1.Mechanism of mesenchymal stromal cells reversing fibrosis of the uterine endometrium after injury
Shiqi LI ; Yue DANG ; Qingqing ZHOU ; Xin SHANG ; Jiali LIU
Journal of China Pharmaceutical University 2026;57(3):360-368
To investigate the effect of mesenchymal stromal cells (MSCs) on the fibrosis progression and endometrial regeneration in mouse models of endometrial injury, a mouse endometrial injury model was established by physical scraping combined with lipopolysaccharide (LPS) chemical induction, and the endometrium morphology and collagen deposition were evaluated by HE and Masson staining after unilateral administration of MSCs. RT-qPCR and immunofluorescence were used to detect the expression of epithelial markers Epcam and collagen I (Col1). Primary endometrial epithelial cells were isolated to assess p21 expression. Fibrosis-related genes and signaling pathways were validated using transcriptome sequencing and RT-qPCR. The experimental results showed that MSCs increased endometrial thickness and number of glands (P<0.05), up-regulated the expression of Epcam (P<0.01), while significantly reducing uterine collagen fiber deposition (P<0.05) and the expression of p21 in primary epithelial cells (P<0.0001). Transcriptomic analysis revealed that MSCs attenuated collagen deposition by modulating genes involved in the TGF-β signaling pathway. This study elucidates that MSCs facilitate uterine repair by reversing endometrial fibrosis and promoting epithelial regeneration, and their role is related to the regulation of the TGF-β/SMAD pathway and suppression of aberrant collagen accumulation.
2.Predictive value of the early 18F-fluorodeoxyglucose positron emission tomography/computed tomography parameters percentage change in the maximum standardized uptake value and metabolic tumor volume for residual tumor lesions in patients with diffuse large B-cell lymphoma after chemotherapy
Chunjiang DANG ; Qian YU ; Fangqi ZHOU ; Lin FANG
Chinese Journal of Radiological Health 2026;35(3):414-420
Objective To analyze the changes of early 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET-CT) metabolic parameters in patients with diffuse large B-cell lymphoma (DLBCL) before and after chemotherapy, and to systematically evaluate the early predictive efficacy of the above prarmeters for residual lesions. Methods Eighty patients with DLBCL admitted to our hospital from January 2021 to January 2025 were randomly selected as study subjects. All patients received 18F-FDG PET-CT examination. After 3 months of chemotherapy, patients were assessed according to the Lugano 2014 criteria. Patients without residual lesions were assigned to the complete remission group, while those with residual lesions were assigned to the residual lesion group. Clinical data and 18F-FDG PET-CT parameters were compared between the two groups. A logistic regression model was used to analyze the relationship between 18F-FDG PET-CT parameters and residual tumor lesions in DLBCL patients after chemotherapy. The receiver operating characteristic curve was used to analyze the predictive value of early 18F-FDG PET-CT parameters for residual tumor lesions in patients with DLBCL after chemotherapy. Results Compared with the complete remission group, the residual lesion group had a significantly higher proportion of patients with an international prognostic index score>3, as well as significantly higher maximum standardized uptake value (SUVmax), metabolic tumor volume (MTV), and total lesion glycolysis(TLG). The early percentage change in the maximum standardized uptake value (ΔSUVmax%) was significantly lower in the residual lesion group (P<0.05). There were no significant differences in age, sex, Ann Arbor stage, serum lactate dehydrogenase elevation, Eastern Cooperative Oncology Group performance status score ≥2, and molecular subtype between the two groups (P>0.05). Logistic regression analysis showed that an international prognostic index score >3 and a high MTV were independent risk factors for residual tumor lesions in DLBCL patients after chemotherapy, while a high ΔSUVmax% was a protective factor (P<0.05). Receiver operating characteristic curve analysis showed that the area under the curve for ΔSUVmax% and MTV alone in predicting residual tumor lesions after chemotherapy for DLBCL was 0.918 (95%CI=0.848-0.989) and 0.926 (95%CI=0.866-0.985), with cutoff values of 75.16% and 13.58 cm3, respectively. The combined prediction using both parameters yielded an area under the curve of 0.960 (95%CI=0.891-0.991), with a sensitivity of 91.38% and a specificity of 95.45%. Conclusion The early 18F-FDG PET-CT parameters ΔSUVmax% and MTV demonstrate high predictive value for residual tumor lesions in DLBCL patients after chemotherapy. Their combined use has improved predictive efficacy, which can provide a reference for clinical evaluation of chemotherapy efficacy in DLBCL.
3.Analysis of factors influencing early recurrence for patients with initially unresectable hepa-tocellular carcinoma who underwent liver resection following downstaging treatment and construction of a predictive model: a multicenter study
Yun YANG ; Peng LU ; Kongying LIN ; Zheng DANG ; Wei GUO ; Zeya PAN ; Weiping ZHOU
Chinese Journal of Digestive Surgery 2025;24(2):223-235
Objective:To investigate the factors influencing early recurrence for patients with initially unresectable hepatocellular carcinoma (HCC) who underwent salvage liver resection (SLR) following transcatheter arterial chemoembolization-based downstaging treatment, and construct a predictive model to evaluate its predicting performance.Methods:The retrospective cohort study was constructed. The clinicopathological data of 305 patients with initially unresectable HCC who were admitted to 4 medical centers in China, including the Third Affiliated Hospital of Naval Medical University (Shanghai Eastern Hepatobiliary Surgery Hospital) et al, from January 2019 to December 2021 were collected. There were 286 males and 19 females, aged (48.7±10.4)years. A total of 133 patients who were admitted from January 2019 to December 2020 were set as the training cohort, and the other 172 patients who were admitted from January to December 2021 were set as the validation cohort. Observation indicators: (1) postoperative recurrence-free survival in HCC patients; (2) analysis of factors influencing postoperative early recurrence in HCC patients; (3) construction and validation of the predictive model. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data was conducted using the rank sum test. Univariate and multivariate analyses were conducted using the Cox regre-ssion model. The Kaplan-Meier method was used to calculate survival. The Log-rank test was used for survival analysis. The predicting performance of the model was evaluated using the concordance index (C-index) and the area under curve (AUC) of time-dependent receiver operating characteristic (ROC) curve, and the accuracy of the model was validated using the calibration curve. The total net gain of the model was evaluated using the decision curve. Results:(1) Postoperative recurrence-free survival in HCC patients. The recurrence-free survival time of 133 HCC patients in the training cohort was 10.0(range, 1.5-24.0)months, with 1-, 2-year recurrence-free survival rate of 47.3% and 36.8%. The recurrence-free survival time of 172 HCC patients in the validation cohort was 11.0(range, 1.0-24.0)months, with 1-, 2-year recurrence-free survival rate of 51.7% and 37.2%. There was no significant difference in recurrence-free survival between patients in the training cohort and the validation cohort ( χ2=0.075, P>0.05). (2) Analysis of factors influencing postoperative early recur-rence in HCC patients. Results of multivariate analysis showed that tumor burden prior to down-staging treatment, grade of albumin-bilirubin (ALBI) score prior to SLR, alpha-fetoprotein (AFP) half-life prior to SLR, and tumor response prior to SLR were independent factors influencing early recurrence in HCC patients after surgery [ hazard ratio=3.212, 2.526, 2.304, 1.575, 95% confidence interal ( CI) as 1.262-8.175, 1.324-4.818, 1.477-3.595, 1.138-2.180, P<0.05]. (3) Construction and validation of the predictive model. A nomogram predictive model for postoperative early recurrence was constructed base on the results of multivariate analysis. The C-index of predictive model was 0.786 for the training cohort and 0.734 for the validation cohort. The AUC of ROC curve of nomogram predictive model for 12-, 18-, and 24-month recurrence-free survival rate in the training cohort were 0.890 (95% CI as 0.836-0.944), 0.895 (95% CI as 0.842-0.947), and 0.887 (95% CI as 0.831-0.942), respectively. The AUC of ROC curve of nomogram predictive model for 12-, 18-, and 24-month recurrence-free survival rate in the validation cohort were 0.845 (95% CI as 0.781-0.909], 0.888 (95% CI as 0.826-0.950), and 0.919 (95% CI as 0.870-0.968), respectively. Results of calibration curve showed high consistency between the predicted results of nomogram predictive model and actual outcomes. Results of decision curve showed the nomogram predictive model with a good total net gain at a threshold of 0.10-0.50. Conclusions:Tumor burden prior to downstaging treatment, grade of ALBI score prior to SLR, AFP half-life prior to SLR, and tumor response prior to SLR are independent factors influencing early recurrence in initially unresectable HCC patients undergoing SLR following downstaging treatment. The nomogram predictive model based on these factors can effectively evaluate the prognosis of this patient population.
4.Clinical value of evaluation of anatomical position of inferior mesenteric artery and vein as well as left colic artery in laparoscopic radical resection of colorectal
Bo ZHANG ; Tao WU ; Shuai ZHOU ; Ling DANG ; Jiaxing HE ; Ying YANG ; Zhuo HAN ; Longlong ZHENG ; Nan WANG
Chinese Journal of Digestive Surgery 2025;24(6):754-761
Objective:To investigate the clinical value of laparoscopic evaluation of anato-mical position of inferior mesenteric artery (IMA), inferior mesenteric vein (IMV) and left colic artery (LCA).Methods:The prospective one-arm study was conducted. The clinical data of 229 pati-ents who underwent laparoscopic left hemicolectomy for left colon or laparoscopic radical resection of rectal cancer in The Second Affiliated Hospital of Air Force Medical University from December 2022 to December 2023 were selected. The distance between the origin point of IMA and the origin point of the first branch (L1) as well as the distance from the origin point of LCA root to the junction of LCA and IMV (L2) were measured during the operation. IMA classification, the location relation-ship of LCA and IMV junction were recorded. Observation indicators: (1) situations of enrolled patients; (2) difference analysis between L1, L2 and clinical features; (3) distribution characteristics of the location relationship between LCA and IMV in different types of IMA. Mann-Whitney U test was used for comparison of measurement data with skewed distribution between groups, Kruskal-Wallis H test was used for comparison between multiple groups, and Dunn-Bonferroni test was used for pairwise comparison. Comparison of count data between groups was performed by chi-square test. Pearson or Spearman correlation analysis was conducted for correlation of continuous variables. Results:(1) Situations of enrolled patients. A total of 229 eligible patients were screened out, including 146 males and 83 females, aged 64(range, 55-71)years. The height of 229 patients was 168(range, 160-172)cm, the weight was 65.0(55.5,71.5)kg, the body surface area was (1.68±0.17)m 2, the tumor maximum diameter was 3.0(2.5,4.0)cm. The total number of lymph nodes dissected was 19(17,21), and the number of No.253 lymph node dissected was 4(3,5). The L1 was 3.50(1.20,8.00)cm, and the L2 was 2.20(0.50,7.30)cm. There were 58, 31, 32, 71, 22, 90, 26 and 212 patients with smoking, alcohol drinking, diabetes, hypertension, coronary heart disease, neoadjuvant chemo-therapy, neoadjuvant radiotherapy and preservation of the LCA, respectively. Among 229 patients, cases with BMI <18.5 kg/m 2, 18.5-23.9 kg/m 2 and >23.9 kg/m 2 were 11, 133 and 85, respectively. There were 153 cases in pathological stage Ⅰ-Ⅱ and 76 cases in stage Ⅲ. There were 168 cases of Dixon operation, 6 cases of Miles operation and 55 cases of sigmoid colon resection. There were 135 cases of IMA type 1, 44 cases of IMA type 2, 23 cases of IMA type 3, 2 cases of IMA type 4, and 25 cases of IMA type unable to judge. (2) Difference analysis between L1, L2 and clinical features.Correlation analysis showed negative correlation between the height, body surface area and L1 ( r=-0.17, -0.15, P<0.05). The L1 was 3.20(2.68,4.00)cm for male patients and 3.60(3.00,4.20)cm for female patients, respectively, showing a significant difference between the two groups ( Z=-2.37, P<0.05). The L1 of patients with IMA type 1, 2, and 3 was 3.20(2.80,4.00)cm, 3.85(3.00,4.48)cm, and 3.20(2.50,4.30)cm, respectively, showing a significant difference among them ( H=7.54, P<0.05). Further pairwise com-parison showed that there was a significant difference in L1 between patients with IMA type 2 and those with IMA type 1 ( P<0.05). The L2 of smokers and non-smokers were 2.50(1.95,3.20)cm and 2.20(1.60,2.80)cm, respectively, showing a significant difference between the two groups ( Z=-2.24, P<0.05). (3)Distribution characteristics of the location relationship between LCA and IMV in different types of IMA. There was no significant difference in LCA distribution between the anterior and posterior positions of IMV among the three IMA types (type 1, 2, 3) ( χ2=1.63, P>0.05). Conclusions:Patients with greater height have larger body surface area and shorter L1. L1 is significantly longer in female patients than in male patients. L1 is significantly longer in patients with IMA type 2 than in those with type 1. L2 is significantly longer in smokers than in non-smokers. There was no significant difference in the distribution location between LCA and IMV among patients of IMA type 1, 2 and 3.
5.Individualized dosage study of vitamin D3 based on physiologically-based pharmacokinetic modeling
Yuanyuan WEI ; Tao MA ; Yuezhou TANG ; Hubo LI ; Xiaoyu TIAN ; Yunjie DANG ; Xu ZHOU
Chinese Journal of Clinical Pharmacology and Therapeutics 2025;30(8):1067-1075
AIM:To establish a physiologically-based pharmacokinetic(PBPK)model for vitamin D in adults,aiming to provide guidance for the ratio-nal clinical use of vitamin D in individuals with vita-min D deficiency.METHODS:Relevant literature and databases were reviewed to obtain the physi-cochemical properties and pharmacokinetic param-eters of vitamin D3.The PBPK model for adult whole-body vitamin D was constructed,optimized,and predicted using PK-Sim? software.The model's predictive performance was evaluated using confi-dence intervals,goodness of fit,and fold error(FE).The effectiveness of commonly used clinical dosing regimens was assessed based on the final opti-mized model,and personalized dosing recommen-dations were provided.RESULTS:The established adult whole-body PBPK model for vitamin D had a goodness of fit R2 of 0.961,approaching 1,and the FE values for AUC0-∞ and Cmax were both within the range of 0.5 and 2,indicating that the constructed PBPK model possesses good data predictive capa-bility.CONCLUSION:A successful PBPK model for oral vitamin D3 in adults has been established,showing good predictive performance for single oral doses of vitamin D3.Single oral doses of vita-min D3(7 500 μg and 15 000 μg)are safe and effec-tive dosing regimens for improving vitamin D insuf-ficiency or deficiency in Asian adults.Regular moni-toring of vitamin D levels before and during treat-ment is recommended to achieve the optimal out-comes of personalized therapy.
6.Association between TMED4 and the Immune Microenvironment in Colorectal Cancer
Dang ZHOU ; Junrong LI ; Qi LIU
Journal of Medical Research 2025;54(6):127-132
Objective To explore the clinical significance of TMED4 expression in colorectal cancer and its relationship with the im-mune microenvironment.Methods Based on the cancer genome atlas(TCGA)database,the expression level of TMED4 in colorectal cancer and its prognostic value were analyzed using R language.Clinical samples were collected for polymerase chain reaction(PCR)ex-periments to verify its expression,and the correlation with clinicopathological features and prognostic value were analyzed.The functional role of TMED4 was detected through gene set enrichment analysis(GSEA)and immune infiltration analysis.Results The expression level of TMED4 was upregulated in colorectal cancer samples.The expression level of TMED4 was closely associated with the pathological stage,lymphatic metastasis,and distant metastasis of colorectal cancer.The increase in TMED4 expression was closely related to the over-all survival(OS)and disease-specific survival(DSS)of patients.Univariate analysis and multivariate COX analysis found that TMED4might be an independent high-risk factor for DSS in colorectal cancer patients.Receiver operating characteristic(ROC)curve a-nalysis showed that TMED4has the potential to become a potential biomarker for the diagnosis of colorectal cancer.GSEA revealed that TMED4might be involved in multiple pathways,mainly concentrated in transport and catabolic pathways,glycan biosynthesis and metabo-lism pathways,and lipid metabolism pathways.Conclusion High expression of TMED4 is closely related to poor prognosis of colorectal cancer and immune cell infiltration.It may serve as an independent biomarker for predicting the prognosis of colorectal cancer and a poten-tial therapeutic target.
7.Analysis of influencing factors and construction of predictive models of immune-related skin adverse events in urologic neoplasms
Ran SUN ; Kai DANG ; Yongan ZHOU ; Yang YANG ; Xiangyu WANG ; Jinhua LIU ; Jing XIAO ; Teng CUI
International Journal of Surgery 2025;52(10):665-671
Objective:To investigate the risk factors of skin adverse events associated with immune checkpoint inhibitor (ICI) therapy in patients with urologic neoplasms, and establish a predictive model.Methods:A single-center retrospective case-control study enrolled 91 advanced urologic neoplasms patients who received ICI therapy at the Department of Urology, Beijing Friendship Hospital, Capital Medical University from January 2020 to June 2025. Patients were divided into the skin lesion group ( n=44) and the control group ( n=47). Patients in the skin lesion group experienced related skin adverse events during ICI treatment, while patients in the control group did not experience such events during ICI treatment. The general data and laboratory indicators were compared between the two groups. The normally distributed measurement data were expressed as mean±standard deviation ( ± s), and the independent sample t-test was used for comparison between groups; the non-normally distributed measurement data were expressed as the median (interquartile range) [ M ( Q1, Q3)], and the Kruskal-Wallis test was used for comparison between groups. The count data were expressed as the number of cases and percentages, and the Chi-test was used for comparison between groups. First, a univariate analysis was conducted on the influencing factors of skin adverse events in patients with urologic neoplasms after ICI treatment. Then, the indicators with statistically significant differences in the univariate analysis were further included in the multivariate Logistic regression model to screen the independent risk factors for predicting skin adverse events. The R software was used to incorporate the factors with significant differences from multivariate analysis into the prediction model and construct a Nomogram. The calibration curve was utilized to evaluate the consistency between predicted values and actual observed results. Meanwhile, the discrimination of the model was verified by the receiver operating characteristic (ROC) curve and the area under the curve (AUC), so as to comprehensively verify the reliability and clinical application value of the prediction model. Results:The results of univariate analysis showed that there were statistically significant differences between the skin lesion group and the control group in terms of the proportion of other immune responses, serum albumin level, absolute eosinophil count, and C-reactive protein (CRP) levels ( P<0.05). These factors were included in multivariate Logistic regression, which identified elevated absolute eosinophil count and elevated CRP as the independent risk factors for related skin adverse events in patients with urologic neoplasms after ICI treatment. A predictive nomogram was built based on these factors. The calibration curve showed high consistency between predicted and actual probabilities, and ROC analysis confirmed the combined model had high predictive value (AUC=0.883, P<0.001). Conclusions:Elevated absolute eosinophil count and elevated CRP level are independent predictors of immune-related skin adverse events in urologic neoplasms patients after ICI treatment. The prediction model constructed based on these two factors facilitates early clinical screening and identification of high-risk patients.
8.A case report of neurodevelopmental disorder caused by mutation of the RAB11B gene
Xi ZHANG ; Xiubo DU ; Zhiru WANG ; Huawei LI ; Weili DANG ; Yuxiang YE ; Rongyi ZHOU
Chinese Journal of Neurology 2025;58(2):184-187
The purpose of this investigation was to elucidate the clinical characteristics and genetic underpinnings of a pediatric patient with neurodevelopmental disorder with ataxic gait, absent speech, and decreased cortical white matter (NDAGSW, OMIM#617807). The affected individual, a 1-year-9-month-old male, displayed physical development retardation and distinctive facial features, notably periorbital puffiness, upward-gazing palpebral fissures, a shortened philtrum, a tented mouth, and conical-shaped digits. Clinically, the patient presented with profound global developmental retardation, marked language deficits, hypotonia, and an ataxic gait. Subtle, non-diagnostic alterations were identified in cranial magnetic resonance imaging and visual evoked potential assessments. The trio-whole exome sequencing analysis revealed a de novo heterozygous mutation, c.202G>A (p.A68T), within the RAB11B gene, a known pathogenic variant linked to NDAGSW. Neurodevelopmental disorders due to RAB11B gene variants are rare disorders with clinical manifestations of severe mental retardation, aphasia, motor retardation, gait abnormalities with peculiar phenotypical features, structural abnormalities of the brain, and reduced cerebral white matter, cerebellar hypoplasia, and hypoplasia of the corpus callosum as seen on cranial imaging. Based on the characteristics of the disease, the heterozygous missense mutation c.202G>A (p.Ala68Thr) in the RAB11B gene was identified as the genetic etiology of the child.
9.Modified Lianpoyin Formula Treats Hp-associated Gastritis by Regulating Mitochondrial Autophagy and NLRP3 Inflammasome Signaling Pathway
Siyi ZHANG ; Haopeng DANG ; Wenliang LYU ; Wentao ZHOU ; Wei GUO ; Lin LIU ; Lan ZENG ; Yujie SUN ; Luming LIANG ; Yi ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):178-187
ObjectiveTo explore the effect of modified Lianpoyin formula (LPYJWF) in the treatment of Helicobacter pylori (Hp)-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control, model, high-dose LPYJWF (LPYJWF-H, 27.3 g·kg-1·d-1), medium-dose LPYJWF (LPYJWF-M, 13.65 g·kg-1·d-1), low-dose LPYJWF (LPYJWF-L, 6.83 g·kg-1·d-1), and quadruple therapy groups. Except the control group, other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole (6.06 mg·kg-1·d-1) + amoxicillin (303 mg·kg-1·d-1) + clarithromycin (151.67 mg·kg-1·d-1) + colloidal pectin capsules (30.3 mg·kg-1·d-1) by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of gastric mucosa, and Warthin-Starry (W-S) silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue, and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A (TFAM) and translocase of the outer mitochondrial membrane member 20 (TOMM20). The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase (SOD) and malondialdehyde (MDA), respectively, in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 (PINK1), Parkin, p62, microtubule-associated protein 1 light chain 3 (LC3), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), interleukin-1β (IL-1β), and interleukin-18 (IL-18). Real-time quantitative PCR was employed to assess the mRNA levels of PINK1, Parkin, p62, and LC3. ResultsCompared with the control group, the model group presented obvious gastric mucosal damage, colonization of a large number of Hp, severe mitochondrial damage, vacuolated structures due to excessive autophagy, reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue, and reduced SOD and increased MDA (P<0.01). In addition, the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1, Parkin, and LC3 and down-regulated protein and mRNA levels of p62 (P<0.01, as well as increased expression of inflammasome-associated proteins NLRP3, ASC, IL-1β, and IL-18 (P<0.01). Compared with the model group, the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa, reduced Hp colonization, mitigated mitochondrial damage, and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group (P<0.01), and the MDA levels became lowered in the LPYJWF and quadruple therapy groups (P<0.05, P<0.01). Furthermore, the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1, Parkin, and LC3 and protein levels of PINK1 and Parkin, and up-regulated mRNA level of p62 (P<0.01). The LPYJWF-M, LPYJWF-H, and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level (P<0.05, P<0.01) and up-regulated protein level of p62 (P<0.01). The expression of inflammasome-associated proteins NLRP3, ASC, IL-1β, and IL-18 were reduced in the LPYJWF and quadruple therapy groups (P<0.05, P<0.01). ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice, which may be related to the inhibition of excessive mitochondrial autophagy, thereby inhibiting the activation of the NLRP3 inflammasome pathway.
10.Modified Lianpoyin Formula Treats Hp-associated Gastritis by Regulating Mitochondrial Autophagy and NLRP3 Inflammasome Signaling Pathway
Siyi ZHANG ; Haopeng DANG ; Wenliang LYU ; Wentao ZHOU ; Wei GUO ; Lin LIU ; Lan ZENG ; Yujie SUN ; Luming LIANG ; Yi ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):178-187
ObjectiveTo explore the effect of modified Lianpoyin formula (LPYJWF) in the treatment of Helicobacter pylori (Hp)-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control, model, high-dose LPYJWF (LPYJWF-H, 27.3 g·kg-1·d-1), medium-dose LPYJWF (LPYJWF-M, 13.65 g·kg-1·d-1), low-dose LPYJWF (LPYJWF-L, 6.83 g·kg-1·d-1), and quadruple therapy groups. Except the control group, other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole (6.06 mg·kg-1·d-1) + amoxicillin (303 mg·kg-1·d-1) + clarithromycin (151.67 mg·kg-1·d-1) + colloidal pectin capsules (30.3 mg·kg-1·d-1) by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of gastric mucosa, and Warthin-Starry (W-S) silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue, and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A (TFAM) and translocase of the outer mitochondrial membrane member 20 (TOMM20). The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase (SOD) and malondialdehyde (MDA), respectively, in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 (PINK1), Parkin, p62, microtubule-associated protein 1 light chain 3 (LC3), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), interleukin-1β (IL-1β), and interleukin-18 (IL-18). Real-time quantitative PCR was employed to assess the mRNA levels of PINK1, Parkin, p62, and LC3. ResultsCompared with the control group, the model group presented obvious gastric mucosal damage, colonization of a large number of Hp, severe mitochondrial damage, vacuolated structures due to excessive autophagy, reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue, and reduced SOD and increased MDA (P<0.01). In addition, the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1, Parkin, and LC3 and down-regulated protein and mRNA levels of p62 (P<0.01, as well as increased expression of inflammasome-associated proteins NLRP3, ASC, IL-1β, and IL-18 (P<0.01). Compared with the model group, the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa, reduced Hp colonization, mitigated mitochondrial damage, and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group (P<0.01), and the MDA levels became lowered in the LPYJWF and quadruple therapy groups (P<0.05, P<0.01). Furthermore, the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1, Parkin, and LC3 and protein levels of PINK1 and Parkin, and up-regulated mRNA level of p62 (P<0.01). The LPYJWF-M, LPYJWF-H, and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level (P<0.05, P<0.01) and up-regulated protein level of p62 (P<0.01). The expression of inflammasome-associated proteins NLRP3, ASC, IL-1β, and IL-18 were reduced in the LPYJWF and quadruple therapy groups (P<0.05, P<0.01). ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice, which may be related to the inhibition of excessive mitochondrial autophagy, thereby inhibiting the activation of the NLRP3 inflammasome pathway.

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