1.Mechanism of pachymic acid in ameliorating renal injury in pregnancy induced hypertension rats by regulating the Sirt1/PGC‑1α pathway
Junjiang ZHU ; Jincheng LIN ; Jiajian WU ; Yi ZENG ; Jun HU ; Min LI ; Hongying LIU ; Jinfen LI
China Pharmacy 2026;37(2):186-191
OBJECTIVE To investigate the mechanism of pachymic acid on renal injury in pregnancy induced hypertension (PIH) rats by regulating the silent information regulator transcript 1/peroxisome proliferator-activated receptor γ coactivator-1α (Sirt1/PGC-1α) pathway. METHODS Pregnant SD rats were prepared by co-caging and PIH model was induced using N-nitro-L- arginine methyl ester (L-NAME) method. PIH rats were randomly divided into model group, L-pachymic acid (low-dose pachymic acid, 10 mg/kg) group, H-pachymic acid (high-dose pachymic acid, 20 mg/kg) group, and H-pachymic acid+EX527 (20 mg/kg pachymic acid+10 mg/kg EX527) group, with 6 rats in each group. Another 6 normal pregnant rats were selected as blank group. Each group was given relevant medicine or solvent intragastrically or intraperitoneally daily, once a day, for 28 consecutive days. After the last administration, 24 h urinary protein and tail artery systolic blood pressure (SBP) were measured in pregnant rats from each group, along with the levels of serum creatinine (Scr), blood urea nitrogen (BUN),uric acid (UA), and cystatin C (Cys-C). The contents of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in renal tissue, as well as the mRNA and protein expression levels of Sirt1 and PGC-1α, were also determined. Meanwhile, renal histopathological changes in rats from each group were evaluated using hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining. RESULTS Compared with model group, L-pachymic acid group and H-pachymic acid group exhibited significant decreases in 24 h urine protein quantification, tail artery SBP, Scr, BUN, UA, Cys-C levels, glomerulosclerosis index score of renal tissue, renal tubular injury score, the percentage of PAS positive area, MDA and 8-OHdG (P<0.05). Conversely, the contents of SOD and GSH-Px, along with the mRNA and protein expression levels of Sirt1 and PGC-1α, were significantly increased (P<0.05). Moreover, these improvements were more pronounced in H-pachymic acid group (P<0.05). Compared with H-pachymic acid group, the aforementioned indicators in pregnant rats from the H-pachymic acid+EX527 group showed significant reversal (P<0.05). CONCLUSIONS Pachymic acid significantly ameliorates renal injury induced by PIH in rats, potentially through activation of the Sirt1/PGC-1α pathway.
2.The Use of Speech in Screening for Cognitive Decline in Older Adults
Si-Wen WANG ; Xiao-Xiao YIN ; Lin-Lin GAO ; Wen-Jun GUI ; Qiao-Xia HU ; Qiong LOU ; Qin-Wen WANG
Progress in Biochemistry and Biophysics 2025;52(2):456-463
Alzheimer’s disease (AD) is a chronic neurodegenerative disorder that severely affects the health of the elderly, marked by its incurability, high prevalence, and extended latency period. The current approach to AD prevention and treatment emphasizes early detection and intervention, particularly during the pre-AD stage of mild cognitive impairment (MCI), which provides an optimal “window of opportunity” for intervention. Clinical detection methods for MCI, such as cerebrospinal fluid monitoring, genetic testing, and imaging diagnostics, are invasive and costly, limiting their broad clinical application. Speech, as a vital cognitive output, offers a new perspective and tool for computer-assisted analysis and screening of cognitive decline. This is because elderly individuals with cognitive decline exhibit distinct characteristics in semantic and audio information, such as reduced lexical richness, decreased speech coherence and conciseness, and declines in speech rate, voice rhythm, and hesitation rates. The objective presence of these semantic and audio characteristics lays the groundwork for computer-based screening of cognitive decline. Speech information is primarily sourced from databases or collected through tasks involving spontaneous speech, semantic fluency, and reading, followed by analysis using computer models. Spontaneous language tasks include dialogues/interviews, event descriptions, narrative recall, and picture descriptions. Semantic fluency tasks assess controlled retrieval of vocabulary items, requiring participants to extract information at the word level during lexical search. Reading tasks involve participants reading a passage aloud. Summarizing past research, the speech characteristics of the elderly can be divided into two major categories: semantic information and audio information. Semantic information focuses on the meaning of speech across different tasks, highlighting differences in vocabulary and text content in cognitive impairment. Overall, discourse pragmatic disorders in AD can be studied along three dimensions: cohesion, coherence, and conciseness. Cohesion mainly examines the use of vocabulary by participants, with a reduction in the use of nouns, pronouns, verbs, and adjectives in AD patients. Coherence assesses the ability of participants to maintain topics, with a decrease in the number of subordinate clauses in AD patients. Conciseness evaluates the information density of participants, with AD patients producing shorter texts with less information compared to normal elderly individuals. Audio information focuses on acoustic features that are difficult for the human ear to detect. There is a significant degradation in temporal parameters in the later stages of cognitive impairment; AD patients require more time to read the same paragraph, have longer vocalization times, and produce more pauses or silent parts in their spontaneous speech signals compared to normal individuals. Researchers have extracted audio and speech features, developing independent systems for each set of features, achieving an accuracy rate of 82% for both, which increases to 86% when both types of features are combined, demonstrating the advantage of integrating audio and speech information. Currently, deep learning and machine learning are the main methods used for information analysis. The overall diagnostic accuracy rate for AD exceeds 80%, and the diagnostic accuracy rate for MCI also exceeds 80%, indicating significant potential. Deep learning techniques require substantial data support, necessitating future expansion of database scale and continuous algorithm upgrades to transition from laboratory research to practical product implementation.
3.Comprehensive evaluation of benign and malignant pulmonary nodules using combined biological testing and imaging assessment in 1 017 patients: A retrospective cohort study
Lei ZHANG ; Zihao LI ; Nan LI ; Jun CHENG ; Feng ZHANG ; Pinghui XIA ; Wang LÜ ; ; Jian HU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(01):60-66
Objective By combining biological detection and imaging evaluation, a clinical prediction model is constructed based on a large cohort to improve the accuracy of distinguishing between benign and malignant pulmonary nodules. Methods A retrospective analysis was conducted on the clinical data of the 32 627 patients with pulmonary nodules who underwent chest CT and testing for 7 types of lung cancer-related serum autoantibodies (7-AABs) at our hospital from January 2020 to April 2024. The univariate and multivariate logistic regression models were performed to screen independent risk factors for benign and malignant pulmonary nodules, based on which a nomogram model was established. The performance of the model was evaluated using receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA). Results A total of 1 017 patients with pulmonary nodules were included in the study. The training set consisted of 712 patients, including 291 males and 421 females, with a mean age of (58±12) years. The validation set included 305 patients, comprising 129 males and 176 females, with a mean age of (58±13) years. Univariate ROC curve analysis indicated that the combination of CT and 7-AABs testing achieved the highest area under the curve (AUC) value (0.794), surpassing the diagnostic efficacy of CT alone (AUC=0.667) or 7-AABs alone (AUC=0.514). Multivariate logistic regression analysis showed that radiological nodule diameter, nodule nature, and CT combined with 7-AABs detection were independent predictors, which were used to construct a nomogram prediction model. The AUC values for this model were 0.826 and 0.862 in the training and validation sets, respectively, demonstrating excellent performance in DCA. Conclusion The combination of 7-AABs with CT significantly enhances the accuracy of distinguishing between benign and malignant pulmonary nodules. The developed predictive model provides strong support for clinical decision-making and contributes to achieving precise diagnosis and treatment of pulmonary nodules.
4.Synthetic MRI Combined With Clinicopathological Characteristics for Pretreatment Prediction of Chemoradiotherapy Response in Advanced Nasopharyngeal Carcinoma
Siyu CHEN ; Jiankun DAI ; Jing ZHAO ; Shuang HAN ; Xiaojun ZHANG ; Jun CHANG ; Donghui JIANG ; Heng ZHANG ; Peng WANG ; Shudong HU
Korean Journal of Radiology 2025;26(2):135-145
Objective:
To explore the feasibility of synthetic magnetic resonance imaging (syMRI) combined with clinicopathological characteristics for the pre-treatment prediction of chemoradiotherapy (CRT) response in advanced nasopharyngeal carcinoma (ANPC).
Materials and Methods:
Patients with ANPC treated with CRT between September 2020 and June 2022 were retrospectively enrolled and categorized into response group (RG, n = 95) and non RGs (NRG, n = 32) based on the Response Evaluation Criteria in Solid Tumors (RECIST) 1.1. The quantitative parameters from pre-treatment syMRI (longitudinal [T1] and transverse [T2] relaxation times and proton density [PD]), diffusion-weighted imaging (apparent diffusion coefficient [ADC]), and clinicopathological characteristics were compared between RG and NRG. Logistic regression analysis was applied to identify parameters independently associated with CRT response and to construct a multivariable model. The areas under the receiveroperating characteristic curve (AUC) for various diagnostic approaches were compared using the DeLong test.
Results:
The T1, T2, and PD values in the NRG were significantly lower than those in the RG (all P < 0.05), whereas no significant difference was observed in the ADC values between these two groups. Clinicopathological characteristics (Epstein–Barr virus [EBV]-DNA level, lymph node extranodal extension, clinical stage, and Ki-67 expression) exhibited significant differences between the two groups. Logistic regression analysis showed that T1, PD, EBV-DNA level, clinical stage, and Ki-67 expression had significant independent relationships with CRT response (all P < 0.05). The multivariable model incorporating these five variables yielded AUC, sensitivity, and specificity values of 0.974, 93.8% (30/32), and 91.6% (87/95), respectively.
Conclusion
SyMRI may be used for the pretreatment prediction of CRT response in ANPC. The multivariable model incorporating syMRI quantitative parameters and clinicopathological characteristics, which were independently associated with CRT response, may be a new tool for the pretreatment prediction of CRT response.
5.Shenqi Dihuang Decoction Improves Renal Function in Mouse Model of Diabetic Kidney Disease by Inhibiting Arachidonic Acid-related Ferroptosis Via ACSL4/LPCAT3/ALOX15 Axis
Yuantao WU ; Zhibin WANG ; Xinying FU ; Xiaoling ZOU ; Wenxiao HU ; Yixian ZOU ; Jun FENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):140-149
ObjectiveTo investigate the therapeutic effects and mechanism of Shenqi Dihuang decoction (SQDHD) on diabetic kidney disease (DKD), with a focus on its impact on arachidonic acid-related ferroptosis. MethodsSixty C57BL/6 mice were allocated into a normal group (n=10) and a modeling group (n=50), with 43 mice successfully modeled. The successfully modeled mice were further allocated into model, low-, medium-, and high-dose (4.68, 9.36, and 18.72 g·kg-1, respectively) SQDHD, and dapagliflozin (0.13 mg·kg-1) groups. The drug treatment groups were administrated with corresponding agents by gavage, and the normal and model groups were administrated with equal volumes of normal saline by gavage. An electronic balance and a glucometer were used to monitor the body weight and fasting blood glucose level from the tail tip, respectively. Serum creatinine (Scr) and blood urea nitrogen (BUN) levels were measured by enzyme-linked immunosorbent assay (ELISA). Histopathological changes in the renal tissue were assessed by hematoxylin-eosin staining, Masson staining, and periodic acid-Schiff (PAS) staining. The fluorescence intensity of reactive oxygen species (ROS) in frozen sections was observed by an inverted fluorescence microscope to evaluate the levels of ferrous ions (Fe2+) and lipid peroxidation in the renal tissue. Immunofluorescence staining of glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4) in the renal tissue was performed to detect their localization and expression. Western blot was employed to assess the expression levels of key ferroptosis proteins such as GPX4 and cystine/glutamate antiporter (xCT), as well as the arachidonic acid metabolic pathway-related proteins, including ACSL4, lysophosphatidylcholine acyltransferase 3 (LPCAT3), and arachidonate 15-lipoxygenase (ALOX15). Real-time PCR was employed to measure the mRNA levels of key ferroptosis proteins, including solute carrier family 7 member 11 (SLC7A11) and GPX4, as well as arachidonic acid metabolism-related factors (ACSL4, LPCAT3, and ALOX15) in the renal tissue. ResultsCompared with the normal group, DKD model mice exhibited a decrease in body weight (P<0.01), increases in levels of blood glucose (P<0.01), 24-hour urinary protein, Scr, and BUN (P<0.01), along with severe pathological changes, such as mesangial cell proliferation, basement membrane thickening, tubular atrophy, and interstitial inflammatory cell infiltration. In addition, the modeling elevated the levels of Fe2+, MDA, LPO, and ROS (P<0.01), lowered the GPX4 and xCT levels (P<0.01), raised the ACSL4, LPCAT3, and ALOX15 levels (P<0.01), down-regulated the mRNA levels of GPX4 and SLC7A11 (P<0.01), and up-regulated the mRNA levels of ACSL4, LPCAT3, and ALOX15 (P<0.01) in the renal tissue. Compared with the model group, low-, medium-, and high-dose SQDHD groups and the dapagliflozin group showed an increase in body weight (P<0.01), decreases in levels of blood glucose (P<0.01), 24-hour urinary protein, and Scr (P<0.01), alleviated pathological changes in glomeruli and tubules, and reduced degree of glomerular and tubular fibrosis. The high-dose SQDHD group and the dapagliflozin group showed reductions in Fe2+, MDA, LPO, and ROS levels (P<0.01). The medium- and high-dose SQDHD groups and the dapagliflozin group exhibited increased levels of GPX4 and xCT (P<0.01), decreased levels of ACSL4, LPCAT3, and ALOX15 (P<0.05, P<0.01), and down-regulated mRNA levels of ACSL4, LPCAT3, and ALOX15 (P<0.01). ConclusionSQDHD ameliorates DKD by inhibiting ferroptosis potentially by reducing iron ion levels, inhibiting lipid peroxidation, up-regulating GPX4 expression, and down-regulating ACSL4 expression. This study provides new insights and a theoretical basis for the treatment of DKD with traditional Chinese medicine and identifies potential targets for developing novel therapeutics for DKD.
6.Shenqi Dihuang Decoction Improves Renal Function in Mouse Model of Diabetic Kidney Disease by Inhibiting Arachidonic Acid-related Ferroptosis Via ACSL4/LPCAT3/ALOX15 Axis
Yuantao WU ; Zhibin WANG ; Xinying FU ; Xiaoling ZOU ; Wenxiao HU ; Yixian ZOU ; Jun FENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):140-149
ObjectiveTo investigate the therapeutic effects and mechanism of Shenqi Dihuang decoction (SQDHD) on diabetic kidney disease (DKD), with a focus on its impact on arachidonic acid-related ferroptosis. MethodsSixty C57BL/6 mice were allocated into a normal group (n=10) and a modeling group (n=50), with 43 mice successfully modeled. The successfully modeled mice were further allocated into model, low-, medium-, and high-dose (4.68, 9.36, and 18.72 g·kg-1, respectively) SQDHD, and dapagliflozin (0.13 mg·kg-1) groups. The drug treatment groups were administrated with corresponding agents by gavage, and the normal and model groups were administrated with equal volumes of normal saline by gavage. An electronic balance and a glucometer were used to monitor the body weight and fasting blood glucose level from the tail tip, respectively. Serum creatinine (Scr) and blood urea nitrogen (BUN) levels were measured by enzyme-linked immunosorbent assay (ELISA). Histopathological changes in the renal tissue were assessed by hematoxylin-eosin staining, Masson staining, and periodic acid-Schiff (PAS) staining. The fluorescence intensity of reactive oxygen species (ROS) in frozen sections was observed by an inverted fluorescence microscope to evaluate the levels of ferrous ions (Fe2+) and lipid peroxidation in the renal tissue. Immunofluorescence staining of glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4) in the renal tissue was performed to detect their localization and expression. Western blot was employed to assess the expression levels of key ferroptosis proteins such as GPX4 and cystine/glutamate antiporter (xCT), as well as the arachidonic acid metabolic pathway-related proteins, including ACSL4, lysophosphatidylcholine acyltransferase 3 (LPCAT3), and arachidonate 15-lipoxygenase (ALOX15). Real-time PCR was employed to measure the mRNA levels of key ferroptosis proteins, including solute carrier family 7 member 11 (SLC7A11) and GPX4, as well as arachidonic acid metabolism-related factors (ACSL4, LPCAT3, and ALOX15) in the renal tissue. ResultsCompared with the normal group, DKD model mice exhibited a decrease in body weight (P<0.01), increases in levels of blood glucose (P<0.01), 24-hour urinary protein, Scr, and BUN (P<0.01), along with severe pathological changes, such as mesangial cell proliferation, basement membrane thickening, tubular atrophy, and interstitial inflammatory cell infiltration. In addition, the modeling elevated the levels of Fe2+, MDA, LPO, and ROS (P<0.01), lowered the GPX4 and xCT levels (P<0.01), raised the ACSL4, LPCAT3, and ALOX15 levels (P<0.01), down-regulated the mRNA levels of GPX4 and SLC7A11 (P<0.01), and up-regulated the mRNA levels of ACSL4, LPCAT3, and ALOX15 (P<0.01) in the renal tissue. Compared with the model group, low-, medium-, and high-dose SQDHD groups and the dapagliflozin group showed an increase in body weight (P<0.01), decreases in levels of blood glucose (P<0.01), 24-hour urinary protein, and Scr (P<0.01), alleviated pathological changes in glomeruli and tubules, and reduced degree of glomerular and tubular fibrosis. The high-dose SQDHD group and the dapagliflozin group showed reductions in Fe2+, MDA, LPO, and ROS levels (P<0.01). The medium- and high-dose SQDHD groups and the dapagliflozin group exhibited increased levels of GPX4 and xCT (P<0.01), decreased levels of ACSL4, LPCAT3, and ALOX15 (P<0.05, P<0.01), and down-regulated mRNA levels of ACSL4, LPCAT3, and ALOX15 (P<0.01). ConclusionSQDHD ameliorates DKD by inhibiting ferroptosis potentially by reducing iron ion levels, inhibiting lipid peroxidation, up-regulating GPX4 expression, and down-regulating ACSL4 expression. This study provides new insights and a theoretical basis for the treatment of DKD with traditional Chinese medicine and identifies potential targets for developing novel therapeutics for DKD.
7.Application of blood conservation measures with different red blood cell transfusion volumes in obstetrics and their impact on postpartum outcomes
Huimin DENG ; Fengcheng XU ; Meiting LI ; Lan HU ; Xiao WANG ; Shiyu WANG ; Xiaofei YUAN ; Jun ZHENG ; Zehua DONG ; Yuanshan LU ; Shaoheng CHEN
Chinese Journal of Blood Transfusion 2025;38(5):691-698
Objective: To evaluate the application of blood conservation measures in obstetric patients with different red blood cell transfusion volumes and to assess the impact of different transfusion volumes on postpartum outcomes. Methods: A retrospective investigation was conducted on 448 obstetric patients who received blood transfusions at the Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine from January 2016 to December 2022. Patients were divided into four groups (1-2 units group, 3-4 units group, 5-6 units group, and >6 units group) based on the volumes of red blood cells (RBCs) transfused during and within 7 days after delivery. The maternal physiological indicators, pre- and postpartum laboratory test indicators, obstetric complications, application of blood conservation measures, use of blood products, and postpartum outcomes were reviewed. The clinical characteristics, application of blood conservation measures, and their impact on postpartum outcomes were compared among different transfusion groups. Results: There were statistically significant differences in the multivariate logistic analysis of history of previous cesarean section (OR=1.781), eclampsia/pre-eclampsia/(OR=1.972) and postpartum blood loss>1 000 mL(OR=1.699)(P<0.05) among different transfusion groups. In terms of blood conservation measures, the more RBCs transfused, the higher the rate of mothers receiving blood conservation measures such as balloon occlusion, arterial ligation, autologous blood transfusion with a cell saver, and hysterectomy. With the increase in the volume of RBCs transfusion, the demand for fresh frozen plasma(FFP), cryoprecipitate, and platelet transfusions also increased. The hospitalization days for the four groups of parturients were 6.0 (4.0-9.0), 7.5 (5.0-14.8), 7.0 (4.5-13.0) and 11.0 (9.0-20.5), respectively (P<0.05) and the rates of ICU transfer were 2.0% (5/250), 9.4% (12/128),18.2% (6/33) and 51.4% (19/37), respectively (P<0.05). Both increased significantly with the increase in the volume of RBCs transfusion, and the differences between groups were statistically significant. Conclusion: Parturients who received higher volume of RBCs had multiple risks factors for bleeding before childbirth, had higher postpartum blood loss, and had a higher rate of application of various blood conservation measures. In addition, an increase in the volume of RBCs transfusion may have adverse effects on postpartum recovery.
8.Integrated molecular characterization of sarcomatoid hepatocellular carcinoma
Rong-Qi SUN ; Yu-Hang YE ; Ye XU ; Bo WANG ; Si-Yuan PAN ; Ning LI ; Long CHEN ; Jing-Yue PAN ; Zhi-Qiang HU ; Jia FAN ; Zheng-Jun ZHOU ; Jian ZHOU ; Cheng-Li SONG ; Shao-Lai ZHOU
Clinical and Molecular Hepatology 2025;31(2):426-444
Background:
s/Aims: Sarcomatoid hepatocellular carcinoma (HCC) is a rare histological subtype of HCC characterized by extremely poor prognosis; however, its molecular characterization has not been elucidated.
Methods:
In this study, we conducted an integrated multiomics study of whole-exome sequencing, RNA-seq, spatial transcriptome, and immunohistochemical analyses of 28 paired sarcomatoid tumor components and conventional HCC components from 10 patients with sarcomatoid HCC, in order to identify frequently altered genes, infer the tumor subclonal architectures, track the genomic evolution, and delineate the transcriptional characteristics of sarcomatoid HCCs.
Results:
Our results showed that the sarcomatoid HCCs had poor prognosis. The sarcomatoid tumor components and the conventional HCC components were derived from common ancestors, mostly accessing similar mutational processes. Clonal phylogenies demonstrated branched tumor evolution during sarcomatoid HCC development and progression. TP53 mutation commonly occurred at tumor initiation, whereas ARID2 mutation often occurred later. Transcriptome analyses revealed the epithelial–mesenchymal transition (EMT) and hypoxic phenotype in sarcomatoid tumor components, which were confirmed by immunohistochemical staining. Moreover, we identified ARID2 mutations in 70% (7/10) of patients with sarcomatoid HCC but only 1–5% of patients with non-sarcomatoid HCC. Biofunctional investigations revealed that inactivating mutation of ARID2 contributes to HCC growth and metastasis and induces EMT in a hypoxic microenvironment.
Conclusions
We offer a comprehensive description of the molecular basis for sarcomatoid HCC, and identify genomic alteration (ARID2 mutation) together with the tumor microenvironment (hypoxic microenvironment), that may contribute to the formation of the sarcomatoid tumor component through EMT, leading to sarcomatoid HCC development and progression.
9.Guidelines for vaccination of kidney transplant candidates and recipients in China
Jian Zhang ; Jun Lin ; Weijie Zhang ; Xiaoming Ding ; Xiaopeng Hu ; Wujun Xue
Organ Transplantation 2025;16(2):177-190
In order to further standardize the vaccination of kidney transplant candidates and recipients in China, the Branch of Organ Transplantation of Chinese Medical Association has organized experts in kidney transplantation and infectious diseases. Based on the "Vaccination of Solid Organ Transplant Candidates and Recipients: Guidelines from the American Society of Transplantation Infectious Diseases Community of Practice", and in combination with the clinical reality of infectious diseases and vaccination after organ transplantation in China, as well as referring to relevant recommendations from home and abroad in recent years, these guidelines are formulated from aspects such as epidemiology, types of vaccines, vaccination principles, target population, and specific vaccine administration. The "Guidelines for Vaccination of Kidney Transplant Candidates and Recipients in China" aims to provide theoretical reference for medical workers in the field of kidney transplantation in China, regarding the vaccination of kidney transplant candidates and recipients. It is expected to better guide the vaccination of kidney transplant candidates and recipients, reduce the risk of postoperative infection, and improve survival outcomes.
10.Effect of Hei Xiaoyaosan on Neuroinflammation and NLRP3/Caspase-1/GSDMD Signaling Pathway in APP/PS1 Mice
Jun ZHOU ; Mingcheng LI ; Yujie LYU ; Zhipeng MENG ; Yunyun HU ; Huping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):124-133
ObjectiveTo observe the effects of Hei Xiaoyaosan on the learning and memory abilities of Alzheimer's disease model mice (APP/PS1 mice), and to explore its mechanism through the inflammatory cascade mediated by nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3)/cysteine aspartate-specific protease (Caspase-1)/gasdermin D (GSDMD) signaling pathway. MethodsSPF-grade 4-month-old APP/PS1 mice were randomly divided into the model group, MCC950 group, and Hei Xiaoyaosan high-, medium-, and low-dose groups. C57BL/6J mice were used as the blank group. After 7 days of adaptive feeding, mice in each group were intervened. The Hei Xiaoyaosan high-, medium-, and low-dose groups were given corresponding doses by gavage (25.79, 12.90, 6.45 g·kg-1·d-1), the MCC950 group was intraperitoneally injected with 10 mg·kg-1·2 d-1, and the blank group received the same volume of physiological saline by gavage. After 90 days of intervention, the learning and memory abilities were assessed using the Y maze and Morris water maze tests. The structural changes of hippocampal neurons were observed by hematoxylin-eosin (HE) staining. The expression of amyloid precursor protein (APP) in the hippocampal CA3 region was detected by immunohistochemistry. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interleukin (IL)-10, IL-18, and IL-1β in the hippocampus. Western blot was applied to detect the protein expression of NLRP3, Caspase-1, GSDMD, and GSDMD-N in the hippocampus. Immunofluorescence was used to detect the co-localization of GSDMD-N and ionized calcium-binding adapter molecule-1 (Iba-1) in the hippocampus. Results① In the Y maze test, compared with the blank group, the spontaneous alternation rate of the model group was significantly reduced (P<0.01). Compared with the model group, the spontaneous alternation rate in the Hei Xiaoyaosan high- and low-dose groups was significantly increased (P<0.01). ② In the Morris water maze test, during the 1-4 days of the location navigation test, the escape latency time of mice decreased with the extension of training time. On day 4, compared with the blank group, the model group showed a significantly increased escape latency (P<0.05). Compared with the model group, the MCC950 group and the Hei Xiaoyaosan low-dose group showed significantly reduced escape latency (P<0.05). In the spatial exploration experiment, compared with the blank group, the number of platform crossings in the model group was significantly reduced (P<0.01). Compared with the model group, the Hei Xiaoyaosan low-dose group showed significantly increased platform crossings (P<0.05). ③ HE staining showed that, compared with the blank group, the hippocampal CA3 cells of the model group were damaged, arranged loosely and irregularly, swollen, with unclear boundaries, and the nuclei were pyknotic and deeply stained. MCC950 and all doses of Hei Xiaoyaosan improved the hippocampal CA3 cell damage in APP/PS1 mice to varying degrees. ④ Immunohistochemical results indicated that, compared with the blank group, the expression of APP in the hippocampal CA3 region was significantly increased in the model group (P<0.01). MCC950 and all doses of Hei Xiaoyaosan could reduce the expression of APP in the hippocampal CA3 region of APP/PS1 mice (P<0.01). ⑤ ELISA results showed that the levels of IL-18 and IL-1β in the hippocampus of mice in the model group were significantly increased, and IL-10 levels were significantly reduced (P<0.01). Compared with the model group, the IL-18 levels in the MCC950 group and the Hei Xiaoyaosan medium- and low-dose groups were significantly reduced (P<0.01). IL-1β levels in the hippocampus of the MCC950 group and Hei Xiaoyaosan high-, medium-, and low-dose groups were significantly decreased (P<0.01). The IL-10 levels in the hippocampus of the MCC950 group and the Hei Xiaoyaosan medium- and low-dose groups were increased (P<0.05, P<0.01). ⑥ Western blot results showed that compared with the blank group, the protein levels of NLRP3, Caspase-1, GSDMD, and GSDMD-N in the hippocampus of the model group were significantly elevated (P<0.01). Compared with the model group, the content of NLRP3 and Caspase-1 in the hippocampus of the treated groups was decreased (P<0.05, P<0.01). The content of GSDMD in the hippocampus of the Hei Xiaoyaosan high-, medium-, and low-dose groups was reduced (P<0.05, P<0.01), and the content of GSDMD-N in the hippocampus of the Hei Xiaoyaosan medium- and low-dose groups was decreased (P<0.05, P<0.01). ⑦ Immunofluorescence results showed that, compared with the blank group, the co-expression of GSDMD-N and Iba-1 in the hippocampus of the model group was significantly increased (P<0.01). Compared with the model group, the co-expression of GSDMD-N and Iba-1 in the treated groups was significantly reduced (P<0.01). ConclusionHei Xiaoyaosan may regulate the NLRP3/Caspase-1/GSDMD signaling pathway to affect the release of inflammatory factors, alleviate neuroinflammation,improve hippocampal histopathological changes,and improve learning and memory deficits,thus providing potential therapeutic benefits for Alzheimer's disease.

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