1.A Single-cell Transcriptomic Study of Wenyang Jiedu Granules in Alleviating Influenza Virus Pneumonia by Suppressing Interferon Responses and Apoptotic Pathways in AT1 Cells
Jingyan XIN ; Shengle QIN ; Taoyu CHEN ; Yuntao LIU ; Xiaolu WANG ; Chaofeng LIANG ; Zhongde ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):76-85
ObjectiveTo investigate the molecular mechanisms by which Wenyang Jiedu granules (WYJD) alleviate influenza A virus (IAV)-induced pneumonia based on single-cell transcriptome sequencing. MethodsThirty female BALB/c mice were randomly divided into a blank control group (Control), IAV group, and WYJD low-, medium-, and high-dose groups (WYJD-L, WYJD-M, WYJD-H; 2.925, 5.85, 11.7 g·kg-1, n=6). Except for the Control group, all other groups were intranasally inoculated with IAV subtype H1N1 (A/PR/8/34) to establish an infection model. Two hours after modeling, drug administration was initiated and continued for 5 consecutive days, with daily monitoring of body weight and general condition. On day 6, mice were sacrificed and samples were collected. Lung index was calculated, and histopathological examination of lung tissue was performed. Lung tissues from the Control, IAV, and WYJD-H groups were subjected to single-cell transcriptome sequencing (n=3), focusing on type I alveolar epithelial cells (AT1) to analyze changes in gene expression and signaling pathways. Western blot was used to detect the expression changes of relevant proteins to validate the single-cell sequencing results. ResultsCompared with the Control group, the IAV group exhibited significantly decreased body weight (P<0.05) and significantly increased lung index (P<0.05). Compared with the IAV group, all WYJD-treated groups exhibited significantly increased body weight (P<0.01) and significantly decreased lung index (P<0.01). Single-cell sequencing analysis revealed that WYJD inhibited overactivation of interferon and inflammatory signaling pathways in AT1 cells after IAV infection, including interferon-γ response, interferon-α response, tumor necrosis factor-α/nuclear factor-κB (TNF-α/NF-κB), and interleukin-6/Janus kinase/signal transducer and activator of transcription 3 (IL-6/JAK/STAT3) pathways. Compared with the Control group, the number of AT1 cells in the IAV group showed a decreasing trend. Compared with the IAV group, the WYJD-H group showed an increasing trend, although neither difference was statistically significant. Further analysis of AT1 cell subpopulation gene expression showed that, compared with the Control group, the IAV group exhibited increased expression of pro-apoptotic genes FAS cell surface death receptor (FAS) and cyclin-dependent kinase inhibitor 1A (CDKN1A), a significant increase in tumor protein p53 (Tp53) expression (P<0.05), and significant decreases in expression of the AT1 marker gene advanced glycosylation end-product-specific receptor (AGER) and membrane structural gene caveolin1 (CAV1) (P<0.05, P<0.01). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of FAS, CDKN1A, and Tp53 (P<0.05, P<0.01), and significantly increased expression of AGER and CAV1 (P<0.05, P<0.01). Regarding interferon response-related genes, compared with the Control group, the IAV group showed increased expression of interferon-stimulated gene 15 (ISG15), interferon-induced protein with tetratricopeptide repeats 3 (IFIT3), signal transducer and activator of transcription 2 (STAT2), bone marrow stromal cell antigen 2 (BST2), and C-X-C motif chemokine ligand 10 (CXCL10), with a significant increase in 2′,5′-oligoadenylate synthetase-like protein 1 (OASL1) (P<0.05). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of all the above genes, with highly significant differences for ISG15, IFIT3, STAT2, BST2, and OASL1 (P<0.01), and a significant difference for CXCL10 (P<0.05). Among inflammation-related genes, compared with the Control group, the IAV group showed significantly increased expression of intercellular adhesion molecule 1 (ICAM1), tumor necrosis factor alpha-induced protein 3 (TNFAIP3), keratin 8 (KRT8), tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), and TNFRSF1B (P<0.01), and increased expression of NFKBIA, a negative regulator of NF-κB (P<0.05). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of KRT8 and TNFRSF1B (P<0.05), while ICAM1, NFKBIA, TNFAIP3, and TNFRSF1A showed decreasing trends without statistical significance. Western blot validation showed that, compared with the Control group, protein expression levels of ISG15, FAS, p53, and phosphorylated p65 (p-p65) in lung tissue of the IAV group were significantly increased (P<0.05, P<0.01). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of these proteins (P<0.05, P<0.01). ConclusionWYJD may alleviate viral pneumonia by targeting gene expression in AT1 cells, inhibiting overactivated interferon and inflammatory signaling pathways after IAV infection, and downregulating pro-apoptotic signaling, thereby reducing alveolar epithelial injury.
2.Clinical efficacy analysis of autologous rib cartilage reconstruction for finger hemiarticular defects
Zhiyu HU ; Chaofeng XING ; Tao YANG ; Jia CHEN ; Zirun XIAO ; Xiazhi LIU ; Li SONG ; Beibei CHENG ; Yingjie XIONG ; Guangchao ZHANG ; Songfeng YANG
Journal of Clinical Surgery 2025;33(6):641-645
Objective Exploring the clinical efficacy of using autologous rib cartilage grafting to reconstruct finger hemiarticular defects.Methods From August 2022 to February 2024,for 6 patients with hemiarticular surface defects in the metacarpophalangeal joints and interphalangeal joints of 8 fingers,costal cartilage was used for joint remodeling and transplantation to reconstruct the hemiarticular surface defects of the fingers.All 8 joint transplants in 6 patients were two-stage surgeries.In the first stage,antibiotic bone cement was used to fill the space-occupying lesions,and in the second stage,costal cartilage joint remodeling was performed to reconstruct the finger joint defects.Postoperative follow-up and assessment of fracture healing according to Paley fracture healing scoring criteria.Outpatient and WeChat follow-up,upper limb function is evaluated according to the upper limb functional assessment standards of the Chinese Medical Association Hand Surgery Society.Record VAS pain score.Results In this group,there were a total of 6 patients with 8 cases of hemiarticular defects.Among them,2 patients had two joint surgical sites,while the remaining 4 patients had a single joint surgical site.There were 2 cases of metacarpophalangeal joint head defects,2 cases of proximal articular surface defects,3 cases of proximal articular head defects,and 1 case of thumb proximal articular head defect.After surgery,8 out of 6 patients'hand wounds healed successfully.All patients were followed up for 6-12 months postoperatively,with an average of 9 months.The VAS pain score(affected finger)for the last follow-up was 0-2 points,with an average of 0.6 points.According to Paley's scoring criteria,all 6 patients had excellent fracture healing.According to the evaluation criteria for upper limb functional assessment of the Chinese Medical Association Hand Surgery Society,3 cases were rated as excellent,3 cases were rated as good,and 2 cases were rated as fair.Conclusion For patients with half joint defects on one side of the finger,rib rib cartilage was used for joint reconstruction,which significantly improved the joint shape and function at the defect site,and reduced joint pain scores.
3.Intraosseous and periosteal arteries of hallux phalanx: a 3D observational study
Zirun XIAO ; Jia CHEN ; Kuo WEI ; Zhenfeng LI ; Chaofeng XING ; Weiguang ZHANG
Chinese Journal of Microsurgery 2025;48(5):547-551
Objective:To explore the feasibility of obtaining three-dimensional (3D) models of intraosseous and periosteal arteries of hallux phalanx using micro-arteriography with micro-CT scan.Methods:From January 2022 to April 2025, the Department of Orthopaedic, the 988th Hospital of the Joint Logistics Support Force of the Chinese PLA conducted a study on 7 fresh-frozen specimens of distal lower limb (right lower limb) from an 85-year-old male, and both lower limbs from an 82-year-old male, a 78-year-old female and a 66-year-old male in the Department of Human Anatomy & Histology and Embryology, Peking University School of Basic Medical Sciences. Red lead oxide powder (Pb 3O 4) was ground and filtered through a 300 mesh, and then mixed with turpentine at ratios of 1 g ∶ 1.5 ml, 1 g ∶ 1.0 ml, and 1 g ∶ 0.5 ml to prepare lead-based contrast agent suspensions. After thawing the specimens at room temperature, the suspensions were injected via the popliteal artery in ascending order of concentration. After injections, the specimens were fixed in 10% methanal for 2 weeks. The proximal and distal phalanges of the hallux, with the surrounding periosteum preserved intact, were then harvested. The harvested specimens were scanned using micro-CT at an ultimate resolution of 12 μm. Subsequently, Mimics Medical software was used to reconstruct 3D models of the intraosseous and periosteal arteries within the phalanges. Results:Periosteal arteries in the proximal phalanx were primarily distributed near the joint region. A consistently large trunk artery entered from plantar side, supplying most of the diaphysis and head. There was a rich periosteal arterial network on both sides of the distal phalanx, which communicates with each other through the arterial arch in the bone groove. However, trunk intraosseous artery could be absent. Intraosseous arteries in the proximal ends of both the proximal and distal phalanges originated from periosteal arteries. These formed an interconnected arteriosomes and coursed parallel to the articular surfaces.Conclusion:The micro-arteriography acquired by micro-CT scan effectively visualizes intraosseous and periosteal arteries and enables the reconstruction and analysis of 3D models of the arteriosomes. The characteristics of arteriolar distribution provide a theoretical basis for osteotomy or internal fixation procedures involving a hallux phalanx.
4.Recombinant human CC16 protein inhibits cigarette smoke extract-in-duced senescence-associated secretory phenotype in human bronchial epi-thelial cells and lung tissues from COPD mice
Kaiyan DU ; Ting LI ; Chaofeng LIU ; Xinyang LI ; Jingyu ZHANG ; Min GUO ; Zhaoyang CHEN ; Min PANG ; Hailong WANG
Chinese Journal of Pathophysiology 2025;41(2):294-302
AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P<0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P<0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P<0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P<0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P<0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P<0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P>0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.
5.Clinical efficacy analysis of autologous rib cartilage reconstruction for finger hemiarticular defects
Zhiyu HU ; Chaofeng XING ; Tao YANG ; Jia CHEN ; Zirun XIAO ; Xiazhi LIU ; Li SONG ; Beibei CHENG ; Yingjie XIONG ; Guangchao ZHANG ; Songfeng YANG
Journal of Clinical Surgery 2025;33(6):641-645
Objective Exploring the clinical efficacy of using autologous rib cartilage grafting to reconstruct finger hemiarticular defects.Methods From August 2022 to February 2024,for 6 patients with hemiarticular surface defects in the metacarpophalangeal joints and interphalangeal joints of 8 fingers,costal cartilage was used for joint remodeling and transplantation to reconstruct the hemiarticular surface defects of the fingers.All 8 joint transplants in 6 patients were two-stage surgeries.In the first stage,antibiotic bone cement was used to fill the space-occupying lesions,and in the second stage,costal cartilage joint remodeling was performed to reconstruct the finger joint defects.Postoperative follow-up and assessment of fracture healing according to Paley fracture healing scoring criteria.Outpatient and WeChat follow-up,upper limb function is evaluated according to the upper limb functional assessment standards of the Chinese Medical Association Hand Surgery Society.Record VAS pain score.Results In this group,there were a total of 6 patients with 8 cases of hemiarticular defects.Among them,2 patients had two joint surgical sites,while the remaining 4 patients had a single joint surgical site.There were 2 cases of metacarpophalangeal joint head defects,2 cases of proximal articular surface defects,3 cases of proximal articular head defects,and 1 case of thumb proximal articular head defect.After surgery,8 out of 6 patients'hand wounds healed successfully.All patients were followed up for 6-12 months postoperatively,with an average of 9 months.The VAS pain score(affected finger)for the last follow-up was 0-2 points,with an average of 0.6 points.According to Paley's scoring criteria,all 6 patients had excellent fracture healing.According to the evaluation criteria for upper limb functional assessment of the Chinese Medical Association Hand Surgery Society,3 cases were rated as excellent,3 cases were rated as good,and 2 cases were rated as fair.Conclusion For patients with half joint defects on one side of the finger,rib rib cartilage was used for joint reconstruction,which significantly improved the joint shape and function at the defect site,and reduced joint pain scores.
6.Analysis of clinical value of platelet antibody screening in 95 987 inpatients.
Ping CHEN ; Yang SUN ; Xiaoyue CHU ; Fenfang TIAN ; Yingqun YANG ; Wenhua WANG ; Jiameng NIU ; Boya ZHAO ; Jingyan CHANG ; Jiangcun YANG ; Chaofeng MA
Chinese Journal of Cellular and Molecular Immunology 2025;41(2):143-147
Objective To analyze the distribution of platelet antibodies in hospitalized patients and explore the clinical significance of platelet antibody detection. Methods A total of 95 987 hospitalized patient cases from a tertiary hospital in Xi'an from April 1, 2021 to December 31, 2023 were collected. Platelet antibodies were detected by solid-phase agglutination method. Statistical analysis was performed on variables including gender, age, blood type, department, history of blood transfusion, pregnancy history, and disease type. Results Among 95 987 hospitalized patients, the positive rate of platelet antibody detection reached 4.35%. The positive rate of platelet antibodies in female hospitalized patients (5.29%) was higher than that in male patients (3.31%), and the difference was statistically significant (x2=224.124). The positive rate of platelet antibodies in those with pregnancy history (7.92%) was higher than that in those without pregnancy history (4.19%), and the difference was significant (x2=292.773). Similarly, the positive rate of platelet antibodies in those with transfusion history (7.79%) was higher than that in those without transfusion history (3.97%), and the difference was significant (x2=300.209). There was a significant correlation between the positive rate of platelet antibodies and the number of pregnancies (x2=91.061). Conclusion The positive rate of platelet antibodies in 95 987 inpatient cases was 4.35%. The positive rate of platelet antibodies had a close relationship with a history of blood transfusions and pregnancies, and it increased with the number of pregnancies. For patients with multiple transfusion histories and pregnancy histories, screening for platelet antibodies holds significant diagnostic value.
Humans
;
Female
;
Male
;
Adult
;
Middle Aged
;
Blood Platelets/immunology*
;
Inpatients
;
Aged
;
Pregnancy
;
Young Adult
;
Adolescent
;
Aged, 80 and over
;
Autoantibodies/blood*
7.Recombinant human CC16 protein inhibits cigarette smoke extract-in-duced senescence-associated secretory phenotype in human bronchial epi-thelial cells and lung tissues from COPD mice
Kaiyan DU ; Ting LI ; Chaofeng LIU ; Xinyang LI ; Jingyu ZHANG ; Min GUO ; Zhaoyang CHEN ; Min PANG ; Hailong WANG
Chinese Journal of Pathophysiology 2025;41(2):294-302
AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P<0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P<0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P<0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P<0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P<0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P<0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P>0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.
8.Intraosseous and periosteal arteries of hallux phalanx: a 3D observational study
Zirun XIAO ; Jia CHEN ; Kuo WEI ; Zhenfeng LI ; Chaofeng XING ; Weiguang ZHANG
Chinese Journal of Microsurgery 2025;48(5):547-551
Objective:To explore the feasibility of obtaining three-dimensional (3D) models of intraosseous and periosteal arteries of hallux phalanx using micro-arteriography with micro-CT scan.Methods:From January 2022 to April 2025, the Department of Orthopaedic, the 988th Hospital of the Joint Logistics Support Force of the Chinese PLA conducted a study on 7 fresh-frozen specimens of distal lower limb (right lower limb) from an 85-year-old male, and both lower limbs from an 82-year-old male, a 78-year-old female and a 66-year-old male in the Department of Human Anatomy & Histology and Embryology, Peking University School of Basic Medical Sciences. Red lead oxide powder (Pb 3O 4) was ground and filtered through a 300 mesh, and then mixed with turpentine at ratios of 1 g ∶ 1.5 ml, 1 g ∶ 1.0 ml, and 1 g ∶ 0.5 ml to prepare lead-based contrast agent suspensions. After thawing the specimens at room temperature, the suspensions were injected via the popliteal artery in ascending order of concentration. After injections, the specimens were fixed in 10% methanal for 2 weeks. The proximal and distal phalanges of the hallux, with the surrounding periosteum preserved intact, were then harvested. The harvested specimens were scanned using micro-CT at an ultimate resolution of 12 μm. Subsequently, Mimics Medical software was used to reconstruct 3D models of the intraosseous and periosteal arteries within the phalanges. Results:Periosteal arteries in the proximal phalanx were primarily distributed near the joint region. A consistently large trunk artery entered from plantar side, supplying most of the diaphysis and head. There was a rich periosteal arterial network on both sides of the distal phalanx, which communicates with each other through the arterial arch in the bone groove. However, trunk intraosseous artery could be absent. Intraosseous arteries in the proximal ends of both the proximal and distal phalanges originated from periosteal arteries. These formed an interconnected arteriosomes and coursed parallel to the articular surfaces.Conclusion:The micro-arteriography acquired by micro-CT scan effectively visualizes intraosseous and periosteal arteries and enables the reconstruction and analysis of 3D models of the arteriosomes. The characteristics of arteriolar distribution provide a theoretical basis for osteotomy or internal fixation procedures involving a hallux phalanx.
9.Meta-synthesis of qualitative research on the psychological experience of non-suicidal self-injury in adolescents
Yuwei WU ; Yingying FAN ; Chaofeng LI ; Yi HUANG ; Xianglin KONG ; Dan CHEN
Chinese Journal of Modern Nursing 2024;30(26):3557-3565
Objective:To systematically integrate qualitative research on the psychological experience of non-suicidal self-injury (NSSI) in adolescents.Methods:A comprehensive literature search was conducted in databases including China National Knowledge Infrastructure, WanFang data, VIP, China Biology Medicine disc, PubMed, Web of Science, Embase, and Cochrane Library for qualitative studies on the psychological experience of NSSI in adolescents and the search period covered the inception of the databases until June 27, 2023. The methodological quality of the included studies was assessed using the Joanna Briggs Institute Critical Appraisal Checklist for Qualitative Research (2016 edition). The results were integrated using the Meta-synthesis method.Results:A total of 15 studies were included, comprising ten phenomenological studies, one descriptive qualitative study, one mixed-method study, one narrative study, one exploratory study, and one grounded theory study. Thirty-one findings were extracted and synthesized into ten categories, resulting in three main themes: triggers of adolescent NSSI, NSSI as a coping strategy, and the internal conflict and struggle of adolescents engaging in NSSI.Conclusions:Adolescents engaging in NSSI experience complex psychological and sensory responses. Healthcare professionals should closely monitor adolescents' emotional changes, enhance psychological assessments, and guide them in coping with negative emotions effectively to improve their physical and mental health.
10.Esophageal squamous cell carcinoma exosomal miR-181b-5p promotes tumor associated macrophage polarization by inhibiting PTEN
Hui FAN ; Naikuo ZHAO ; Linlin CHEN ; Zhiguo LI ; Miao LIU ; Yuanying ZHANG ; Chaofeng ZHOU
Chinese Journal of Immunology 2024;40(7):1441-1446
Objective:To investigate the effect and mechanism of esophageal squamous cell carcinoma exosomal miR-181b-5p to the polarization of M2 macrophages.Methods:Extracted and identified exosomes from esophageal squamous cell carcinoma,and detected the expression of miR-181b-5p in esophageal squamous cell carcinoma cells and their exosomes by qRT-PCR.M0 type macro-phages were divided into PBS group,HEEC exo group,Eca-109 exo group,miR-NC exo group,miR-181b-5p exo group,miR-NC group,miR-181b-5p mimic group,si-NC group,si-PTEN group,miR-181b-5p exo+PTEN group.qRT-PCR was used to detect the expressions of CD163,CD206,iNOS and TNF-α in each group.The targeting relationship between miR-181b-5p and PTEN were veri-fied by double luciferase reporter gene experiment.Results:miR-181b-5p was significantly overexpressed in esophageal squamous cell carcinoma cells TE-13,TE-12,TE-10,Eca-109,KYSE30 and their exosomes(P<0.001).Compared with miR-NC group,the expression of CD163 and CD206 in cells were significantly upregulated in the miR-181b-5p mimic group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).The results of double luciferase reporter genes showed that PTEN was the target gene of miR-181b-5p.Compared with si-NC group,the expressions of CD163 and CD206 in cells were significantly upregu-lated in the si-PTEN group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).Compared with PBS group,the expressions of CD163 and CD206 in cells were significantly upregulated in the Eca-109 exo group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).Compared with miR-NC exo group,the expressions of CD163 and CD206 in cells were significantly upregulated in the miR-181b-5p exo group,as well as the expressions of iNOS and TNF-α were significantly downregulated(P<0.001).Compared with miR-181b-5p exo group,the expressions of CD163 and CD206 in cells were significantly downregulated in the miR-181b-5p exo+PTEN group,as well as the expressions of iNOS and TNF-α were significantly upregulated(P<0.001).Conclusion:Exosomal miR-181b-5p inhibits PTEN expressions to promote M2 macrophage polarization.

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