corresponding assays were conducted as above. DCs transfected with lentivirus containing APE1 overexpression RNA sequence were divided into APE1-overexpression+PBS group and APE1-overexpression+LPS group. DCs transfected with empty lentivirus were divided into empty vector+PBS group and empty vector+LPS group. After stimulation with PBS or LPS and 24 h of culture, corresponding assays were conducted as above. A total of 88 male C57BL/6J mice aged 6-8 weeks were divided into corn oil+sham injury group, corn oil+cecal ligation and puncture (CLP) group, inhibitor+sham injury group, and inhibitor+CLP group ( n=22) according to the random number table. Mice in the two inhibitor groups were gavaged with APE1 inhibitor E3330 (1 mg/mL in concentration) at 40 mg/kg per day, while mice in the two corn oil groups were gavaged with an equal amount of corn oil daily. Two weeks later, mice in the two CLP groups underwent CLP surgery to establish a sepsis model, while mice in the two sham injury groups underwent sham injury. Sixteen mice from each group were selected to observe survival within 7 d post-surgery. At 24 h post-surgery, CD11c-positive magnetic beads were used to extract spleen DCs from the remaining six mice in each group for corresponding assays ( n=3) as above. Results:Compared with that of LPS treatment at 0 h, APE1 protein expression significantly increased in cells at 6 h of LPS treatment ( P<0.05), APE1 and GPX4 protein expressions significantly decreased at 24, 48, and 72 h of LPS treatment, SLC7A11 protein expression significantly decreased at 24 h of LPS treatment ( P<0.05), while the ROS level in cells ( P<0.05) and cell lipid peroxidation level significantly increased at 24, 48, and 72 h of LPS treatment. After 24 h of culture, the GPX4 protein expression of cells in APE1-knockdown+LPS group was significantly lower than that in APE1-knockdown+PBS group ( P<0.05), while the ROS level in cells ( P<0.05) and cell lipid peroxidation level were significantly higher than those in APE1-knockdown+PBS group and empty vector+LPS group. After 24 h of culture, APE1, SLC7A11, and GPX4 protein expressions of cells in APE1-overexpression+LPS group were significantly higher than those in empty vector+LPS group ( P<0.05), while the ROS level in cells ( P<0.05) and cell lipid peroxidation level were significantly lower than those in empty vector+LPS group. At 24 h post-surgery, APE1 and GPX4 protein expressions of mice cells in inhibitor+CLP group were significantly lower than those in inhibitor+sham injury group and corn oil+CLP group ( P<0.05); the ROS level of mice cells in corn oil+CLP group (12 693±913) was significantly higher than that in corn oil+sham injury group (4 205±805, P<0.05), while the ROS level of mice cells in inhibitor+CLP group (18 085±223) was significantly higher than that in inhibitor+sham injury group (4 381±787) and corn oil+CLP group (with P values all <0.05); the cell lipid peroxidation level of mice in inhibitor+CLP group was significantly higher than that in inhibitor+sham injury group and corn oil+CLP group. Within 7 d post-surgery, the survival ratio of mice in inhibitor+CLP group was significantly lower than that in inhibitor+sham injury group ( χ2=22.67, P<0.05). Conclusions:Under simulated sepsis, the APE1 expression in mouse DCs is decreased, and oxidative stress and ferroptosis are enhanced; knocking down the APE1 exacerbates DC ferroptosis, while APE1 overexpression effectively reduces DC ferroptosis. The inhibition of APE1 expression in DCs is closely associated with poor prognosis in sepsis.
10.Effects of stimulator of interferon gene on ferroptosis mediated by acyl-CoA synthetase long-chain family member 4 in mouse dendritic cells under sepsis
Mengyao WU ; Pengyi HE ; Yu DUAN ; Liyu ZHENG ; Renqi YAO ; Qiyuan ZHOU ; Yu CHEN ; Ning DONG ; Yao WU ; Yongming YAO
Chinese Journal of Burns 2024;40(10):920-929
Objective:To investigate the effects of stimulator of interferon gene (STING) on ferroptosis mediated by acyl-CoA synthetase long-chain family member 4 (ACSL4) in mouse dendritic cells (DCs) under sepsis, providing a basis for improving the dysregulation of immune response in sepsis caused by factors such as wound infection.Methods:This study was an experimental research. The mouse DC line DC2.4 in the logarithmic growth phase (with passages 3-10) were divided into lipopolysaccharide (LPS) stimulation 0 h (unstimulated) group, LPS stimulation 6 h group, LPS stimulation 12 h group, LPS stimulation 18 h group, and LPS stimulation 24 h group according to the random number table (the same grouping method below), which were cultured with 1 μg/mL LPS (the same concentration below) for the corresponding time. The protein expressions of phosphorylated STING (p-STING), STING, and ACSL4 in cells were determined by Western blotting. DC2.4 successfully transfected with lentivirus containing STING gene small interfering RNA (hereinafter referred to as siSTING) were divided into siSTING+phosphate buffer solution (PBS) group and siSTING+LPS group. DC2.4 successfully transfected with empty lentivirus were divided into empty vector+PBS group and empty vector+LPS group. After being stimulated with PBS or LPS and cultured for 24 hours, the protein expressions of p-STING, STING, and ACSL4 in cells were determined as above. Cell lipid peroxidation degrees were observed using the lipid peroxidation assay kit, and cell apoptosis rates were detected using flow cytometry. The sample numbers in the above cell experiments were all 3. Eighty male C57BL/6J mice aged 6 to 8 weeks were divided into sham surgery+normal saline (NS) group, cecal ligation and puncture (CLP)+NS group, sham surgery+C-176 group, and CLP+C-176 group, with 20 mice in each group. Mice in the two C-176 groups were intraperitoneally injected with C-176, while mice in the two NS groups were intraperitoneally injected with an equivalent volume of NS. One hour later, sham surgery was performed on the mice in the two sham surgery groups, and CLP surgery was performed on the mice in the two CLP groups to establish a sepsis model. At 24 h post-surgery, 10 mice from each group were sacrificed to extract spleen DCs, and protein expression, lipid peroxidation, and apoptosis rates were detected as above ( n=3). Hematoxylin-eosin staining was performed to observe pathological damage in the heart, liver, lung, and kidney tissue. The remaining 10 mice in each group were observed for survival within 7 days after surgery. Results:The protein expressions of p-STING, STING, and ACSL4, as well as the p-STING/STING ratio in DC2.4 in LPS stimulation 24 h group were significantly higher than those in LPS stimulation 0 h group ( P<0.05). After 24 h of culture, the protein expressions of p-STING, STING, and ACSL4 in DC2.4 in siSTING+LPS group and empty vector+PBS group were significantly lower than those in empty vector+LPS group ( P<0.05); the lipid peroxidation degrees of DC2.4 in siSTING+LPS group and empty vector+PBS group were weaker than those in empty vector+LPS group. The apoptosis rates of DC2.4 in empty vector+PBS group, empty vector+LPS group, siSTING+PBS group, and siSTING+LPS group were (15.7±3.0)%, (37.8±2.9)%, (13.1±2.1)%, and (20.6±1.8)%, respectively. The apoptosis rates of DC2.4 in empty vector+PBS group and siSTING+LPS group were significantly lower than that in empty vector+LPS group ( P<0.05). At 24 h post-surgery, the protein expressions of p-STING and ACSL4, and the p-STING/STING ratio in spleen DCs of mice in CLP+NS group were significantly higher than those in sham surgery+NS group and CLP+C-176 group ( P<0.05); the protein expression of STING in spleen DCs of mice in CLP+NS group was significantly higher than that in sham surgery+NS group ( P<0.05); the lipid peroxidation degrees of spleen DCs of mice in CLP+C-176 group and sham surgery+NS group were weaker than that in CLP+NS group. The apoptosis rates of spleen DCs of mice in sham surgery+NS group and CLP+C-176 group were significantly lower than that in CLP+NS group ( P<0.05), and the apoptosis rate of spleen DCs of mice in CLP+C-176 group was significantly higher than that in sham surgery+C-176 group ( P<0.05). Pathological tissue damage in the heart, liver, lung, and kidney of mice in CLP+NS group was significantly worse than that in sham surgery+NS group, while such damage in the above organs of mice in CLP+C-176 group was significantly alleviated compared with that in CLP+NS group. The survival ratio of mice in CLP+NS group within 7 days after surgery was significantly lower than that in sham surgery+NS group ( χ2=8.30, P<0.05). Conclusions:Under sepsis, STING activation in mouse DCs is significant, which enhances ACSL4-mediated ferroptosis. Inhibiting STING activation can significantly reduce ACSL4-mediated ferroptosis level in mouse DCs under sepsis, thereby improving the survival rate of septic mice.

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