BACKGROUND:Huanglian Wendan decoction (HLWD) is traditionally used to treat a syndrome characterized by symptoms such as insomnia, irritability, and digestive disturbances. Its historical application in addressing phlegm-heat-induced mental and emotional disorders offers both a cultural and clinical basis for investigating its potential mechanisms in alleviating depressive symptoms via gut-brain axis. PURPOSE:To explore the potential therapeutic mechanisms of HLWD in improving depression. METHODS:Behavioral outcomes were assessed using the SPT, TST, OFT, and SIT. Neurogenesis was evaluated by measuring dendritic length and neuronal intersections. Allopregnanolone levels were measured using ELISA. Inflammatory signaling pathways (TLR4, MyD88, NF-κB) and cytokines (IL-1β, TNF-α, IL-10, IL-4) in the hippocampus were analyzed via WB. The integrity of the intestinal barrier was further validated through immunofluorescence and histological examination. Finally, GW6471 was employed to investigate whether PPAR-α mediates the effects of HLWD. RESULTS:The UPLC analysis identified 42 constituents, primarily comprising flavonoids, alkaloids, and coumarins. Behavioral assays demonstrated that HLWD effectively ameliorated depressive-like behaviors in mice. Mechanistically, HLWD elevated the levels of DCX-positive neurogenesis markers, inhibited microglial activation, protected the intestinal mucosal barrier, and reduced inflammatory responses in both the colon and hippocampus. The RNA-seq results indicated an enrichment of the PPAR signaling pathway in colon. Consistent with this finding, HLWD significantly upregulated both the protein and mRNA levels of PPAR-α in the colon. Meanwhile, it increased the concentration of allopregnanolone in hippocampal via peripheral circulation. Notably, the administration of the PPAR-α antagonist GW6471 reversed these neuroprotective effects and the associated protein changes, thereby underscoring the critical role of PPAR-α in the therapeutic mechanism of HLWD. CONCLUSION:HLWD can elevate the levels of PPAR-α and allopregnanolone in the colon, subsequently influencing the levels of allopregnanolone in the hippocampus via peripheral circulation. This process may enhance neurological function and alleviate depressive symptoms.

BACKGROUND:Renal fibrosis is a central pathological process in chronic kidney disease (CKD). Although Haikun Shenxi Capsule (HKSX) has shown clinical efficacy in CKD, its precise molecular targets and mechanisms remain unclear. OBJECTIVE:This study aims to determine whether HKSX exerts antifibrotic effects in CKD by modulating NKD2 expression via METTL3-mediated m6A pathway. METHODS:A CKD mouse model was established by unilateral ureteral obstruction (UUO). Mice were divided into Sham, Model, low-dose HKSX, high-dose HKSX, and positive drug groups. Renal pathology and fibrosis were assessed by H&E, Masson's trichrome, and Sirius red staining. Renal function was evaluated by measuring serum creatinine (Scr) and blood urea nitrogen (BUN) levels. Inflammatory cytokines, oxidative stress markers, apoptosis-related proteins, fibrotic markers, NKD2, and METTL3 were detected by ELISA, qPCR, and Western blotting. Global m6A levels and NKD2 mRNA-specific m6A enrichment were measured by colorimetric assay and MeRIP-qPCR, respectively. Renal-specific METTL3-overexpressing mice were generated via lentivirus to validate target dependency. The role of the m6A reader IGF2BP3 was investigated using RIP-qPCR. RESULTS:HKSX significantly improved renal function and attenuated renal pathological injury, inflammation, oxidative stress, apoptosis, and fibrosis in UUO mice. Mechanistically, HKSX dose-dependently downregulated METTL3 expression, reduced both global renal m6A levels and NKD2 mRNA-specific m6A modification, and consequently inhibiting NKD2 overexpression. The protective effects of HKSX were largely abrogated in METTL3-overexpressing CKD mice. Furthermore, HKSX specifically inhibited the expression of the m6A reader IGF2BP3 and its binding to NKD2 mRNA. CONCLUSION:This study demonstrates that HKSX alleviates renal fibrosis by inhibiting METTL3, which reduces m6A modification on NKD2 mRNA, and by downregulating the stability reader IGF2BP3. These effects collectively decrease NKD2 mRNA stability and protein expression. These findings reveal a novel epitranscriptomic mechanism of HKSX and identify the METTL3/IGF2BP3/NKD2 axis as a key therapeutic pathway in CKD.

Sudden sensorineural hearing loss (SSNHL) is a common auditory disorder, with oxidative stress and ferroptosis implicated in its pathogenesis. Notoginsenoside R1, a major component of Panax notoginseng, exhibits antioxidant and cytoprotective effects, but its role in ferroptosis regulation remains unclear. In this study, a mouse model of SSNHL and an oxygen-glucose deprivation/reoxygenation (OGD/R)-induced Ear Institute-Organ of Corti 1 (HEI-OC1) cell model were used to evaluate the effects of Notoginsenoside R1. Notoginsenoside R1 significantly reduced auditory brainstem response (ABR) thresholds, reduced cochlear hair cell apoptosis, and alleviated oxidative damage. It inhibited ferroptosis by decreasing Fe2 +, ROS, MDA, and LPO levels, while upregulating SOD, GSH, GCL, and GPX4. Mechanistically, nuclear factor erythroid 2-related factor 2 (NRF2) was identified as a key responsive target through target prediction and qPCR validation, while molecular docking and microscale thermophoresis (MST) assays demonstrated a direct interaction between Notoginsenoside R1 and SQSTM1 (p62). Besides, Notoginsenoside R1 enhanced p62 expression and promoted NRF2 nuclear translocation, restoring expression of GCLC, GCLM, and GPX4. Both p62 knockout and siRNA-mediated knockdown progressively attenuated these protective effects. These findings suggest that Notoginsenoside R1 protects against SSNHL by inhibiting ferroptosis and oxidative stress via activation of the p62-NRF2-GCL-GPX4 axis.

Ethnopharmacological relevance : Dihuang Yinzi (DY) is a classic formula traditionally used for stroke-related disorders. Its potential therapeutic effect on post-stroke depression (PSD), however, remains to be investigated. Aim of the study : This study aims to evaluate the antidepressant effects of DY in a rat model of PSD and to explore the underlying mechanisms involving mitophagy and neuroinflammation. Materials and methods PSD was induced in rats by middle cerebral artery occlusion combined with chronic unpredictable mild stress. DY extract was administered daily for two weeks. Neurofunctional deficits and depressive-like behaviors were assessed. Cerebral infarction volume was measured by TTC staining. Hippocampal tissues were collected for transcriptomic analysis, qPCR, Western blot, immunoprecipitation, and ELISA to detect mitophagy-related genes (RHOT1, MAP1LC3B, PINK1, PRKN) and proteins as well as inflammatory factors. A specific mitophagy inhibitor was used to verify whether the anti-inflammatory effects of DY depend on mitophagy. Subcellular fractionation and immunofluorescence co-localization with the mitochondrial marker Tom20 were performed to confirm mitochondrial localization of overexpressed Miro1. The role of Miro1 was further validated using adenovirus-mediated overexpression. Results DY significantly alleviated depressive-like behaviors and reduced cerebral infarction volume in PSD rats. qPCR analysis showed that DY downregulated RHOT1 (Miro1) and upregulated MAP1LC3B, PINK1, and PRKN mRNA expression. Transcriptomic and mechanistic analyses revealed that DY promoted Miro1 degradation, thereby enhancing PINK1-mediated mitophagy. Subcellular fractionation and immunofluorescence confirmed that overexpressed Miro1 was correctly localized to mitochondria. Inhibition of mitophagy abolished the anti-inflammatory effects of DY, as evidenced by increased NF-κB activation and elevated pro-inflammatory cytokine levels. Notably, adenovirus-mediated Miro1 overexpression reversed the therapeutic effects of DY. Conclusions :DY improves PSD by promoting Miro1 degradation to facilitate mitophagy and suppress NF-κB-mediated neuroinflammation. These findings provide a scientific basis for the clinical application of DY in PSD and identify Miro1 as a potential therapeutic target.

Vascular dementia (VaD) is a neurodegenerative disease primarily characterized by white matter injury and myelin degeneration, and currently, there is a lack of effective treatment options. This study aims to investigate the effects of the traditional Chinese medicine formula Danggui Shaoyao San (DSS) on cognitive function and myelin repair in VaD rats and to elucidate its underlying mechanisms. The VaD rat model was established using the bilateral common carotid artery ligation (2VO) method. The effects of DSS on cognitive function, myelin regeneration, sphingolipid metabolism, and SPHK2/S1P/S1PR5 pathway was conducted using behavioral tests, histological staining, Western blot, lipidomics, qPCR, immunofluorescence, LC–MS/MS, and 16S rRNA sequencing. Besides, molecular docking and molecular dynamics simulation were carried out. DSS treatment significantly improved learning and memory abilities in VaD rats, reduced structural damage in the hippocampus and white matter, and promoted the differentiation of oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes (OLs). Lipidomics and molecular biological experiments indicated that DSS activated the SPHK2/S1P/S1PR5 pathway, ameliorated sphingolipid metabolic disorders and increased S1P levels, thereby promoting myelin repair. The specific SPHK2 inhibitor ABC294640 significantly weakened the neuroprotective effects of DSS, further confirming the central role of SPHK2/S1P/S1PR5 pathway. Antibiotic depletion experiments confirmed that the gut microbiota was not a key mediator of the therapeutic effects of DSS. Finally, molecular docking and molecular dynamics simulations indicated that the DSS components Albiflorin and Gallic acid form tighter and more stable interactions with SPHK2. DSS improved VaD cognitive impairment by modulating sphingolipid metabolism and promote myelin regeneration via activating the SPHK2/S1P/S1PR5 signaling pathway. This study provides important experimental evidence for the clinical application of DSS in VaD.

Diabetic wound healing remains a significant clinical challenge, characterized by a protracted and uncertain prognosis. Extracellular vesicles (EVs), functioning as natural carriers released by living cells, play a pivotal role in intercellular communications by delivering diverse bioactive cargo. In recent years, plant-derived extracellular vesicles (PDEVs) have garnered increasing attention due to their inherent biocompatibility, safety, low immunogenicity, and abundant source availability. PDEVs are regarded as a highly promising cell-free therapeutic strategy for diabetic wound healing. This review systematically summarizes the research progress on PDEVs biogenesis, physiological functions and their underlying mechanisms, and isolation/characterization methodologies. Specifically, we explore the potential of PDEVs as drug delivery vehicles and discuss engineering strategies for their modification. Finally, we provide a critical analysis of the potential challenges associated with translating PDEVs into cell-free therapeutics for diabetic wounds and offer perspectives on future research directions.

ETHNOPHARMACOLOGICAL RELEVANCE:Glycyrrhiza uralensis Fisch. (GU) is a pivotal botanical in traditional Chinese medicine (TCM), because of its ability to reconcile various herbs, and its crucial role in numerous formulas. Huanggui Shaogan decoction (HSGD) is an empirical formula, developed by Huangqi Guizhi Wuwu decoction (HGD) and Shaoyao Gancao decoction (SGD), and prepared by adding GU to HGD. However, the mechanisms of GU reconciling medicine remain incompletely characterized. AIM OF THE STUDY:This study aimed to explain mechanisms of GU reconciling medicine based on the differences of components, pharmacological efficacy, and the existence of the components. METHODS:Differential components between HSGD and HGD were systematically identified using LC‒MS/MS coupled with chemometric analysis. The existence states and binding affinities of these differential components were further characterized via ultrafiltration separation. The therapeutic potential of HSGD was validated in a murine model of oxaliplatin-induced peripheral neuropathy (OIPN). RESULTS:Twenty-two differential chemical components between HSGD and HGD were identified, including flavonoids, saponins, gingerol, and monoglycoside. The transmittance of flavonoids and gingerols increased in HSGD. However, the transmittance of astragalus saponins decreased, which may be due to micelle association and the increase in molecular clusters. HSGD could enhance the mechanical pain threshold, alleviate cold nociceptive hypersensitivity, relieve dorsal root ganglia neuron injury, and decrease the expression of nerve growth factor, 5-hydroxytryptamine, substance P, and calcitonin gene-related peptide better. The differential correlation analysis revealed the relationship between differential components and pharmacological indicators. The above results indicated that different herbs combinations had a greater impact on the dissolution and molecular state of the components of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. CONCLUSION:The study highlights the solubilizing effect of GU within HSGD, and it also improves the efficacy of the treatment of OIPN, which underpins its compatibility rationality. It provided an inspiration for the study of other TCM formulas.

Background:Depression and anxiety are significant global health concerns, with systemic inflammation playing a critical role in their pathophysiology. Recent studies have highlighted the C-reactive protein to lymphocyte ratio (CLR) as a potential biomarker of inflammation that may be associated with these mental health conditions. However, the relationship between CLR and depression and anxiety, especially within a diverse population, remains underexplored. Methods:This study utilized data from the National Health and Nutrition Examination Survey (NHANES) (2015-2023) to examine the association between CLR and the prevalence of depression and anxiety. A total of 22,308 participants were included for depression analysis, and 16,138 participants were included for anxiety analysis. Depression was assessed using the PHQ-9, and anxiety was assessed through self-reported anxiety symptoms and medication use. CLR was calculated as the ratio of C-reactive protein to lymphocyte count, and logistic regression models were applied to analyze associations, adjusting for demographic and health-related variables. Results:Higher CLR levels were significantly associated with increased odds of depression (OR: 1.49; 95% CI: 1.25-1.78) and anxiety (OR: 1.13; 95% CI: 1.02-1.26) after full adjustment for confounders. Non-linear relationships were observed, with specific inflection points for both depression (CLR = 0.96) and anxiety (CLR = 0.88), beyond which the risk of mental health disorders increased sharply. Subgroup analyses revealed that younger individuals and those without hypertension showed stronger associations between CLR and depression. Conclusion:Elevated CLR is associated with an increased risk of depression and anxiety, suggesting the potential role of systemic inflammation in influencing mental health outcomes. CLR may serve as a useful biomarker for identifying populations at higher risk, underscoring the need for further research into early intervention strategies and targeted approaches to address systemic inflammation in mental health care.

ETHNOPHARMACOLOGICAL RELEVANCE:Huangqi Guizhi Wuwu decoction (HGWD) is firstly recorded in the ancient Chinese ethnomedical manuscript "Synopsis of the Golden Chamber" with nourishing qi and blood for the treatment of limb numbness and ache in patients. Nowadays, accumulating evidence suggests that it could effectively alleviate symptoms of pain in oxaliplatin-induced peripheral neuropathy (OIPN). However, the therapeutic mechanism of HGWD against OIPN is necessary to be further clarified. METHODS:The mice model of OIPN was established using the behavioral tests and histopathological observation to assess the therapeutic effect of HGWD against OIPN in vivo. The UHPLC-ESI-Q-TOF-MS/MS was used to identify the potential bioactive components from HGWD in drug-containing rat plasma. The integrated network pharmacology and transcriptomics strategy was performed to deciphers the mechanism of HGWD against OIPN with experimental verification. RESULTS:HGWD could improve pain sensitivity symptom and alleviate the damage of dorsal root ganglia (DRG) neurons or intraepidermal nerve fibers in the mice model of OIPN. Through the integrated analysis of transcriptome and network pharmacology, Sirtuin 1 (Sirt1) was considered as the vital target for HGWD against OIPN. Via virtual screening and the ND7 cell viability assay, Formononetin (FN) and Oxypaeoniflorin (OPRA) from HGWD showed preliminary neuroprotective effects. In addition, OPRA can reduce Sterile alpha and toll interleukin receptor motif-containing protein 1 (SARM1) levels, improve axonal degeneration, and alleviate OIPN through the Sirt1 pathway. CONCLUSION:HGWD could effectively alleviate the pain and nerve injury symptoms in mice with oxaliplatin-induced peripheral neuropathy. The therapeutic mechanism of HGWD against OIPN was relevant to the Sirt1 for the function of anti-oxidative stress in DRG. OPRA can reduce SARM1 levels, improve axonal degeneration, and alleviate OIPN through the Sirt1 pathway.

Ginger, a well-known spice plant, has been used widely in medicinal preparations for pain relief. However, little is known about its analgesic components and the underlying mechanism. Here, we ascertained, the efficacy of ginger ingredient 8-Shogaol (8S), on inflammatory pain and tolerance induced by morphine, and probed the role of TRPV1 in its analgesic action using genetic and electrophysiology approaches. Results showed that 8S effectively reduced nociceptive behaviors of mice elicited by chemical stimuli, noxious heat as well as inflammation, and antagonized morphine analgesic tolerance independent on opioid receptor function. Genetic deletion of TRPV1 significantly abolished 8S’ analgesia action. Further calcium imaging and patch-clamp recording showed that 8S could specifically activate TRPV1 in TRPV1-expressing HEK293T cells and dorsal root ganglion (DRG) neurons. The increase of [Ca2+]i in DRG was primarily mediated through TRPV1. Mutational and computation studies revealed the key binding sites for the interactions between 8S and TRPV1 included Leu515, Leu670, Ile573, Phe587, Tyr511, and Phe591. Further studies showed that TRPV1 activation evoked by 8S resulted in channel desensitization both in vitro and in vivo, as may be attributed to TRPV1 degradation or TRPV1 withdrawal from the cell surface. Collectively, this work provides the first evidence for the attractive analgesia of 8S in inflammatory pain and morphine analgesic tolerance mediated by targeting pain-sensing TRPV1 channel. 8S from dietary ginger has potential as a candidate drug for the treatment of inflammatory pain.

Ethnopharmacological relevance: Invigorating blood circulation to remove blood stasis is a primary strategy in TCM for treating vascular dementia (VaD). Danggui-Shaoyao San (DSS), as a traditional prescription for neuroprotective activity, has been proved to be effective in VaD treatment. However, its precise molecular mechanisms remain incompletely understood. Aim of the study: The specific mechanism underlying the therapeutic effects of DSS on VaD was explored by employing network pharmacology as well as in vivo and in viro experiment validation. Materials and methods: We downloaded components of DSS from the BATMAN-TCM database for target prediction. The intersection between the components of DSS and targets, PPI network, as well as GO and KEGG enrichment analysis were then performed. Subsequently, the potential mechanism of DSS predicted by network pharmacology was assessed and validated through VaD rat model induced by 2VO operation and CoCl2-treated PC12 cells. Briefly, the DSS extract were first quantified by HPLC. Secondly, the effect of DSS on VaD was studied using MWM test, HE staining and TUNEL assay. Finally, the molecular mechanism of DSS against VaD was validated by Western blot and RT-QPCR experiments. Results: Through network analysis, 137 active ingredients were obtained from DSS, and 67 potential targets associated with DSS and VaD were identified. GO and KEGG analysis indicated that the action of DSS on VaD primarily involves hypoxic terms and HIF-1 pathway. In vivo validation, cognitive impairment and neuron mortality were markedly ameliorated by DSS. Additionally, DSS significantly reduced the expression of proteins related to synaptic plasticity and neuron apoptosis including PSD-95, SYP, Caspase-3 and BCL-2. Mechanistically, we confirmed DSS positively modulated the expression of HIF-1 alpha and its downstream proteins including EPO, pEPOR, STAT5, EPOR, and AKT1 in the hippocampus of VaD rats as well as CoCl2-induced PC12 cells. HIF-1 inhibitor YC-1 significantly diminished the protection of DSS on CoCl2-induced PC12 cell damage, with decreased HIF-1 alpha, EPO, EPOR expression. Conclusion: Our results initially demonstrated DSS could exert neuroprotective effects in VaD. The pharmacological mechanism of DSS may be related to its positive regulation on HIF-1 alpha/EPO pathway.

Background Oxidative stress, imbalance of neurotransmitters and multidrug resistant transporters play pivotal roles in the development and treatment of seizures. Classical formula Baijin Pills (BJP) has been used to treat brain damage caused by epilepsy for thousands of years. However, little information on the anti-seizure effects of BJP has been reported so far. Purpose To investigate the effects of BJP on Lithium-pilocarpine- induced seizures and its role in the regulation of oxidative stress and neurotransmission. Study Design The present study elucidates the anti-epileptic effects and mechanisms of action of BJP, using a mouse model of Lithium-pilocarpine- induced seizures. Methods The antiepileptic effect of BJP was tested by fear conditioning test of mice model. Nissl staining showed neural loss and apoptosis in the hippocampus seizure mice and BJP treatment. Immunohistochemistry analysis of Bcl-2 and Bax inflected anti-apoptotic effects of BJP. The chemical colorimetric method was utilized to determine the effects of BJP on oxidative stress. The levels of excitatory and inhibitory neurotransmitters in cerebrospinal fluid (CSF) were detected by ELISA. Whole-cell votage-clamp recording was made to detect currents of receptor gated channels. Results We found that BJP can significantly improve the cognitive disorders of epileptic mice, prolonged the latency of seizures and reduced the duration of epilepsy. BJP also effectively ameliorate neural injury in the hippocampus, up-regulated Bcl-2 protein level whereas decreased Bax expression. Moreover, the levels of SOD, GSH as well as γ-aminobutyric acid (GABA) were increased, while the levels of MDA, aminomethyl phosphonic acid (AMPA) and N-Methyl-D-aspartic acid (NMDA) were decreased in the hippocampus with BJP treatment. Patch clamp recording showed that BJP had no significant effect on the current induced by GABA, NMDA and AMPA in acutely isolated mice hippocampal neurons. Furthermore, BJP inhibited the expression of P-gp, MRP1 and MDR1 protein. Conclusions Our study provides the first experimental basis that BJP can exert anti-seizure effects by attenuating oxidative stress, regulating neurotransmission and inhibit drug-resistant associated proteins. BJP may be developed as a drug candidate to treat epilepsy in clinical practice.

Chronic kidney diseases (CKD) is a serious threat to people's health with renal fibrosis as the major pathological feature. The absent in melanoma 2 (AIM2) has recently been proposed to play a critical role in CKD. Emodin is a major bioactive compound from rhubarb, which is widely used for clinical treatment of renal disease. The aim of this study is to elucidate the effect of emodin on unilateral ureteral obstruction (UUO) model mice and its association with the AIM2 inflammasome. In this study, we established the UUO-induced mice renal interstitial fibrosis in vivo and bone marrow-derived macrophages (BMDMs) model in vitro. The BUN, SCr, TNF-α, IL-1β in serum were examined. The degree of renal damage and fibrosis were determined by histological assessment. Immunofluorescence, western blot, and Co-IP were used to determine the mechanisms of emodin against CKD. Emodin could improve UUO-induced abnormal renal function and histopathological abnormalities. It could also ameliorate renal fibrosis, evidenced by inhibiting the expression of α-SMA, TGF-β1, FN, and collagen I. Mechanistically, emodin significantly suppressed AIM2 inflammasome as well as its components including ASC, cleaved caspase-1, and IL-1β both in vivo and in vitro. Further studies demonstrated that emodin inhibited K27-linked polyubiquitination of AIM2 by targeting on K64 sites of the lysine residues. In summary, emodin could hinder the activation of AIM2 inflammasome in UUO model mice through K27-linked polyubiquitination to reduce renal fibrosis. Emodin is a possible therapeutic option for CKD treatment.

Background: Dandouchi polypeptide (DDCP) is derived from Semen Sojae Praeparatum (Dandouchi in Chinese), a fermented product of Glycine max (L.) Merr. Semen Sojae Praeparatum is widely used in the food industry for its unique flavor and nutritional value, and DDCP, as its derivative, also shows potential health benefits in food applications. However, the specific active substances responsible for Semen Sojae Praeparatum and the underlying mechanisms involved have not been fully elucidated. Methods: DDCP was extracted from Semen Sojae Praeparatum using enzymes, and its antidepressant effects were tested in chronic unpredictable mild stress (CUMS)-induced mice. Immunohistochemistry, immunofluorescence, and western blotting were used to analyze neurogenesis and the nuclear factor kappa B (NF-kappa B) pathway. Moreover, an adeno-associated virus (AAV) shRNA was used to induce tripartite motif-containing 67 (TRIM67) deficiency to examine the function of TRIM67 in the neuroprotective effects of DDCP in depressive disorders. Results: DDCP reduced depressive behaviors in CUMS mice and the expression of proinflammatory markers in the hippocampus. DDCP promoted neurogenesis and modulated the TRIM67/NF-kappa B pathway, with TRIM67 deficiency impairing its antidepressant effect. Conclusions: This research revealed that DDCP has a protective effect on countering depression triggered by CUMS. Notably, TRIM67 plays a crucial role in mitigating depression through DDCP, positioning DDCP as a potential therapeutic option for treating depressive disorders.

PURPOSE Paclitaxel-induced cognitive impairment (PICI) is a frequent and severe adverse reaction of chemotherapy. Increased neuroinflammation related to neutrophil extracellular traps (NETs) may play a key role in PICI. Shaoyao Gancao Decoction (SGD) is a classic Chinese prescription with good neuroprotective activities. However, the action of SGD in PICI remains elusive. Herein, our work was set out to study the potential role and possible mechanism of SGD in PICI through network pharmacology, Mendelian analysis and animal experiments. METHODS First, network pharmacology analysis was performed to identify the potential active compounds and targets of SGD on PICI. Secondly, PICI mice model was established, and the neuroprotective effects of SGD on PICI mice were eveluated by using new object recognition, Morris water maze test and TUNEL staining. The impact of SGD on inflammatory factors, microglia activation and NETs formation were also studied by ELISA and immunofluorescence. The regulation of SGD on PTEN/PI3K/AKT pathway was detected by Western blot. Finally, Mendelian randomization was performed using genome-wide association analysis data of PTEN and cognitive disorders in OpenGWAS database. RESULTS Network pharmacological analysis revealed 105 active ingredients in SGD, and identified 124 targets associated with SGD and PICI. KEGG pathway analysis implied that PI3K-AKT signaling pathway maight be involved in the protective effect of SGD agianst PICI Animal experiments displayed that SGD significantly alleviated cognitive impairment induced by paclitaxel, and decreased neuronal apoptosis. Furthermore, SGD remarkably reduced IL-1β, TNF-α and microglia activation, while enhanced SOD activity in hippocampus. In addition, SGD could inhibit NETs formation, evidenced by decreasing the level of plasma MPO-DNA complex, as well as MPO and PAD4 expression. Western blotting assay suggested that SGD attenuated the elevated expression of PI3K, AKT, p-AKT and PKC-α induced by paclitaxel, whereas decreased PTEN expression. The results of Mendelian randomization analysis further confirm that PTEN is an important therapeutic target for improving cognitive function. CONCLUSION Collectively, our preliminary findings indicated that SGD can improve cognitive function of PICI mice. SGD may play a neuroprotective role by inhibiting NETs formation via PTEN/PI3K/AKT signaling pathway. SGD may be a potential alternative therapy for PICI.

The pathogenesis of coronary heart disease is a highly complex process, with lipid metabolism disorders being closely linked to its development. Therefore, this paper analyzes the various factors that influence lipid metabolism, including obesity, genes, intestinal microflora, and ferroptosis, through a comprehensive review of basic and clinical studies. Additionally, this paper delves deeply into the pathways and patterns of coronary heart disease. Based on these findings, it proposes various intervention pathways and therapeutic methods, such as the regulation of lipoprotein enzymes, lipid metabolites, and lipoprotein regulatory factors, as well as the modulation of intestinal microflora and the inhibition of ferroptosis. Ultimately, this paper aims to offer new ideas for the prevention and treatment of coronary heart disease.

目的 建立加味黄芪桂枝五物汤(Jiawei Huangqi Guizhi Wuwu Decoction,JHD)HPLC指纹图谱,鉴定其主要化学成分,并阐明指纹图谱共有峰的成分归属.方法 HPLC指纹图谱检测采用Hypersil ODS色谱柱(250 mm×4.6 mm,5 μm);流动相为乙腈-0.2％甲酸水溶液,梯度洗脱,体积流量1mL/min,检测波长为250nm,柱温为30℃;运用"中药色谱指纹图谱相似度评价系统"进行相似度评价,并以高效液相色谱串联四级杆飞行时间质谱(liquid chromatography-quadrupole time-of-flight mass spectrometry,LC-Q-TOF/MS)方法对指纹图谱中化学成分进行分析,通过对照品、化合物质谱信息结合文献报道鉴定其结构,进一步通过药材指纹图谱阐明共有峰的成分归属.结果 建立了 JHD的HPLC指纹图谱,确定了 20个共有峰,10批样品相似度为0.920～0.990.在此基础上共鉴定出JHD中70个化学成分,包括28个黄酮类、23个三萜类、5个单萜苷类、5个有机酸类、5个姜辣素类及4个其他类化合物.结论 HPLC指纹图谱结合LC-Q-TOF/MS定性鉴别,能够体现JHD组分的整体特征,可为其质量控制研究奠定基础.

Cerebral infarction is one of the most common diseases for aged people. Compound Tongluo Decoction (CTLD), a classic traditional Chinese Medicine prescription, has been widely used in the treatment of ischemic cerebral infarction. Transient middle cerebral artery occlusion (tMCAO) rat model is established for the animal experiment and oxygen-glucose deprivation and reperfusion (OGD/R) human umbilical vein endothelial cells (HUVECs) model are established for the cell experiment. This also use Nrf2-/- rats to detect the role of nuclear factor erythroid 2-related factor 2 (Nrf2). Longa score, Evans blue staining, brain water content measurement, and histological observation are done. The levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and other ferroptosis-related components are detected respectively. In the vivo experiment, CTLD relieved ischemia-reperfusion (IR) injury symptoms and attenuated IR injury in brain tissues of tMCAO rats by relieving peroxidation injury in brain tissues and inhibiting ferroptosis in tMCAO rats. Moreover, CTLD reversed OGD/R-induced oxidative damage of endothelial cells via suppressing ferroptosis. After knocking out the Nrf2 gene, the protective effect of CTLD is sharply reduced. This study put forward that CTLD can inhibit ferroptosis in I/R-injured vascular endothelium by regulating Nrf2/ARE/SLC7A11 signaling to improve the relative symptoms of rats after cerebral I/R injury, thus providing a viable treatment option for cerebrovascular disease.

The development of natural membranes as coatings for nanoparticles to traverse the blood-brain barrier (BBB) presents an effective approach for treating central nervous system (CNS) disorders. In this study, we have designed a nanogel loaded with PACAP and estrogen (E2), sheathed with exosomes and responsive to reactive oxygen species (ROS), denoted as HA NGs@exosomes. The objective of this novel design is to serve as a potent drug carrier for the targeted treatment of perimenopausal depression. The efficient cellular uptake and BBB penetration of HA NGs@exosomes has been demonstrated in vitro and in vivo. Following intranasal intervention with HA NGs@exosomes, ovariectomized mice under chronic unpredictable mild stress (CUMS) have shown improved behavioral performance, indicating that HA NGs@exosomes produced a rapid-onset antidepressant effect. Moreover, HA NGs@exosomes exhibit notable antioxidant and anti-inflammatory properties and may regulate the expression of pivotal proteins in the PACAP/PAC1 pathway to promote synaptic plasticity. Our results serve as a proof-of-concept for the utility of exosome-sheathed ROS-responsive nanogel as a promising drug carrier for the treatment of perimenopausal depression.

Background: Zhi-Zi-Chi-Tang (ZZCT) is an effective traditional Chinese medicinal formula. ZZCT has been used for the treatment of depression for centuries. Its clinical efficacy in relieving depression has been confirmed. However, the molecular mechanisms of ZZCT regarding neuroplasticity in the pathogenesis of depression have not yet been elucidated. Purpose: The present study aimed to examine the effects of ZZCT on neuroplasticity in mice exposed to chronic unpredictable mild stress (CUMS), and to explore the underlying molecular mechanisms. Methods: For this purpose, a murine model of depression was established using the CUMS procedure. Following the intragastric administration of ZZCT or fluoxetine, classic behavioral experiments were performed to observe the efficacy of ZZCT as an antidepressant. Immunofluorescence was used to label and quantify microtubule-associated protein (MAP2) and postsynaptic density protein (PSD95) in the hippocampus. Golgi staining was applied to visualize the dendritic spine density of neurons in the hippocampi. Isolated hippocampal slices were prepared to induce long-term potentiation (LTP) in the CA1 area. The hippocampal protein expression levels of glycogen synthase kinase-3 beta (GSK-3 beta), p-GSK-3 beta (Ser9), cAMP response element binding protein (CREB), p-CREB (Ser133), brain-derived neurotrophic factor (BDNF) and 14-3-3 zeta were detected using western blot analysis. The interaction of 14-3-3 zeta and p-GSK-3 beta (Ser9) was examined using co-immunoprecipitation. LV-shRNA was used to knockdown 14-3-3 zeta by an intracerebroventricular injection. Results: ZZCT (6 g/kg) and fluoxetine (20 mg/kg) alleviated depressive-like behavior, restored hippocampal MAP2(+) PSD95(+) intensity, and reversed the dendritic spine density of hippocampal neurons and LTP in the CA1 region of mice exposed to CUMS. Both low and high doses of ZZCT (3 and 6 g/kg) significantly promoted the binding of 14-3-3 zeta to p-GSK-3 beta (Ser9) in the hippocampus, and ZZCT (6 g/kg) significantly promoted the phosphorylation of GSK-3 beta Ser9 and CREB Ser133 in the hippocampus. ZZCT (3 and 6 g/kg) upregulated hip-pocampal BDNF expression in mice exposed to CUMS. LV-sh14-3-3 zeta reduced the antidepressant effects of ZZCT. Conclusion: ZZCT exerted antidepressant effects against CUMS-stimulated depressive-like behavior mice. The knockdown of 14-3-3 zeta using lentivirus confirmed that 14-3-3 zeta was involved in the ZZCT-mediated antide-pressant effects through GSK-3 beta/CREB/BDNF signaling. On the whole, these results suggest that the antide-pressant effects of ZZCT are attributed to restoring damage by neuroplasticity enhancement via the 14-3-3 zeta/ GSK-3 beta/CREB/BDNF signaling pathway.