1.Mechanism of Action of Kaixinsan in Ameliorating Alzheimer's Disease
Xiaoming HE ; Xiaotong WANG ; Dongyu MIN ; Xinxin WANG ; Meijia CHENG ; Yongming LIU ; Yetao JU ; Yali YANG ; Changbin YUAN ; Changyang YU ; Li ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):20-29
ObjectiveTo investigate the mechanism of action of Kaixinsan in the treatment of Alzheimer's disease (AD) based on network pharmacology, molecular docking, and animal experimental validation. MethodsThe Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and the Encyclopedia of Traditional Chinese Medicine(ETCM) databases were used to obtain the active ingredients and targets of Kaixinsan. GeneCards, Online Mendelian Inheritance in Man(OMIM), TTD, PharmGKB, and DrugBank databases were used to obtain the relevant targets of AD. The intersection (common targets) of the active ingredient targets of Kaixinsan and the relevant targets of AD was taken, and the network interaction analysis of the common targets was carried out in the STRING database to construct a protein-protein interaction(PPI) network. The CytoNCA plugin within Cytoscape was used to screen out the core targets, and the Metascape platform was used to perform gene ontology(GO) functional enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis. The “drug-active ingredient-target” interaction network was constructed with the help of Cytoscape 3.8.2, and AutoDock Vina was used for molecular docking. Scopolamine (SCOP) was utilized for modeling and injected intraperitoneally once daily. Thirty-two male C57/BL6 mice were randomly divided into blank control (CON) group (0.9% NaCl, n=8), model (SCOP) group (3 mg·kg-1·d-1, n=8), positive control group (3 mg·kg-1·d-1 of SCOP+3 mg·kg-1·d-1 of Donepezil, n=8), and Kaixinsan group (3 mg·kg-1·d-1 of SCOP+6.5 g·kg-1·d-1 of Kaixinsan, n=8). Mice in each group were administered with 0.9% NaCl, Kaixinsan, or Donepezil by gavage twice a day for 14 days. Morris water maze experiment was used to observe the learning memory ability of mice. Hematoxylin-eosin (HE) staining method was used to observe the pathological changes in the CA1 area of the mouse hippocampus. Enzyme linked immunosorbent assay(ELISA) was used to determine the serum acetylcholine (ACh) and acetylcholinesterase (AChE) contents of mice. Western blot method was used to detect the protein expression levels of signal transducer and activator of transcription 3(STAT3) and nuclear transcription factor(NF)-κB p65 in the hippocampus of mice. ResultsA total of 73 active ingredients of Kaixinsan were obtained, and 578 potential targets (common targets) of Kaixinsan for the treatment of AD were screened out. Key active ingredients included kaempferol, gijugliflozin, etc.. Potential core targets were STAT3, NF-κB p65, et al. GO functional enrichment analysis obtained 3 124 biological functions, 254 cellular building blocks, and 461 molecular functions. KEGG pathway enrichment obtained 248 pathways, mainly involving cancer-related pathways, TRP pathway, cyclic adenosine monophosphate(cAMP) pathway, and NF-κB pathway. Molecular docking showed that the binding of the key active ingredients to the target targets was more stable. Morris water maze experiment indicated that Kaixinsan could improve the learning memory ability of SCOP-induced mice. HE staining and ELISA results showed that Kaixinsan had an ameliorating effect on central nerve injury in mice. Western blot test indicated that Kaixinsan had a down-regulating effect on the levels of NF-κB p65 phosphorylation and STAT3 phosphorylation in the hippocampal tissue of mice in the SCOP model. ConclusionKaixinsan can improve the cognitive impairment function in SCOP model mice and may reduce hippocampal neuronal damage and thus play a therapeutic role in the treatment of AD by regulating NF-κB p65, STAT3, and other targets involved in the NF-κB signaling pathway.
2.Molecular expression and pathological morphologic changes of extraocular muscle in concomitant exotropia
Xiaorui ZHOU ; Zhibin WANG ; Yu DI
International Eye Science 2025;25(1):55-58
Strabismus, a common ocular condition, arises from an imbalance in the extraocular muscle force and deviation of the visual axis due to various factors. Concomitant strabismus is the predominant form of exotropia, with its pathogenesis believed to be associated with hereditary factors, abnormal eye accommodation function, and anomalies in binocular anatomy. Surgical intervention is often necessary for aligning the visual axes of both eyes and facilitating the recovery and establishment of stereoscopic vision. Despite this, the etiology of concomitant exotropia remains incompletely understood. This review consolidates recent research on aberrant molecular expression and pathological morphological changes within extraocular muscles affected by concomitant exotropia, offering insights into disease causation at molecular and pathological levels to underpin future preventive measures and clinical interventions. The discussion encompasses histological changes observed under electron microscopy as well as the impact of heavy chain protein, satellite cells, cadherin, growth factors on extraocular muscle protein expression.
3.Textual Research on Key Information and Modern Clinical Application of Classical Famous Formula Liumotang
Xinyu ZHANG ; Chong LI ; Yixuan HU ; Luming LIANG ; Ye ZHAO ; Xiaoting LU ; Yu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):201-212
Liumotang comes from the Yuan dynasty's Effective Prescription Handed Down for Generations of Physicians. It is composed of six medicinal materials: Arecae Semen, Aquilariae Lignum Resinatum, Aucklandiae Radix, Linderae Radix, Rhei Radix et Rhizoma, and Aurantii Fructus. It is a classical formula for treating abdominal pain due to Qi stagnation and constipation accompanied by heat. This study systematically collated the records of Liumotang in ancient medical books and modern clinical literature and conducted in-depth analysis and textual research on its formula source, main diseases, composition, dosage, medical books, container capacity, processing, preparation method, usage, drug basis, formula meaning, and other key information, so as to provide a powerful reference for the development and clinical application of compound preparations of the classical formula Liumotang. The results show that Liumotang was first seen in Effective Prescription Handed Down for Generations of Physicians, and many medical books of the past dynasties have imitated this. In terms of drug basis, the dried and mature seeds of the palm plant Areca catechu, resin-containing wood of the Daphneaceae plant Aquilaria sinensis, the dried roots of the Asteraceae plant woody Aucklandia lappa, the dried tuber root of the Lauraceae plant Lindera aggregata, the dried roots and rhizomes of the knotweed plant, R. palmatum, R.tangutikum, and R. officinale, and the dried and unripe fruits of the citrus genus C. aurantium and its cultivated varieties from the family Rutaceae were selected. In terms of dosage, through the textual research on bowls in the Ming and Qing dynasties, combined with the conversion of medicines and bowl capacity in the Qing dynasty, it was estimated that the dosage of each drug in the Yuan dynasty was 10.86 g. In the Ming and Qing dynasties, the dosage of drugs was mostly equal, but the dosage of drugs was somewhat different. In terms of processing, preparation method, and usage, in the medical books of the past dynasties, the processing of drugs has slightly changed, but raw drugs are used in all preparations. The preparation method and usage did not change much during the Yuan, Ming, and Qing dynasties, except for certain differences in dosage. In terms of syndrome, Liumotang was first used to treat abdominal pain due to Qi stagnation and constipation accompanied by heat. Medical books of the past dynasties often omit the symptoms of heat. In modern clinical practice, Liumotang is mainly used in the digestive system and urinary system diseases and is mostly used to treat constipation-predominant irritable bowel syndrome, biliary reflux gastritis, functional constipation, slow transit constipation, and other diseases, with no adverse reactions found yet. The above results provide a reliable scientific basis for the development and clinical treatment of Liumotang compound preparations.
4.Analysis of Application of Animal Model of Spleen Deficiency and Dampness Syndrome Based on Data Mining
Qingqian YU ; Yifei ZHANG ; Zehan ZHANG ; Weiyue ZHANG ; Yuebo WANG ; Fengzhi WU ; Feng LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):235-243
ObjectiveThe research focuses on developing modeling and evaluation methodologies for an animal model exhibiting spleen deficiency and dampness excess syndrome, with the aim of standardizing such animal models for future reference. MethodsBy conducting a literature search on animal models of spleen deficiency and dampness excess syndrome, relevant publications meeting inclusion and exclusion criteria will be identified based on publication date, data source, types of diseases involved, animal characteristics, modeling methods, modeling duration, macroscopic syndrome assessment indicators, macroscopic quantification indicators, laboratory testing parameters, intervention approaches, positive controls and application context. A database will be established to facilitate the extraction of this information for quantitative analysis, statistical evaluation, and visual representation. ResultsA total of 137 literature articles meeting the standards have been included in the research. The primary animal species used in animal models of spleen deficiency and dampness excess are SD rats. Modeling methods include single-factor, dual-factor composite, and triple-factor composite methods, with various models widely applied in validation of pharmacological effects and mechanistic explorations. Evaluation indices of animal models for spleen deficiency and dampness excess primarily consist of macroscopic syndrome evaluation indicators and macroscopic quantitative indicators. Laboratory testing indicators are mostly related to research areas such as fluid metabolism and gastrointestinal function. The most commonly studied herbal formulas currently include Shenling Baizhu San and Pingwei San, with natural recovery and the use of the western medicine metronidazole as the most frequently used positive controls. ConclusionThe application of animal models for spleen deficiency and dampness excess is gradually increasing, with various modeling methods already simulating the typical characteristics of this syndrome pattern. However, there are still many areas that are worth contemplating and improving. This study aims to provide reference and ideas for the standardization of symptom names in animal models of spleen deficiency and dampness excess, as well as for the improvement of model construction and evaluation systems.
5.Effects of borneol on pharmacodynamics and pharmacokinetics of Corydalis saxicola total alkaloids in depression model rats
Yu YE ; Guoliang DAI ; Huaxi HANG ; Meishuang YU ; Yiran WANG ; Xuewen SHAO ; Wenzheng JU
China Pharmacy 2025;36(1):30-36
OBJECTIVE To investigate the effects of borneol on pharmacodynamic and pharmacokinetic effects of Corydalis saxicola total alkaloids in depression model rats. METHODS Thirty male SD rats were divided into blank control group, negative control group, positive control group (fluoxetine 10 mg/kg, i.g.), single drug group (C. saxicola total alkaloids 210 mg/kg, i.g.) and combined drug group (C. saxicola total alkaloids 210 mg/kg+borneol 50 mg/kg, i.g.) according to the random number table method, with 6 rats in each group. By lipopolysaccharide (LPS) induction modeling, except blank control group (no model and no administration) received intraperitoneal injection of the same amount of normal saline, the rats in the other groups were intraperitoneally injected with LPS once a day to establish a rat model of depression. After 1 week of modeling, each administration group was given relevant drug intragastrically according to the corresponding dose, and blank control group and negative control group (without drug treatment) were administered intragastrically with an equal volume of solvent to dissolve the drug; continued modeling while administering the drug. After two weeks of continuous administration, the effects of C. saxicola total alkaloids versus the combination of C. saxicola total alkaloids and borneol on the behavior of depressed rats were tested by behavioral experiments; the levels of tumor necrosis factor-α, interleukin-1β and interleukin-6 in rats were determined; the histopathological changes of the hippocampus of rats were observed. Blood sample was collected from the orbit at different time points after administration on the 15th day, and the upper plasma was obtained. Ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry was established for the simultaneous determination of dehydrocarvedine, tetrahydropalmatine, coptisine, palmatine, jatrorrhizine, berberine, berberrubine and epiberberine in rat plasma. The average plasma concentration-time curve was depicted, the area under the curve (AUC) was calculated, and the pharmacokinetic parameters were analyzed by DAS 3.2.2 software. RESULTS Compared with blank control group, the negative control group had a decrease in body mass and sugar water preference rate, a decrease in the total distance of open field, a prolonged swimming immobility time, and a increased in the expression of inflammatory factors in serum (P<0.05); compared with negative control group, the single drug group and the combined drug group increased the preference rate of sugar water, increased the total distance of open field, shortened the time of swimming immobility, and decreased the expression of inflammatory factors in serum (P<0.05). There was no significant difference in the above indicators between the single drug group and the combined drug group in rats (P>0.05). Pharmacokinetic results showed that compared with single drug group, AUC0-t of coptisine, AUC0-t, AUC0-∞, tmax and cmax of jatrorrhizine, AUC0-t, AUC0-∞, t1/2 and cmax of berberrubine, and AUC0-t of epiberberine, cmax of dehydrocarvedine, cmax of palmatine were significantly increased in combined drug group, but there was no significant difference, indicating that borneol didn’t have a significant effect on the efficacy of Corydalis saxicola nigra at this dose. CONCLUSIONS Both C. saxicola total alkaloids alone and in combination with borneol can improve depression-like behavior in depression model rats, reduce serum inflammatory cytokine levels, and protect hippocampal neurons. Compared with the use of Corydalis saxicola base alone, the combination with borneol do not show significant pharmacodynamic differences, bu can improve the absorption of coptisine, jatrorrhizine in model rats.
6.Meta-analysis of the relationship of semaglutide and malignant neoplasms risk in type 2 diabetes mellitus patients
Qingchuan LIAO ; Wei YU ; Quan WANG
China Pharmacy 2025;36(1):117-123
OBJECTIVE To systematically evaluate the relationship of semaglutide with malignant neoplasms in type 2 diabetes mellitus (T2DM) patients. METHODS Retrieved from the Cochrane Library, PubMed, Embase, ClinicalTrials.gov, CNKI, Wanfang data and CBM, randomized controlled trials (RCTs) about semaglutide in the treatment of T2DM patients with outcome measures including malignant tumor events were collected from the establishment of the database to June 2024. Meta- analysis was performed by using RevMan 5.3 software to assess the risk of malignant neoplasms. RESULTS A total of 24 RCTs (26 trials) involving 24 145 patients were included. Results of meta-analysis showed that compared to placebo, there was no statistical significance in the risk of semaglutide in pancreatic cancer [RR=0.39, 95%CI(0.10, 1.50), P=0.17], thyroid cancer [RR=1.29, 95%CI(0.38, 4.36), P=0.68], prostate cancer [RR=1.05, 95%CI(0.36, 3.12), P=0.92], skin cancer [RR=1.27, 95%CI(0.80, 2.02), P=0.31], gastrointestinal cancer [RR=1.00, 95%CI(0.47, 2.14), P=1.00], colorectal cancer [RR=0.96, 95%CI(0.40, 2.26), P=0.92], lung cancer [RR=1.62, 95%CI(0.74, 3.55), P=0.23], breast cancer [RR=1.25, 95%CI(0.45, 3.51), P=0.67] or all malignant neoplasms [RR=0.96, 95%CI(0.76, 1.21), P=0.73]. Compared to other antidiabetic drugs, there was no statistical significance in the risk of semaglutide in pancreatic cancer [RR=0.62, 95%CI(0.18, 2.09), P=0.44], thyroid cancer [RR=1.09, 95%CI(0.25, 4.78), P=0.90], prostate cancer [RR=2.09, 95%CI(0.46, 9.47), P=0.34], skin cancer [RR=1.76, 95%CI(0.65, 4.72), P=0.26], gastrointestinal cancer [RR=0.68, 95%CI(0.19, 2.35), P=0.54], colorectal cancer [RR=0.60, 95%CI(0.20, 1.78), P=0.36], lung cancer [RR=1.00, 95%CI(0.24, 4.11), P=1.00], breast cancer [RR=0.82, 95%CI(0.25, 2.66), P=0.74] or all malignant neoplasms [RR=1.36, 95%CI(0.96, 1.94), P=0.09]. CONCLUSIONS Semaglutide does not increase the risk of any type of malignant neoplasms in T2DM patients.
7.Effect of Acupuncture Combined with Bloodletting and Cupping on the Expression of Coagulation-Complement-Mast Cell Activation Axis-Related Factors in Patients with Chronic Spontaneous Urticaria:Randomize-controlled Study
Yuzhu DU ; Yuqiang XUE ; Xiang LIU ; Yu SHI ; Hongkun LI ; Wenshan LIU ; Zan TIAN ; Yutong HU ; Yanjun WANG
Journal of Traditional Chinese Medicine 2025;66(2):150-156
ObjectiveTo observe the clinical efficacy of acupuncture combined with bloodletting and cupping in the treatment of chronic spontaneous urticaria(CSU) and to explore its potential mechanisms of action. MethodsSeventy CSU patients were randomly divided into loratadine group and acupuncture + bloodletting group, with 35 patients in each group. The loratadine group received oral loratadine tablets, 10 mg once daily in the evening. The acupuncture + bloodletting group received acupuncture at Zhongwan (CV 12), Guanyuan (CV 4), Tianshu (ST 25), Zusanli (ST 36), Sanyinjiao (SP 6), Xuehai (SP 10), Quchi (LI 11), Hegu (LI 4), Taichong (LR 3), Baihui (GV 20), and Shenting (GV 24), once daily,along with bloodletting and cupping at Dazhui (GV 14) and Geshu (BL 17), every other day. Both groups were treated for 4 weeks. The 7-day urticaria activity score(UAS7) was assessed before and after the treatment, and levels of serum immunoglobulin E (IgE), interleukin-4 (IL-4), interleukin-5 (IL-5), eosinophil cationic protein (ECP), plasma tissue factor (TF), activated factor Ⅶ (FⅦa), prothrombin fragment 1+2 (F1+2), D-dimer (D-D) and complement component 5a (C5a) were detected. ResultsA total of 65 patients were included in the final analysis, 32 in the loratadine group and 33 in the acupuncture + bloodletting group. Before treatment, there was no significant difference in UAS7 score, serum IgE, IL-4, IL-5, ECP levels, or plasma TF, FⅦa, F1+2, D-D, C5a levels between groups (P> 0.05). After treatment, both groups showed significant reductions in UAS7 score, serum IgE, IL-4, IL-5, and plasma TF, FⅦa, F1+2, D-D, and C5a levels compared to those before treatment (P<0.01). However, after treatment, there was no significant difference in UAS7 score and serum ECP, IgE, IL-4, IL-5 levels between groups (P>0.05). The acupuncture + bloodletting group showed lower plasma TF, FⅦa, F1+2, D-D and C5a levels compared to the loratadine group (P<0.05 or P<0.01). ConclusionAcupuncture combined with bloodletting and cupping can effectively improve the skin symptoms of CSU patients and reduce the levels of inflammatory factors. The potential mechanism of action may involve the regulation of the coagulation-complement-mast cell activation axis, thereby inhibiting mast cell degranulation.
8.Exploration of the Intervention Mechanism of Qingshi Anti-itch Ointment (青石止痒软膏) on Psoriasis Model Mice Based on Caspase-1/GSDMD Protein Pathway
Yatong LI ; Yuanwen LI ; Yutong DENG ; Xuewen REN ; Xuewan WANG ; Xinhui YU ; Tangyunni LIU
Journal of Traditional Chinese Medicine 2025;66(2):170-177
ObjectiveTo explore the possible action mechanism of Qingshi Anti-itch Ointment (青石止痒软膏, QAO) in the treatment of psoriasis. MethodsForty mice were randomly divided into four groups, blank group, model group, calcipotriol group and QAO group, with 10 mice in each group. Except for the blank group, psoriasis was induced by applying imiquimod cream to the dorsal skin. After modeling for 6 hours daily, the calcipotriol group and QAO group were treated with 0.5 g of calcipotriol ointment or 0.5 g of QAO, respectively, applied to the treated dorsal skin. The blank group and the model group received no treatment. The skin lesions were observed, and the psoriasis area and severity index (PASI) score was assessed every other day. After 7 days, Hematoxylin and Eosin (HE) staining was performed on dorsal skin tissue to observe pathological changes. The levels of interleukin 1β (IL-1β) and interleukin 18 (IL-18) were determined by enzym-linked immunosorbent assay (ELISA). The protein levels of Caspase-1,Pro-Caspase-1, gasdermin D (GSDMD) and gasdermin-D-N (GSDMD-N) were detected by Western Blot (WB). The protein levels of GSDMD were observed by immunohistochemistry. ResultsCompared with the blank group, the model group mice showed redness, erythema, and white scales on their skin, with histological observations indicating epidermal thickening, elongated spines, and infiltration of inflammatory cells. The PASI scores of the skin tissue on days 1, 3, 5, and 7 were elevated; the IOD and AOD values of GSDMD protein increased; the protein levels of Caspase-1, Pro-Caspase-1,GSDMD, GSDMD-N, and IL-1β and IL-18 were significantly elevated (P<0.05 or P<0.01). Compared with the model group, the QAO group and calcipotriol group showed lighter skin lesions; the PASI scores on day 5 and day 7 in the QAO group, and on day 3, 5, and 7 in the calcipotriol group, were reduced; the IOD and AOD values of GSDMD protein, and the protein level of Caspase-1, GSDMD, and GSDMD-N, as well as level of IL-18 and IL-1β decreased in both groups; in the calcipotriol group, Pro-Caspase-1 protein level also decreased (P<0.05 or P<0.01). Compared with the calcipotriol group, the QAO group showed slightly redder skin, more obvious thickening of the stratum corneum, and less capillary dilation; the PASI scores on day 3 and day 7 increased, while the score on day 5 was reduced; the protein level of Pro-Caspase-1, GSDMD, GSDMD-N, and the level of IL-18 and IL-1β were increased in the QAO group (P<0.05). ConclusionQAO can effectively relieve psoriasis dermatitis in mice. Its potential mechanism may be related to the regulation of the Caspase-1/GSDMD protein pathway, down-regulation of IL-18 and IL-1β levels, and alleviation of pyroptosis.
9.Availability and use of child safety seats among children aged 0-3 years
CHEN Bo ; WANG Xihui ; QIU Fengqian ; YU Yan ; GAO Shuna ; HE Lihua ; LI Weiyi ; JI Yunfang ; CHEN Weihua
Journal of Preventive Medicine 2025;37(1):21-25
Objective:
To investigate the availability and use of child safety seats among children aged 0-3 years, so as to provide the basis for improving riding safety for children.
Methods:
Parents of children aged 0-3 years in Huangpu District, Shanghai Municipality, were recruited using the stratified multistage random sampling method from May to July 2024. Demographic information, family travel patterns, the use of child safety seat and related health beliefs were collected using questionnaire surveys. Factors affecting the use of child safety seats were identified using a multivariable logistic regression model.
Results:
Totally 514 valid questionnaires were recovered, with an effective rate of 96.98%. The respondents included 122 fathers (23.74%) and 392 mothers (76.26%), with a median age of 34.00 (interquartile range, 5.00) years. There were 446 families equipping with child safety seats, accounting for 86.77%; and 169 families using child safety seats, accounting for 32.88%. Multivariable logistic regression analysis showed that the parents who had children aged >1-2 years (OR=0.597, 95%CI: 0.366-0.973), travelled 2-4 times per month (OR=0.359, 95%CI: 0.213-0.607) or once per month or less (OR=0.384, 95%CI: 0.202-0.729), and scored high in perceived barrier (OR=0.634, 95%CI: 0.486-0.827) were less likely to use child safety seats; the parents who had children with local household registration (OR=2.506, 95%CI: 1.356-4.633), travelled 5-<10 km (OR=1.887, 95%CI: 1.148-3.101) or ≥10 km (OR=2.319, 95%CI: 1.355-3.967), always wore seat belts (OR=2.342, 95%CI: 1.212-4.524), scored high in perceived susceptibility (OR=1.392, 95%CI: 1.091-1.778) and self-efficacy (OR=1.413, 95%CI: 1.156-1.727) were more likely to use child safety seats.
Conclusions
Equipping family cars with child safety seats and using them can prevent and reduce traffic injuries among children aged 0-3 years. It is recommended to strengthen publicity to promote the use of child safety seats.
10.Fufang Kangjiaolv Capsules Treat Anxiety in Rat Model of Chronic Restraint Stress via Microbiota-gut-brain Axis
Wenxin FAN ; Tingyue JIANG ; Yu WANG ; Ge ZHANG ; Yifan LU ; Mengmeng LIU ; Jiayuan LI ; Renzhi MA ; Jinli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):95-107
ObjectiveTo observe the intervention effect of Fufang Kangjiaolv capsules on anxiety-like behaviors in the rat model of chronic restraint stress (CRS) and explore the mechanism underlying the anti-anxiety effect via the microbiota-gut-brain axis. MethodsRats were assigned into blank, model, positive drug (diazepam, 1 mg·kg-1), and low-, medium-, and high-dose (0.75, 1.5, 3 g·kg-1, respectively) Fufang Kangjiaolv capsules groups. After 14 days of administration, the elevated plus maze test, open field test, light and dark box test, and marble burying test were performed. Hematoxylin-eosin staining was employed to observe the pathological changes in the hippocampus and colon of rats, and Nissl staining was conducted to observe the damage of hippocampal neurons. The gut microbiota was analyzed by 16S rRNA gene sequencing. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA levels of zonula occludens-1 (ZO-1) and occludin in the colon of rats. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in the colon, serum, and hippocampus were determined by enzyme-linked immunosorbent assay. Western blot was employed to determine the protein levels of ZO-1, occludin, nuclear factor-κB p65 (NF-κB p65) in the colon tissue and NF-κB p65 and brain-derived neurotrophic factor (BDNF) in the hippocampal tissue. ResultsCompared with the blank group, the model group showed reductions in the time and frequency ratio of rats entering the elevated plus maze, the time and frequency of rats entering the central area of the open field, the time of entering the open box, the times of passing through the light and dark box, and the number of unburied beads (P<0.05, P<0.01). Compared with the model group, Fufang Kangjiaolv capsules ameliorated the anxiety of the model rats to varying degrees, and the high-dose group had the best effect, with increases in the proportions of time and frequency of rats entering the open arm in the elevated plus maze (P<0.05), the number of rats entering the central area in the open field (P<0.05), the time of entering the open box, the times of passing through the light and dark boxes, and the number of unburied beads (P<0.01). Moreover, the high-dose group showed alleviated pathological damage of hippocampal neurons and colon. The results of 16S rRNA gene sequencing showed that the model group had increased relative abundance of Firmicutes, Deferribacterota, Romboutsia, and Phascolarctobacterium, while it had decreased relative abundance of Bavcteroidota and Lactobacillus. The drug administration groups showed increased relative abundance of Bavcteroidota, Bacteroides, norank f norank o Clostridia UCG-014, and Blautia and decreased relative abundance of Firmicutes and Deferribacterota. Compared with the blank group, the model group showed down-regulated protein and mRNA levels of ZO-1 and occludin in the colon (P<0.01), elevated levels of TNF-α, IL-6, and IL-β in the colon, serum, and hippocampus (P<0.01), up-regulated protein level of NF-κB p65 in the colon and hippocampus (P<0.01), and down-regulated protein level of BDNF in the hippocampus (P<0.05). Compared with the model group, high-dose Fufang Kangjiaolv capsules up-regulated the mRNA levels of ZO-1 and occludin in the colon (P<0.01), lowered the levels of TNF-α, IL-6, and IL-β in the colon, serum, and hippocampus (P<0.01), up-regulated the protein levels of ZO-1 (P<0.01) and occludin (P<0.05) in the colon, down-regulated the protein level of NF-κB p65 in the colon and hippocampus (P<0.05), and up-regulated the protein level of BDNF in the hippocampus. ConclusionFufang Kangjiaolv capsules can reduce the anxiety-like behaviors in the rat model of CRS by regulating the gut microbiota disturbance, up-regulating the expression of tight junction proteins in the colon, repairing intestinal mucosal mechanical barrier, and down-regulating NF-κB/BDNF signaling pathway, thereby reducing peripheral and central inflammation. This study proves the hypothesis that Fufang Kangjiaolv capsules play an anti-anxiety role via the microbiota-gut-brain axis, providing a new idea for further research.


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