1.Sesquiterpene ZH-13 from Aquilariae Lignum Resinatum Improves Neuroinflammation by Regulating JNK Phosphorylation
Ziyu YIN ; Yun GAO ; Junjiao WANG ; Weigang XUE ; Xueping PANG ; Huiting LIU ; Yunfang ZHAO ; Huixia HUO ; Jun LI ; Jiao ZHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):139-145
ObjectiveTo study the pharmacological substances and mechanisms through which sesquiterpene ZH-13 from Aquilariae Lignum Resinatum improves neuroinflammation. MethodsBV-2 microglial cells were stimulated with lipopolysaccharide (LPS) to induce neuroinflammation. The cells were divided into the normal group, the model group, and the ZH-13 low- and high-dose treatment groups (10, 20 μmol·L-1). The model group was treated with 1 μmol·L-1 LPS. Cell viability was assessed using the cell proliferation and activity assay (CCK-8 kit). Nitric oxide (NO) release in the cell supernatant was measured using a nitric oxide kit (Griess method). The mRNA expression levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and interleukin-6 (IL-6) were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The phosphorylation of mitogen-activated protein kinase (MAPK) pathway proteins was assessed by Western blot. ResultsCompared with the model group, ZH-13 dose-dependently reduced NO release from BV-2 cells under LPS stimulation (P<0.05, P<0.01). In the 20 μmol·L-1 ZH-13 treatment group, the mRNA expression levels of IL-1β, TNF-α, iNOS, and IL-6 were significantly reduced compared to the model group (P<0.05, P<0.01). In both the low- and high-dose ZH-13 groups, the expression of the inflammatory factor TNF-α and the phosphorylation of c-Jun N-terminal kinase (JNK) in the upstream MAPK pathway were significantly reduced (P<0.05). After stimulation with the JNK agonist anisomycin (Ani), both low- and high-dose ZH-13 treatment groups showed reduced phosphorylation of JNK proteins compared to the Ani-treated group (P<0.01). ConclusionThe sesquiterpene compound ZH-13 from Aquilariae Lignum Resinatum significantly ameliorates LPS-induced neuroinflammatory responses in BV-2 cells by inhibiting excessive JNK phosphorylation and reducing TNF-α expression. These findings elucidate the pharmacological substances and mechanisms underlying the sedative and calming effects of Aquilariae Lignum Resinatum.
2.Effect of Wenyang Shengji Ointment (温阳生肌膏) on MGO Content and HIF-1a/VEGF Pathway in Wound Tissue of Diabetic Wound Model Rats with Yin Syndrome
Xinyu HUANG ; Li CHEN ; Yarong DING ; Jun WANG ; Shuihua FENG ; Zhongzhi ZHOU
Journal of Traditional Chinese Medicine 2025;66(4):382-389
ObjectiveTo investigate the possible mechanism of Wenyang Shengji Ointment (温阳生肌膏, WSO) in the treatment of diabetic wounds with yin syndrome. MethodsA total of 24 SD rats were randomly divided into a group (n=6) and modeling group (n=18). The modeling group rats were fed with high-fat diet for 14 days and then were injected intraperitoneally with streptozotocin to induce diabetic model. After steroid injection, full-thickness skin defects were created on the back of the rats to establish a diabetic wound with yin syndrome model. The normal group was fed with regular diet, and full-thickness skin defects were created surgically on the back of the rats. The 18 successfully modeled rats were further divided into three groups, the model group, the WSO group, and the Beifuxin (Recombinant Bovine Basic Fibroblast Growth Factor Gel, BX) group, 6 rats in each group. The WSO group was given the ointment to the wound, the Beifuxin group was givne BX gel, and the normal group and model group was disinfected and treated with saline. All groups had their dressings changed once daily for 14 days. Wound healing was recorded on days 0, 3, 7, and 14, and the wound healing rate was calculated on day 3, 7, and 14. On day 14 after treatment, HE staining was performed to observe the pathological morphology of the wound tissue. Western Blot was used to detect the relative protein levels of hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF). Immunofluorescence was used to measure the fluorescence intensity of HIF-1α in the wound tissue, and ELISA was used to detect the methylglyoxal (MGO) content in the wound tissue. ResultsCompared with the normal group, the model group showed poor wound healing on day 3, 7, and 14, with a low wound healing rate (P<0.01). HE staining showed scab coverage on the wound, with inflammatory cell infiltration and disorganized collagen arrangement. The relative protein levels of VEGF were significantly reduced, while the relative protein levels of HIF-1α and the MGO content significantly increased (P<0.01), and the fluorescence intensity of HIF-1α was enhanced. Compared to the model group, the WSO group and Beifuxin group showed better wound healing on day 3, 7, and 14, with an increased wound healing rate (P<0.01). The wound tissue showed clear and complete epithelial structure, reduced inflammatory cells, mature granulation tissue, and organized collagen arrangement. MGO content was significantly reduced (P<0.01). The relative protein levels of HIF-1α and VEGF both significantly increased in the WSO group, while only VEGF increased in the Beifuxin group (P<0.05 or P<0.01). Compared with the Beifuxin group, the WSO group had a thicker epidermal layer, prominent collagen formation, significantly increased HIF-1α fluorescence expression, reduced MGO content in the wound tissue, and higher relative protein levels of HIF-1α (P<0.05). ConclusionWSO can reduce the accumulation of MGO in diabetic wound tissue with yin syndrome and activate the HIF-1α/VEGF pathway, which could be one of the mechanisms for promoting wound healing.
3.Exploring Vascular Recruitment in Tracheobronchial Adenoid Cystic Carcinoma from Perspective of Abnormal Collateral
Jun TENG ; Lei LI ; Junyan XIA ; Yi LUO ; Qinyan HONG ; Shuiping CHEN ; Hongwu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):260-269
Tracheobronchial adenoid cystic carcinoma (TACC) is a low-grade malignant tumor originating from the airway mucosa. Despite its slow progression,it is characterized by high invasiveness,frequent recurrence,and a strong tendency for metastasis. Preclinical studies have shown that vascular-targeted therapy holds significant potential. However,an effective systemic treatment for TACC has not been established yet. This study explored TACC from the perspective of "Feiji" in traditional Chinese medicine (TCM) as the starting point. It deeply investigated the mechanisms of abnormal collaterals and tumor vascular recruitment and further elaborated on the theoretical connection between abnormal collaterals and tumor vascular recruitment. Firstly,collateral hyperactivity led to disordered and erratic pulmonary collaterals. Their abnormal structures were similar to the disorderly and tortuous nature of tumor (pseudo)angiogenesis. This resulted in imbalances in the functions of circulation,perfusion,and reverse injection of the pulmonary collaterals,and then led to unrestrained collateral dysfunction and the accumulation of pathogenic factors. Secondly,the remodeling of the extracellular matrix (ECM) and epithelial-mesenchymal transition (EMT) in TACC were critical processes in vascular co-option (VCO),representing the micro-level manifestation of the displacement of nutrient and defense. During this process,ECM remodeling made TACC cells more likely to hijack normal blood vessels,creating a complex vascular microenvironment conducive to tumor growth. In terms of treatment,this study proposed a TCM strategy of "regulating collaterals to expel pathogenic factors and nourishing collaterals to strengthen the healthy Qi",and listed potential TCM. These were intended to regulate the Qi and blood in the collaterals,repair the functions of abnormal collaterals,and intervene in the vascular recruitment process of TACC. Future research should focus on improving the TCM clinical syndrome characteristics of TACC. Through modern molecular biology techniques,it is necessary to deeply analyze the micro-level pattern of vascular recruitment in TACC. This would enrich the understanding of the profound connection between abnormal collaterals and tumor vascular recruitment,providing empirical evidence for TCM-targeted therapies for vascular recruitment in TACC.
4.Translational Research of Electromagnetic Fields on Diseases Related With Bone Remodeling: Review and Prospects
Peng SHANG ; Jun-Yu LIU ; Sheng-Hang WANG ; Jian-Cheng YANG ; Zhe-Yuan ZHANG ; An-Lin LI ; Hao ZHANG ; Yu-Hong ZENG
Progress in Biochemistry and Biophysics 2025;52(2):439-455
Electromagnetic fields can regulate the fundamental biological processes involved in bone remodeling. As a non-invasive physical therapy, electromagnetic fields with specific parameters have demonstrated therapeutic effects on bone remodeling diseases, such as fractures and osteoporosis. Electromagnetic fields can be generated by the movement of charged particles or induced by varying currents. Based on whether the strength and direction of the electric field change over time, electromagnetic fields can be classified into static and time-varying fields. The treatment of bone remodeling diseases with static magnetic fields primarily focuses on fractures, often using magnetic splints to immobilize the fracture site while studying the effects of static magnetic fields on bone healing. However, there has been relatively little research on the prevention and treatment of osteoporosis using static magnetic fields. Pulsed electromagnetic fields, a type of time-varying field, have been widely used in clinical studies for treating fractures, osteoporosis, and non-union. However, current clinical applications are limited to low-frequency, and research on the relationship between frequency and biological effects remains insufficient. We believe that different types of electromagnetic fields acting on bone can induce various “secondary physical quantities”, such as magnetism, force, electricity, acoustics, and thermal energy, which can stimulate bone cells either individually or simultaneously. Bone cells possess specific electromagnetic properties, and in a static magnetic field, the presence of a magnetic field gradient can exert a certain magnetism on the bone tissue, leading to observable effects. In a time-varying magnetic field, the charged particles within the bone experience varying Lorentz forces, causing vibrations and generating acoustic effects. Additionally, as the frequency of the time-varying field increases, induced currents or potentials can be generated within the bone, leading to electrical effects. When the frequency and power exceed a certain threshold, electromagnetic energy can be converted into thermal energy, producing thermal effects. In summary, external electromagnetic fields with different characteristics can generate multiple physical quantities within biological tissues, such as magnetic, electric, mechanical, acoustic, and thermal effects. These physical quantities may also interact and couple with each other, stimulating the biological tissues in a combined or composite manner, thereby producing biological effects. This understanding is key to elucidating the electromagnetic mechanisms of how electromagnetic fields influence biological tissues. In the study of electromagnetic fields for bone remodeling diseases, attention should be paid to the biological effects of bone remodeling under different electromagnetic wave characteristics. This includes exploring innovative electromagnetic source technologies applicable to bone remodeling, identifying safe and effective electromagnetic field parameters, and combining basic research with technological invention to develop scientifically grounded, advanced key technologies for innovative electromagnetic treatment devices targeting bone remodeling diseases. In conclusion, electromagnetic fields and multiple physical factors have the potential to prevent and treat bone remodeling diseases, and have significant application prospects.
5.Study on quality evaluation of Mongolian medicine Sanzi powder:fingerprint,chemical pattern recognition and multi-component quantification analysis
Jun LI ; Rongjie LI ; Fengye ZHOU ; Qian ZHANG ; Wei ZHANG ; Bohan ZHANG ; Shu WANG ; Xitong ZHAO ; Jianping CHEN
China Pharmacy 2025;36(4):414-420
OBJECTIVE To establish fingerprint, chemical pattern recognition and multi-component quantification analysis of Sanzi powder, and evaluate its quality. METHODS HPLC method was adopted. The fingerprints of 15 batches of Sanzi powder were established by using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 edition). Cluster analysis, principal component analysis and orthogonal partial least squares-discriminant analysis were also conducted. The variable importance in projection (VIP) value greater than 1 was used as the index to screen the differential markers, and the contents of the differential markers were determined by the same HPLC method. RESULTS A total of 21 common peaks in the HPLC fingerprints of 15 batches of Sanzi powder were calibrated, and the similarities of them were 0.994- 0.999; 6 common peaks were identified, including gallic acid (peak 3), garminoside (peak 10), corilagin (peak 11), chebulinic acid (peak 16), ellagic acid (peak 18), crocin Ⅰ (peak 19). According to the results of cluster analysis, YKD2024LH005,No.YKD2023LH062) principal component analysis and orthogonal partial least squares-discriminant analysis, 15 batches of samples could be clustered into two categories: S1, S5, S7, S9, S14 were clustered into one category; S2-S4, S6, S8, S10-S13, S15 were clustered into one category. VIP values of 11 differential components such as corilagin, chebulinic acid and ellagic acid were higher than 1. Among 15 batches of samples, the contents of corilagin, chebulinic acid and ellagic acid ranged 2.667-5.152, 9.506- 13.522, 0.891-1.811 mg/g. CONCLUSIONS Established HPLC fingerprint and multi-component quantification analysis of Sanzi powder are rapid and simple, and can be used for quality evaluation of Sanzi powder by combining with chemical pattern recognition. Eleven components such as corilagin, chebulinic acid and ellagic acid are differential markers affecting the quality of Sanzi powder.
6.Connotation and Prevention Strategies of Traditional Chinese Medicine for Panvascular Diseases
Jie WANG ; Jun LI ; Yan DONG ; Cong CHEN ; Yongmei LIU ; Chao LIU ; Lanchun LIU ; Xuan SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):1-14
Panvascular disease, with vascular diseases as the common pathological feature, is mainly manifested as atherosclerosis. Panvascular disease mainly affects the important organs of the heart, brain, kidney, and limbs. It is one of the leading causes of death for Chinese residents at present. Previously, due to the narrow branches of disciplines, too much attention was paid to local lesions, resulting in the neglect of panvascular disease as a systemic one. The fact that panvascular disease has overall pathology and comprehensive and individualized treatment strategies, makes the disease highly compatible with the principles of holism concept and syndrome differentiation and treatment in traditional Chinese medicine (TCM). It is believed that blood stasis is the core pathogenesis of atherosclerosis and is involved in the whole process of atherosclerosis. The theories of ''blood vessel'', ''meridians'', ''visceral manifestation'', and ''organs-meridians'' in TCM are helpful to comprehensively understand the complexity of panvascular diseases. Moreover, those theories can provide systematic treatment strategies. The TCM syndromes of panvascular diseases evolve from ''phlegm, stasis, stagnation, and deficiency''. Panvascular arteriosclerosis is related to the syndrome of ''stasis and phlegm'', and the treatment mainly promotes blood circulation and removes phlegm. There are different specific drugs and mechanisms of action for coronary atherosclerosis, cerebral atherosclerosis, and renal artery atherosclerotic stenosis. Panvascular venous lesions are related to the syndrome of ''deficiency and stasis'' in TCM, and the TCM treatment mainly invigorates Qi and promotes blood circulation, which can inhibit venous thrombosis, improve venous ulcers, and resist venous endothelial damage. Panvascular microcirculatory lesions are inseparable from the ''stagnation and stasis'' in TCM, and the treatment mainly promotes Qi and dredges collaterals, which has a good effect on coronary microvascular lesions, diabetic microvascular lesions, pulmonary microvascular lesions, and pancreatic microvascular lesions. Panvascular lymphatic lesions are related to the syndrome of ''water and stasis'' in TCM. The treatment method focuses on promoting blood circulation and water excretion, which can promote lymphangiogenesis and enhance lymphatic reflux. In addition, the combination of TCM and modern technology, especially the application of artificial intelligence, can improve the efficiency of early identification and personalized treatment, resulting in early screening and comprehensive management of panvascular diseases. Therefore, TCM will play a vital role in the prevention and treatment of panvascular diseases.
7.Connotation and Prevention Strategies of Traditional Chinese Medicine for Panvascular Diseases
Jie WANG ; Jun LI ; Yan DONG ; Cong CHEN ; Yongmei LIU ; Chao LIU ; Lanchun LIU ; Xuan SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):1-14
Panvascular disease, with vascular diseases as the common pathological feature, is mainly manifested as atherosclerosis. Panvascular disease mainly affects the important organs of the heart, brain, kidney, and limbs. It is one of the leading causes of death for Chinese residents at present. Previously, due to the narrow branches of disciplines, too much attention was paid to local lesions, resulting in the neglect of panvascular disease as a systemic one. The fact that panvascular disease has overall pathology and comprehensive and individualized treatment strategies, makes the disease highly compatible with the principles of holism concept and syndrome differentiation and treatment in traditional Chinese medicine (TCM). It is believed that blood stasis is the core pathogenesis of atherosclerosis and is involved in the whole process of atherosclerosis. The theories of ''blood vessel'', ''meridians'', ''visceral manifestation'', and ''organs-meridians'' in TCM are helpful to comprehensively understand the complexity of panvascular diseases. Moreover, those theories can provide systematic treatment strategies. The TCM syndromes of panvascular diseases evolve from ''phlegm, stasis, stagnation, and deficiency''. Panvascular arteriosclerosis is related to the syndrome of ''stasis and phlegm'', and the treatment mainly promotes blood circulation and removes phlegm. There are different specific drugs and mechanisms of action for coronary atherosclerosis, cerebral atherosclerosis, and renal artery atherosclerotic stenosis. Panvascular venous lesions are related to the syndrome of ''deficiency and stasis'' in TCM, and the TCM treatment mainly invigorates Qi and promotes blood circulation, which can inhibit venous thrombosis, improve venous ulcers, and resist venous endothelial damage. Panvascular microcirculatory lesions are inseparable from the ''stagnation and stasis'' in TCM, and the treatment mainly promotes Qi and dredges collaterals, which has a good effect on coronary microvascular lesions, diabetic microvascular lesions, pulmonary microvascular lesions, and pancreatic microvascular lesions. Panvascular lymphatic lesions are related to the syndrome of ''water and stasis'' in TCM. The treatment method focuses on promoting blood circulation and water excretion, which can promote lymphangiogenesis and enhance lymphatic reflux. In addition, the combination of TCM and modern technology, especially the application of artificial intelligence, can improve the efficiency of early identification and personalized treatment, resulting in early screening and comprehensive management of panvascular diseases. Therefore, TCM will play a vital role in the prevention and treatment of panvascular diseases.
8.Impact of Onset-to-Door Time on Endovascular Therapy for Basilar Artery Occlusion
Tianlong LIU ; Chunrong TAO ; Zhongjun CHEN ; Lihua XU ; Yuyou ZHU ; Rui LI ; Jun SUN ; Li WANG ; Chao ZHANG ; Jianlong SONG ; Xiaozhong JING ; Adnan I. QURESHI ; Mohamad ABDALKADER ; Thanh N. NGUYEN ; Raul G. NOGUEIRA ; Jeffrey L. SAVER ; Wei HU
Journal of Stroke 2025;27(1):140-143
9.Aldehyde Dehydrogenase 2 Gene Mutation May Reduce the Risk of Rupture of Intracranial Aneurysm in Chinese Han Population
Xiheng CHEN ; Siming GUI ; Dachao WEI ; Dingwei DENG ; Yudi TANG ; Jian LV ; Wei YOU ; Jia JIANG ; Jun LIN ; Huijian GE ; Peng LIU ; Yuhua JIANG ; Lixin MA ; Yunci WANG ; Ming LV ; Youxiang LI
Journal of Stroke 2025;27(2):237-249
Background:
and Purpose Ruptured intracranial aneurysms (RIA) are associated with a mortality rate of up to 40% in the Chinese population, highlighting the critical need for targeted treatment interventions for at-risk individuals. Although the impact of aldehyde dehydrogenase 2 (ALDH2) gene mutations on susceptibility to intracranial aneurysms (IA) is well documented, the potential connection between ALDH2 rs671 single-nucleotide polymorphism (SNP) and RIA remains unexplored. Given the increased prevalence of ALDH2 gene mutations among Chinese Han individuals, it is clinically relevant to investigate the link between ALDH2 rs671 SNP and IA rupture.
Methods:
A prospective study was conducted on 546 patients diagnosed with IA to investigate the association between ALDH2 rs671 SNP and the risk of IA rupture.
Results:
The ALDH2 rs671 SNP (ALDH2*2) was significantly more prevalent in patients with unruptured IA (UIA) than in those with RIA (32.56% vs. 18.58%, P=0.004). Multivariate logistic regression analysis revealed that people with the ALDH2 mutation (ALDH2*1/*2 and ALDH2*2/*2 gene type) had a significantly reduced odds ratio (OR=0.49; 95% confidence level [CI] 0.27–0.88; P=0.018) for RIAs. Age-specific subgroup analysis indicated that the ALDH2 mutation provided a stronger protective effect in individuals aged 60 years and above with IA compared to those under 60 years old (OR=0.38 vs. OR=0.52, both P<0.05).
Conclusion
The incidence of RIA was significantly higher in individuals with a normal ALDH2 gene (ALDH2*1/*1) than in those with an ALDH2 rs671 SNP (ALDH2*1/*2 or ALDH2*2/*2). ALDH2 rs671 SNP may serve as a protective factor against RIA in the Chinese Han population.
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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