1.Construction and efficacy verification of an intelligent pharmaceutical Q&A platform based on AI hallucination-suppression
Zhengwang WEN ; Jiaying WANG ; Wenyue YANG ; Haoyu YANG ; Xiao MA ; Yun LIU
China Pharmacy 2026;37(2):226-231
OBJECTIVE To construct an intelligent pharmaceutical Q&A platform for precision medication with low “artificial intelligence (AI) hallucination”, aiming to enhance the accuracy, consistency, and traceability of medication consultations. METHODS Medication package inserts were batch-processed and converted into structured data through Python programming to build a local pharmaceutical knowledge base. The retrieval and question-answering processes were designed based on large language models, and system integration and localized deployment were completed on Dify platform. By designing typical clinical medication questions and comparing the output of the intelligent pharmaceutical Q&A platform with the online version of DeepSeek across dimensions such as peak time retrieval, half-life, and dosage adjustment reasoning for patients with renal impairment, the accuracy and reliability of its retrieval and reasoning results were evaluated. RESULTS The intelligent pharmaceutical Q&A platform, constructed based on local drug package inserts, achieved 100% accuracy in retrieval and reasoning for peak time, half-life, and dosage adjustment schemes. In comparison, the online version of DeepSeek demonstrated accuracies of 30%(6/20), 50%(10/20), and 38%(23/60) across these three dimensions, respectively. CONCLUSIONS The constructed intelligent pharmaceutical Q&A platform is capable of accurately retrieving and extracting information from the local knowledge base based on clinical inquiries, thereby avoiding the occurrence of AI hallucinations and providing reliable medication decision support for healthcare professionals.
2.Clinical Observation on Tongyuan Acupuncture in the Treatment of Acute Gouty Arthritis
Lixian LI ; Minjie PAN ; Wenna ZHANG ; Qingxia XIE ; Zhongqi SHEN ; Jiaying XIAO ; Li CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(3):676-682
Objective To observe the clinical efficacy of Tongyuan acupuncture in the treatment of acute gouty arthritis.Methods A total of 126 cases of patients with definitive diagnosis of acute gouty arthritis admitted to the wards and outpatient clinics of Guangzhou Nansha District Hospital of Traditional Chinese Medicine from November 2023 to June 2024 were selected as the study subjects.The patients were randomly divided into observation group and control group according to the random number table method,with 63 cases in each group.The control group was given health training and conventional treatment,while the observation group was given Tongyuan acupuncture on the basis of treatment in the control group.Patients in both groups were treated for 14 consecutive days.After two weeks of treatment,the clinical efficacy of the two groups was evaluated.The changes in the clinical symptom scores,the Visual Analogue Scale(VAS)scores of pain,the levels of musculoskeletal ultrasonography(synovial proliferation,blood flow signals,and joint fluid)before and after the treatment were observed in patients of both groups.The changes of blood uric acid(UA),erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),tumor necrosis factor α(TNF-α),and interleukin 1(IL-1)before and after treatment were compared between the two groups.Results(1)The total effective rate was 95.24%(60/63)in the observation group and 84.13%(53/63)in the control group.The efficacy of the observation group was superior to that of the control group,the difference being statistically significant(P<0.05).(2)After treatment,the clinical symptom scores and VAS scores of the patients in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(3)After treatment,the serum UA,ESR,and CRP levels of patients in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(4)After treatment,the TNF-α and IL-1 levels of patients in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(5)After treatment,the musculoskeletal ultrasound indicators of the two groups of patients were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).Conclusion Tongyuan acupuncture in the treatment of acute gouty arthritis can significantly improve the clinical symptoms of patients,reduce the level of patients'blood UA and inflammatory factor.
3.Construction of predictive model for programmed death-1 inhibitor-related endocrine adverse events
Jiaying SHI ; Wei WEI ; Ting HAN ; Xiao ZHOU ; Meng ZHUO ; Xiaolin LIN ; Tao TAO ; Xiuying XIAO
Chinese Journal of Clinical Medicine 2025;32(4):551-560
Objective To identify the independent predictors of programmed death-1 (PD-1) inhibitor-related endocrine adverse events and construct a clinically usable risk prediction model. Methods A total of 302 patients with solid tumors treated with PD-1 inhibitors were retrospectively enrolled. According to the presence or absence of endocrine immune-related adverse events (irAEs), the patients were divided into case group and control group. The clinical and laboratory indexes were compared between the two groups. Multivariable logistic regression was used to confirm independent predictors of endocrine irAEs. The nomogram was constructed, while the receiver operating characteristic (ROC) curve was used to test the prediction performance of the model. Results The overall incidence of endocrine irAEs was 21.9% (66/302), and the incidence of hypothyroidism was 19.5% (59/302). The age, PD-1 inhibitors, free thyroxine, thyroid peroxidase antibody (TPOAb), thyroglobulin, amylase, lymphocyte subset CD3 expression were statistically different between the two groups (P<0.05). Multivariable logistic regression showed that higher expression of lymphocyte subset CD3 was a protective factor to prevent endocrine irAEs occurrence (P=0.004), while age<60 years, higher TPOAb and use of pembrolizumab were independent risk factors of endocrine irAEs (P<0.05). The nomogram model thus constructed, and when the threshold probability of the model exceeded 0.1, its net benefit was higher. ROC curve showed that the AUC of the model to predict endocrine irAEs was 0.760. The prediction result of the model was highly consistent with the actual result. Conclusions The age, type of PD-1 inhibitor, baseline TPOAb level, and baseline CD3 expression can independently predict endocrine irAEs occurrence or not. The nomogram model based on this model has good predictive efficiency, which can provide reference for early identification of high-risk patients and immunotherapy management.
4.POU2F1 inhibits miR-29b1/a cluster-mediated suppression of PIK3R1 and PIK3R3 expression to regulate gastric cancer cell invasion and migration.
Yizhi XIAO ; Ping YANG ; Wushuang XIAO ; Zhen YU ; Jiaying LI ; Xiaofeng LI ; Jianjiao LIN ; Jieming ZHANG ; Miaomiao PEI ; Linjie HONG ; Juanying YANG ; Zhizhao LIN ; Ping JIANG ; Li XIANG ; Guoxin LI ; Xinbo AI ; Weiyu DAI ; Weimei TANG ; Jide WANG
Chinese Medical Journal 2025;138(7):838-850
BACKGROUND:
The transcription factor POU2F1 regulates the expression levels of microRNAs in neoplasia. However, the miR-29b1/a cluster modulated by POU2F1 in gastric cancer (GC) remains unknown.
METHODS:
Gene expression in GC cells was evaluated using reverse-transcription polymerase chain reaction (PCR), western blotting, immunohistochemistry, and RNA in situ hybridization. Co-immunoprecipitation was performed to evaluate protein interactions. Transwell migration and invasion assays were performed to investigate the biological behavior of GC cells. MiR-29b1/a cluster promoter analysis and luciferase activity assay for the 3'-UTR study were performed in GC cells. In vivo tumor metastasis was evaluated in nude mice.
RESULTS:
POU2F1 is overexpressed in GC cell lines and binds to the miR-29b1/a cluster promoter. POU2F1 is upregulated, whereas mature miR-29b-3p and miR-29a-3p are downregulated in GC tissues. POU2F1 promotes GC metastasis by inhibiting miR-29b-3p or miR-29a-3p expression in vitro and in vivo . Furthermore, PIK3R1 and/or PIK3R3 are direct targets of miR-29b-3p and/or miR-29a-3p , and the ectopic expression of PIK3R1 or PIK3R3 reverses the suppressive effect of mature miR-29b-3p and/or miR-29a-3p on GC cell metastasis and invasion. Additionally, the interaction of PIK3R1 with PIK3R3 promotes migration and invasion, and miR-29b-3p , miR-29a-3p , PIK3R1 , and PIK3R3 regulate migration and invasion via the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway in GC cells. In addition, POU2F1 , PIK3R1 , and PIK3R3 expression levels negatively correlated with miR-29b-3p and miR-29a-3p expression levels in GC tissue samples.
CONCLUSIONS
The POU2F1 - miR-29b-3p / miR-29a-3p-PIK3R1 / PIK3R1 signaling axis regulates tumor progression and may be a promising therapeutic target for GC.
MicroRNAs/metabolism*
;
Humans
;
Stomach Neoplasms/pathology*
;
Cell Line, Tumor
;
Cell Movement/physiology*
;
Phosphatidylinositol 3-Kinases/metabolism*
;
Animals
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Mice
;
Octamer Transcription Factor-1/metabolism*
;
Mice, Nude
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Class Ia Phosphatidylinositol 3-Kinase/metabolism*
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Neoplasm Invasiveness
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Gene Expression Regulation, Neoplastic/genetics*
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Male
;
Immunohistochemistry
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Female
5.Atypical fibroxanthoma:clinicopathological features and prognostic analysis of 15 cases
Jiaying LIU ; Cui LIU ; Junhua WU ; Huizhen LI ; Xiu NIE ; Guixiang XIAO
Chinese Journal of Clinical and Experimental Pathology 2025;41(8):1044-1049
Purpose To investigate the clinicopathological features,differential diagnosis and prognosis of atypical fibroxanthoma(AFX).Methods Pathological features of 15 cases of AFX and 3 cases of pleomorphic dermal sarcoma(PDS)misdiagnosed as AFX were retrospectively analyzed by hematoxylin and eosin staining and immunohistochemical EnVision staining technology.Clinical information was collected and analyzed,and the relevant literatures were re-viewed.Results The age of the 15 patients with AFX ranged from 18 to 78 years,with an average age of 57 years.4 cases occurred in the head and neck,and 11 cases occurred in the trunk and limbs.3 patients with PDS misdiagnosed as AFX were aged from 56 to 60 years,with an average age of 58 years.The tumors were located in the trunk and limbs.Microscopically,15 cases of AFX and 3 cases of PDS misdiagnosed as AFX were composed of proliferative pleo-morphic and atypical spindle cells interspersed with a varying number of multinucleated cells.15 cases of AFX tumors were superficial and located in the dermis.In 3 cases of PDS misdiagnosed as AFX,1 case was located in subcutane-ous adipose tissue,1 case had superficial subcutaneous extension,and the third case had positive basal margin.Immu-nohistochemically,the immunophenotypes of the two groups were consistent.CD10 was expressed in all cases,CD68 was positive in most cases,SMA was expressed in a few cases,desmin was focal expressed in a very few cases,and S-100,SOX10,CD34,HMB-45,Melan A,STAT6 and CK(AE1/AE3)were not expressed in all cases.Ki67 prolifera-tion index ranged from 2%to 30%.15 patients with AFX were followed up from 12 to 108 months.One patient had tumor recurrence 1 year and 3 years after operation due to positive basal margin.Most of the other patients underwent extended resection after diagnosis and were in good condition without tumor recurrence and metastasis.3 cases of PDS misdiagnosed as AFX were followed up for 31 to 78 months.One patient had lung metastasis after 2 years,one patient recurred 4 times after operation,and the other patient died after 4 times of recurrence.Conclusion AFX is a rare dis-ease with similar pathological characteristics and immunophenotype to PDS.AFX can be diagnosed only when the tumor is small and completely confined to the dermis.When the maximum diameter of the tumor is more than 3 cm,or the presence of any form of subcutaneous extension requires a high level of vigilance for PDS.Careful differentiation and correct classification of AFX and PDS are very important for the treatment and prognosis of the disease.
6.Interleukin-13 is involved in vascular intimal hyperplasia by regulation of vascular smooth muscle phenotypic transformation
Xin WU ; Xiao LIU ; Jiaying ZHANG ; Ziyi ZHEN ; Qi LI ; Chang CHEN
Chinese Journal of Pathophysiology 2025;41(9):1694-1702
AIM:To investigate the mechanism by which interleukin-13(IL-13)influences vascular smooth muscle cell(VSMC)phenotypic transformation and subsequently contributes to vascular intimal hyperplasia in rats.METHODS:A total of 32 male SD rats,aged 5~7 weeks and weighing 330~360 g,were randomly divided into 4 groups(n=8 per group):normal(Nor)group,normal treatment(Nor+IL-13 neutralizing antibody,Nor+IL-13Nab)group,inju-ry(Inj)group,and injruy treatment(Inj+IL-13Nab)group.A 2F balloon catheter was used to induce mechanical injury in the left common carotid artery of SD rats to establish a vascular intimal hyperplasia model.Hematoxylin-eosin staining was performed to observe vascular structural changes.Enzyme-linked immunosorbent assay(ELISA)kits were used to measure IL-13 and transforming growth factor-β1(TGF-β1)levels.Human aortic smooth muscle cells(HA-SMCs)were cultured in vitro.Flow cytometry was conducted to assess peripheral blood CD4+IL-13+T cell content.Real-time quantita-tive PCR(RT-qPCR)was employed to evaluate gene expression levels of α-smooth muscle actin,osteopontin,calponin,collagen type Ⅰ/Ⅲ,proliferating cell nuclear antigen and Ki-67 antigen.Transwell and scratch wound assays were per-formed to assess cell migration.RESULTS:Compared with the model group,administration of IL-13Nab significantly in-hibited vascular intimal hyperplasia induced by mechanical vascular injury by antagonizing high plasma IL-13 levels(P<0.01).Immunofluorescence and mRNA analysis showed that neutralizing high plasma IL-13 suppressed collagen accumu-lation(P<0.01)and VSMC phenotypic transformation(P<0.01)in the injured vessels but did not inhibit peripheral blood CD4+IL-13+T cell activation.Incubation of HA-SMCs with recombinant human IL-13(rhIL-13)promoted cell pro-liferation and migration(P<0.01)as well as phenotypic transformation(P<0.01).Additional evidence suggested that rhIL-13-induced HA-SMC phenotypic transformation was associated with the regulation of TGF-β1 secretion by HA-SMCs.CONCLUSION:Interleukin-13 promotes vascular intimal hyperplasia by regulating VSMC phenotypic transformation through TGF-β1 secretion in rat models.
7.Atypical fibroxanthoma:clinicopathological features and prognostic analysis of 15 cases
Jiaying LIU ; Cui LIU ; Junhua WU ; Huizhen LI ; Xiu NIE ; Guixiang XIAO
Chinese Journal of Clinical and Experimental Pathology 2025;41(8):1044-1049
Purpose To investigate the clinicopathological features,differential diagnosis and prognosis of atypical fibroxanthoma(AFX).Methods Pathological features of 15 cases of AFX and 3 cases of pleomorphic dermal sarcoma(PDS)misdiagnosed as AFX were retrospectively analyzed by hematoxylin and eosin staining and immunohistochemical EnVision staining technology.Clinical information was collected and analyzed,and the relevant literatures were re-viewed.Results The age of the 15 patients with AFX ranged from 18 to 78 years,with an average age of 57 years.4 cases occurred in the head and neck,and 11 cases occurred in the trunk and limbs.3 patients with PDS misdiagnosed as AFX were aged from 56 to 60 years,with an average age of 58 years.The tumors were located in the trunk and limbs.Microscopically,15 cases of AFX and 3 cases of PDS misdiagnosed as AFX were composed of proliferative pleo-morphic and atypical spindle cells interspersed with a varying number of multinucleated cells.15 cases of AFX tumors were superficial and located in the dermis.In 3 cases of PDS misdiagnosed as AFX,1 case was located in subcutane-ous adipose tissue,1 case had superficial subcutaneous extension,and the third case had positive basal margin.Immu-nohistochemically,the immunophenotypes of the two groups were consistent.CD10 was expressed in all cases,CD68 was positive in most cases,SMA was expressed in a few cases,desmin was focal expressed in a very few cases,and S-100,SOX10,CD34,HMB-45,Melan A,STAT6 and CK(AE1/AE3)were not expressed in all cases.Ki67 prolifera-tion index ranged from 2%to 30%.15 patients with AFX were followed up from 12 to 108 months.One patient had tumor recurrence 1 year and 3 years after operation due to positive basal margin.Most of the other patients underwent extended resection after diagnosis and were in good condition without tumor recurrence and metastasis.3 cases of PDS misdiagnosed as AFX were followed up for 31 to 78 months.One patient had lung metastasis after 2 years,one patient recurred 4 times after operation,and the other patient died after 4 times of recurrence.Conclusion AFX is a rare dis-ease with similar pathological characteristics and immunophenotype to PDS.AFX can be diagnosed only when the tumor is small and completely confined to the dermis.When the maximum diameter of the tumor is more than 3 cm,or the presence of any form of subcutaneous extension requires a high level of vigilance for PDS.Careful differentiation and correct classification of AFX and PDS are very important for the treatment and prognosis of the disease.
8.Interleukin-13 is involved in vascular intimal hyperplasia by regulation of vascular smooth muscle phenotypic transformation
Xin WU ; Xiao LIU ; Jiaying ZHANG ; Ziyi ZHEN ; Qi LI ; Chang CHEN
Chinese Journal of Pathophysiology 2025;41(9):1694-1702
AIM:To investigate the mechanism by which interleukin-13(IL-13)influences vascular smooth muscle cell(VSMC)phenotypic transformation and subsequently contributes to vascular intimal hyperplasia in rats.METHODS:A total of 32 male SD rats,aged 5~7 weeks and weighing 330~360 g,were randomly divided into 4 groups(n=8 per group):normal(Nor)group,normal treatment(Nor+IL-13 neutralizing antibody,Nor+IL-13Nab)group,inju-ry(Inj)group,and injruy treatment(Inj+IL-13Nab)group.A 2F balloon catheter was used to induce mechanical injury in the left common carotid artery of SD rats to establish a vascular intimal hyperplasia model.Hematoxylin-eosin staining was performed to observe vascular structural changes.Enzyme-linked immunosorbent assay(ELISA)kits were used to measure IL-13 and transforming growth factor-β1(TGF-β1)levels.Human aortic smooth muscle cells(HA-SMCs)were cultured in vitro.Flow cytometry was conducted to assess peripheral blood CD4+IL-13+T cell content.Real-time quantita-tive PCR(RT-qPCR)was employed to evaluate gene expression levels of α-smooth muscle actin,osteopontin,calponin,collagen type Ⅰ/Ⅲ,proliferating cell nuclear antigen and Ki-67 antigen.Transwell and scratch wound assays were per-formed to assess cell migration.RESULTS:Compared with the model group,administration of IL-13Nab significantly in-hibited vascular intimal hyperplasia induced by mechanical vascular injury by antagonizing high plasma IL-13 levels(P<0.01).Immunofluorescence and mRNA analysis showed that neutralizing high plasma IL-13 suppressed collagen accumu-lation(P<0.01)and VSMC phenotypic transformation(P<0.01)in the injured vessels but did not inhibit peripheral blood CD4+IL-13+T cell activation.Incubation of HA-SMCs with recombinant human IL-13(rhIL-13)promoted cell pro-liferation and migration(P<0.01)as well as phenotypic transformation(P<0.01).Additional evidence suggested that rhIL-13-induced HA-SMC phenotypic transformation was associated with the regulation of TGF-β1 secretion by HA-SMCs.CONCLUSION:Interleukin-13 promotes vascular intimal hyperplasia by regulating VSMC phenotypic transformation through TGF-β1 secretion in rat models.
9.Study on the Interventional Effects of Cornus Officinalis Before and After Wine Preparation in Liu Wei Di Huang Decoction on Postmenopausal Osteoporosis Model Rats Based on Medicinal
Zhi WEN ; Hui YU ; Qiong LONG ; Jiaying TAN ; Li HUANG ; Wangzhong XIAO ; Bing DAI
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2717-2725
Objective To establish a dual wavelength HPLC method to compare the content changes of seven active components:monoglycoside,loganin,swertin,paeoniflorin,gallic acid,5-hydroxymethyl furfural and paeonol in Liuwei Dihuang Decoction(hereinafter referred to as"Liuweidihuang Decoction")before and after the preparation of Cornus wine;To explore the effect and mechanism of Liu(Shan)and Liu(Jiu)on 90 days after ovariectomy in postmenopausal osteoporosis model rats.Methods ①The contents of seven active ingredients in the Cornus wine mixed with Liuwei Dihuang Decoction were determined by dual wavelength HPLC.Chromatographic conditions:hypersil C18 chromatography(4.6 mm×250 mm,5 μm),mobile phase acetonitrile-0.3%phosphoric acid,gradient elution(0-7 min,0%-8%acetonitrile;7-15 min,8%-10%;15-20 min,10%-15%;20-30 min,15%-23%;30-50 min,23%-45%),flow rate 0.6 mL·min-1,detection wavelength:240 nm(monoglycoside,loganin,swertin,paeoniflorin),274 nm(gallic acid,5-hydroxymethyl furfural,paeonol),column temperature 30℃,injection volume 20 μL.②6-8 month old SD rats,divided into sham-operated group,model group,positive drug group,shanshuang flesh with Liuwei Dihuang Decoction group(liu(shan)group)and wine cornelian flesh withLiuwei Dihuang Decoction group(liu(wine)group),dosing started on the 5 d after ovariectomy,weighed weekly,and 5 rats from each group were taken at 90 d after surgery,weighed,anaesthetized,serum taken,bilateral femurs,and organs(uterus,spleen,liver,both kidneys and heart)were weighed.Serum calcium(Ca),phosphorus(P)and estradiol(E2)levels were measured using a fully automated biochemical instrument and chemiluminescent microparticle immunoassay,respectively;bone mineral density(BMD)values were measured using a dual-energy X-ray bone densitometer.HE staining was used to observe the morphological changes of bone tissue in rats.Results ①All the seven potent ingredients mentioned above were contained in the Liuwei Dihuang Decoction before and after the wine preparation of Cornu Cervi Pantotrichum,and the total content of the seven potent ingredients in liu(wine)was higher than that in liu(shan)(P<0.01).Among them,gallic acid and 5-hydroxymethylfurfural were higher in Liu(Jiu)than in Liu(Shan)(P<0.01),while the content of monosidine was lower than in Liu(Shan)(P<0.01).②Comparison with the sham-operated group:90 d after surgery,the serum Ca,P,E2 levels and uterine index of rats in the model group decreased(P<0.01),the BMD value decreased(P<0.01),and the degree of structural damage to bone tissue The degree of damage to the bone tissue structure was deepened,manifested by enlargement of the bone marrow cavity,gradual increase of adipocytes,dilution of the bone trabeculae arrangement,reduction of the number of bone trabeculae or even fracture.③Compared with the model group,90 d after surgery,the serum E2,Ca,P levels and uterine index of the rats in the liu(shan)and liu(wine)groups increased(P<0.05),the bone tissue structure of the liu(shan)and liu(wine)groups improved significantly,and the BMD value increased significantly(The liver and spleen indices of the liu(wine)groups increased(P<0.05).Conclusion The combination of the wine preparation of Cornus officinalis and its use in the formula of LiuWei Di Huang Decoction both contain monosidine,strychnine,sweretin,paeoniflorin,gallic acid,5-hydroxymethyl furfural(5-HMF)and tannin,and can effectively interfere with PMOP.Among them,the anti-osteoporotic effect of Liu Wei Di Huang Tang,which was formulated with wine and dogwood,was better than that of Liu Wei Di Huang Tang,which was formulated with Cornus officinalis,and the former contained more 5-HMF and gallic acid than the latter,which may provide some experimental basis for the selection of wine and dogwood in the 2020 edition of the Pharmacopoeia and the clinical application of Liu Wei Di Huang Tang in the prevention and treatment of PMOP.
10.Notch1 inhibits the mechanistic role of STING signaling to regulate hepatocyte lipophagy in nonalcoholic steatohepatitis
Tao YANG ; Jiaying ZHAO ; Xiao WANG ; Zegui FANG ; Longfeng JIANG ; Jun LI
Chinese Journal of Hepatology 2023;31(8):827-834
Objective:To study the mechanistic role of myeloid-specific Notch1 knockout inhibiting STING signaling to regulate hepatocyte lipophagy.Methods:A mouse model of nonalcoholic steatohepatitis (NASH) was established using a high-fat diet (HFD) and mouse bone marrow-derived macrophages (BMMs). Primary hepatocytes were isolated to construct a co-culture system. Twelve Notch1 FL/FL mice were randomly divided into two groups: the Notch1 FL/FL + normal diet (NCD) and the Notch1 FL/FL + HFD group. Further, 12 Notch1 M-KO mice were randomly divided into two groups: Notch1 M-KO + NCD, and Notch1 M-KO + HFD group.Serum alanine aminotransferase (sALT), total cholesterol (TC) and triglyceride (TG) were collected from mice serum samples. Liver tissue samples were collected for H&E staining, immunofluorescence (IF), Western blot and qRT-PCR. Tumor necrosis factor (TNF)-α was detected in the supernatant by enzyme-linked immunosorbent assay (ELISA). The comparison of inter group data was conducted using a t-test. Results:The mouse NASH model, mouse BMMs co-culture system, and primary hepatocytes were successfully constructed. Compared with the Notch1 FL/FL + HFD group, the Notch1 M-KO + HFD group showed a significant increase in serum ALT [(250.02 ± 58.21) U/L vs (370.70 ± 54.57) U/L, t = 3.705, P = 0.004], TG [(29.90 ± 3.54) mg/g vs (43.83 ± 8.56) mg/g, t = 3.685, P = 0.004], and TC [(33.70 ± 8.43) mg/g vs (90.53 ± 12.53) mg/g, t = 9.917, P < 0.001]. HE staining of liver tissue showed remarkable balloon-like alterations in liver cells, while IF staining demonstrated increased macrophage infiltration ( t = 7.346, P < 0.001). Compared with the hepatocyte group co-cultured with Notch1 FL/FL BMMs, the BODIPY probe showed a significant increase in lipid droplet (LDs) deposition in liver cells in the Notch1 M-KO group ( t = 3.835, P < 0.001). The co-localization of lysosomal associated membrane protein 1 (LAMP1), LDs ( t = 7.103, P < 0.001), microtubule-associated protein light chain 3 (LC3) -II/LC3-I ( t = 5.0, P = 0.007), and autophagy associated gene 12 (Atg12) ( t = 28.36, P < 0.001) had decreased expression, while P-62 had increased expression ( t = 3.253, P = 0.03), indicating a decrease in autophagic flow. Additionally, LC3 and LDs colocalization decreased ( t = 5.24, P = 0.0003), indicating reduced lipophagy. Compared with the Notch1 FL/FL group, the Notch1 M-KO BMMS mouse group showed an increase in the expression of p-STING ( t = 5.318, P = 0.006), p-TANK1 binding kinase 1 (TKB1) ( t = 6.467, P = 0.002), p-interferon regulatory factor 3 (IRF3) ( t = 14.61, P < 0.001), and p-P65 ( t = 12.7, P = 0.002) protein, accompanied by mRNA expression of the inflammatory mediators interferon (IFN)-β ( t = 7.978, P < 0.001), TNFα ( t = 8.496, P = 0.001), interleukin-1 β (IL-1 β) ( t = 4.7, P < 0.001), and CXCL-10 ( t = 4.428, P = 0.001). The STING gene was knocked out in the BMMs Notch1 M-KO mice using CRISPR/Cas9. Compared with the CRISPR-Control group, the expression of P-TKB1 ( t = 2.909, P = 0.044), p-IRF3 ( t = 10.96, P < 0.001), p-IRF3 ( t = 10.96, P < 0.001), and p-P65 ( t = 7.091, P = 0.002) proteins was lower in the STING-KO BMMs group. The release of TNF-α in the supernatant was decreased (732.3 ± 129.35 pg/ml vs. 398.17 ± 47.15 pg/ml, t = 4.204, P = 0.014). However, in hepatocytes co-cultured with STING-KO BMMs, LC3-II/LC3-I ( t = 7.546, P = 0.001) increased, p-62 ( t = 10.96, P < 0.001) expression decreased, autophagic flow increased, and the colocalization of LC3 and LDs increased, lipophagy increased, and LDs deposition decreased. Conclusion:Myeloid-specific Notch1 knockout can activate macrophages STING signaling, increase the expression of inflammatory mediator genes, inhibit the occurrence of autophagy flow and lipophagy in hepatocyte cells, and aggravate LDs deposition and NASH progression.

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