1.Comparative Study on Effect of Jingui Shenqiwan and Liuwei Dihuangwan on Reproductive Ability and Brain Function of Normal Mice
Hong SUN ; Fan LEI ; Chenggong LI ; Rui LUO ; Shixian HU ; Bin REN ; Juan HAO ; Yi DING ; Lijun DU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):1-14
ObjectiveTo explore the effects of Jingui Shenqiwan (JSW) and Liuwei Dihuangwan (LDW) on the reproductive ability and brain function of normal mice and compare the actions of the two medications. MethodsSeven groups of female and male mice were divided at a ratio of 2∶1. Except for the control group, the other six groups were as follows: a group of both males and females receiving JSW (3.0 g·kg-1), a group of both males and females receiving LDW (4.5 g·kg-1), a group of males receiving water and females receiving JSW, a group of males receiving water while females receiving LDW, a group of females receiving water while males receiving JSW, and a group of females receiving water while males receiving LDW. Each group was administered the drug for 14 days and then caged together at a 2∶1 (female∶male) ratio to detect the number of pregnant mice and calculate the pregnancy rate. Pregnant mice continued receiving the drug until they naturally gave birth, which was followed by the observation of newborn mice, calculation of their average number, and the measurement of the offspring's preference for sugar water and neonatal recognition index. At the end of the experiment, the weights of the thymus and spleen were measured to calculate the organ coefficients, and mRNA or protein expression was analyzed in the brain and testes or ovaries. A 1% sucrose solution was used to examine the euphoria of their brain reward systems, while novel object recognition test (NOR) was applied to assess their memory capabilities. mRNA expression was detected using real-time quantitative polymerase chain reaction (Real-time PCR) assay, and protein expression was analyzed with Western blot. ResultsCompared with the control group, oral administration of JSW to both male and female mice for 14 days significantly increased the pregnancy rate of female mice on day 2 after being caged together (P<0.05), while LDW showed a trend but no statistical significance. Additionally, compared with the control group, JSW could upregulate the gene expression of gonadotropin-releasing hormone (GnRH) in the thalamus, as well as reproductive stem cell factor (SCF) and tyrosine kinase receptor (c-Kit) in the testes and reproductive stem cell marker mouse vasa homologue (MVH) in the ovaries, upregulate the expression of proteins influencing neuronal functional activity, such as brain-derived neurotrophic factor (BDNF), in hippocampal neurons (P<0.05), and enhance sucrose preference in male mice (P<0.05). Compared with the control group, JSW significantly increased sucrose preference and novel object recognition index in offspring mice (P<0.05), which was related to the upregulation of hippocampal dopamine D1 receptor (D1R) and N-methyl-D-aspartate receptor (Nmdar) gene expression. Compared with the control group, both JSW and LDW could upregulate the protein expression of glucocorticoid receptor (GR), BDNF, and tyrosine kinase receptor B (TrkB) in the hippocampus of offspring mice (P<0.05). ConclusionJSW significantly enhances the reproductive ability of normal mice, which is not only related to the release of gonadotropin but also associated with its regulation of brain function. Additionally, JSW has a certain regulatory effect on the brain function of the offspring mice.
2.Quercetin Ameliorates Gouty Arthritis in Rats via ROS/NLRP3/IL-1β Signaling Pathway
Baowei FENG ; Yan WANG ; Chang LI ; Yujing ZHANG ; Dingxing FAN ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):145-153
ObjectiveTo investigate the effect of quercetin on acute gouty arthritis (GA) in rats by inhibiting the reactive oxygen species (ROS)/NOD-like receptor protein 3 (NLRP3)/interleukin-1β (IL-1β) signaling pathway. MethodsSixty SPF-grade male SD rats were randomized into normal, model, colchicine (0.3 mg·kg-1), and low-, medium-, and high-dose (25, 50, 100 mg·kg-1, respectively) quercetin groups (n=10). The rats in the dosing groups were administrated with the corresponding drugs (10 mL·kg-1) by gavage once a day for one week. An equal volume of normal saline was given by gavage to rats in normal and model groups. One hour after drug administration on day 5, an acute GA model was established in other groups except the control group via intra-articular injection of monosodium urate (MSU) suspension into the right posterior ankle joint cavity. The joint swelling and gait were scored at the time points of 6, 12, 24, 48 h after modeling. Histopathological alterations in the ankle joint tissue from each group were assessed by hematoxylin-eosin (HE) staining. Malondialdehyde (MDA), xanthine oxidase (XOD), and total superoxide dismutase (T-SOD) assay kits were used to assess the levels of MDA, XOD, and T-SOD in the serum. The levels of tumor interleukin-6 (IL-6), necrosis factor-α (TNF-α), and IL-1β in the rat serum, as well as ROS in the ankle joint tissue, were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was performed to determine the protein levels of NLRP3, thioredoxin-interacting protein (TXNIP), apoptosis-associated speck-like protein containing a CARD domain (ASC), precursor cysteinyl aspartate-specific proteinase-1 (pro-Caspase-1), cleaved Caspase-1 (Caspase-1 p20), and IL-1β in the ankle joint tissue. Real-time PCR was employed to assess the mRNA levels of TXNIP, NLRP3, ASC, IL-1β, and TNF-α in the ankle joint tissue. ResultsCompared with the normal group, the model group exhibited decreased spontaneous activity, mental fatigue, increased ankle joint swelling and gait scores (P<0.01), aggravated synovial tissue edema and inflammatory cell infiltration (P<0.01), elevated levels of XOD, MDA, TNF-α, IL-1β, and IL-6 in the serum and ROS in the joint tissue (P<0.01), a declined level of T-SOD (P<0.01), up-regulated protein levels of NLRP3, TXNIP, ASC, pro-Caspase-1, Caspase-1 p20, and IL-1β in the ankle joint tissue (P<0.01), and up-regulated mRNA levels of NLRP3, TXNIP, ASC, IL-1β, and TNF-α in the ankle joint tissue (P<0.01). Compared with the model group, the medium- and high-dose quercetin groups showed improved general conditions, decreased gait scores (P<0.05, P<0.01), reduced joint swelling (P<0.01), alleviated synovial tissue edema and inflammatory cell infiltration (P<0.05, P<0.01), lowered levels of XOD, MDA, TNF-α, IL-1β, and IL-6 in the serum and ROS in the joint tissue (P<0.01), increased levels of T-SOD (P<0.01), down-regulated protein levels of TXNIP, NLRP3, ASC, pro-Caspase-1, Caspase-1 p20, and IL-1β in the ankle joint tissue (P<0.05, P<0.01), and down-regulated mRNA levels of TXNIP, NLRP3, ASC, IL-1β, and TNF-α in the ankle joint tissue (P<0.01). Low-dose quercetin also ameliorated some of the above parameters (P<0.05, P<0.01). ConclusionQuercetin exerts anti-GA effects by blocking the ROS/NLRP3/IL-1β signaling pathway, downregulating NLRP3 inflammasome activation, and inhibiting the production of pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6.
3.Sanren Runchang Formula Regulates Brain-gut Axis to Treat IBS-C: A Randomized Controlled Trial
Teng LI ; Xinrong FAN ; He YAN ; Zhuozhi GONG ; Mengxi YAO ; Na YANG ; Yuhan WANG ; Huikai HU ; Wei WEI ; Tao LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):154-161
ObjectiveTo observe the clinical efficacy of Sanren Runchang formula in treating constipation-predominant irritable bowel syndrome (IBS-C) by regulating the brain-gut axis and the effects of the formula on serum levels of 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP), and substance P (SP). MethodsA randomized controlled design was adopted, and 72 IBS-C patients meeting Rome Ⅳ criteria were randomized into observation and control groups (36 cases).The observation group received Sanren Runchang formula granules twice daily, and the control group received lactulose oral solution daily for 4 weeks. IBS Symptom Severity Scale (IBS-SSS), IBS Quality of Life Scale (IBS-QOL), and Bristol Stool Form Scale (BSFS) were used to assess clinical symptoms, and bowel movement frequency was recorded. The Self-Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS) were employed to evaluate psychological status. ELISA was employed to measure the serum levels of 5-HT, VIP, and SP. ResultsThe total response rate in the observation group was 91.67% (33/36), which was higher than that (77.78%, 28/36) in the control group (χ2=4.50, P<0.05). After treatment, both groups showed increased defecation frequency and BSFS scores, decreased IBS-SSS total score, abdominal pain and bloating scores, IBS-QOL health anxiety, anxiety, food avoidance, and behavioral disorders scores, SAS and SDS scores, serum 5-HT and VIP levels, and increased SP levels (P<0.05, P<0.01). Moreover, the observation group showed more significant changes in the indicators above than the control group (P<0.05, P<0.01). The SP level showed no significant difference between the two groups. During the 4-week follow-up, the recurrence rate was 5.88% in the observation group and 31.25% in the control group. No adverse events occurred in observation group, and 2 cases of mild diarrhea occurred in the control group. ConclusionSanren Runchang formula demonstrated definitive efficacy in alleviating gastrointestinal symptoms and improving the psychological status and quality of life in IBS-C patients, with a low recurrence rate. The formula can regulate serum levels of neurotransmitters such as 5-HT and VIP, suggesting its potential regulatory effect on the brain-gut axis through modulating neurotransmitters and neuropeptides. However, its complete mechanism of action requires further investigation through detection of additional brain-gut axis-related biomarkers.
4.Analysis of the application status of prescription pre-review systems in Yunnan province
Fan XU ; Wenjie YIN ; Kejia LI ; Zhengfu LI ; Jie CHEN ; Meixian WU ; Ruixiang CHEN ; Songmei LI ; Guowen ZHANG ; Te LI
China Pharmacy 2026;37(1):6-10
OBJECTIVE To investigate the application status of prescription pre-review systems in healthcare institutions of Yunnan province, evaluate their system functions and management capabilities, and provide a practical basis for promoting rational drug use. METHODS A questionnaire survey was conducted among public healthcare institutions at or above the secondary level in Yunnan province to investigate the deployment status of the systems. A capability maturity assessment framework was constructed, encompassing 6 dimensions and 39 indicators, including real-time prescription review, prescription correlation review, rule setting, evidence-based information support, prescription authority management, and system operation management. This framework was then used to evaluate the institutions that had implemented the pre-review systems. RESULTS A total of 100 valid questionnaires were collected, with 37 institutions having adopted prescription pre-review systems, mainly tertiary hospitals. The system predominantly adopted a modular architecture and was embedded into the hospital information system through application programming interfaces and middleware, providing certain capabilities for real-time prescription risk identification. Evaluation results indicated that basic functions such as reviewing indications, contraindications, and drug compatibility performed well, while deficiencies remained in functions related to parenteral nutrition prescription, review of drug dosage for specific diseases, individual patient characteristic recognition, and rule setting. Moreover, the construction of review centers and establishment of management systems were also not well-developed. CONCLUSIONS The overall application rate of prescription pre-review systems in Yunnan province remains low. System functions and management mechanisms require further improvement. It is recommended to enhance information infrastructure in lower-level institutions and explore regionally unified review models to promote standardized and intelligent development of prescription review practices.
5.Intervention Effects of Formulas Containing Ephedrae Herba and Armeniacae Semen Amarum on Asthma in Mice and Their Regulatory Effects on Thermosensitive TRP Channels
Mengwen LI ; Yuhao FAN ; Xinsheng FAN ; Yaojing YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):32-40
ObjectiveTo investigate the differences in the regulatory effects of formulas containing Ephedrae Herba and Armeniacae Semen Amarum (Mahuangtang, Sanaotang, and Maxing Shigantang) on thermosensitive transient receptor potential ion channels (thermo TRPs) in the mouse model of asthmatic airway inflammation. MethodsSixty female C57BL/6 mice were allocated into blank, model, dexamethasone (0.75 mg·kg-1), Mahuangtang (3.8 g·kg-1), Sanaotang (2.8 g·kg-1), and Maxing Shigantang (6.6 g·kg-1) groups (n=10). The mouse model of asthma was established with ovalbumin (OVA) and treated with normal saline (blank group) or corresponding drugs (10 mL·kg-1), once a day, 19-28 days after modeling. The levels of eosinophils (EOS) in peripheral blood and white blood cell (WBC) in bronchoalveolar lavage fluid (BALF), changes in enhanced pause (Penh), and pathological damage of lung tissue were observed in each group. Western blot and real-time PCR were employed to quantify the protein and mRNA levels, respectively, of high-temperature thermosensitive channels (TRPV1 and TRPV3) and low-temperature thermosensitive channels (TRPA1 and TRPM8) in the lung tissue. ResultsCompared with the blank group, the model group showed a typical asthma phenotype, including elevations in the level of EOS in peripheral blood, level of WBC in BALF, and value of Penh (P<0.05,P<0.01), and severe lung tissue damage. Compared with the model group, the three formulas alleviated the asthma phenotype to varying degrees (P<0.05,P<0.01). Compared with the blank group, the model group showed up-regulated protein levels of TRPV1 and TRPA1 in the lung tissue (P<0.01). Compared with the model group, Maxing Shigantang and Sanaotang groups showed down-regulated protein levels of TRPV1 and TRPA1 (P<0.05, P<0.01). Moreover, Maxing Shigantang and Sanaotang groups showed more significant down-regulation in protein levels of TRPV1 and TRPA1, respectively (P<0.01), while no obvious regulatory effect was observed in the Mahuangtang group. Compared with those in the blank group, the protein levels of TRPV3 and TRPM8 were up-regulated in the model group (P<0.01). Compared with the model group, Maxing Shigantang and Sanaotang down-regulated the protein levels of TRPV3 and TRPM8 (P<0.01). Moreover, Maxing Shigantang and Sanaotang exerted stronger down-regulating effects on TRPV3 (P<0.05) and TRPM8 (P<0.01), respectively. Compared with the blank group, the model group presented up-regulated mRNA levels of TRPV1, TRPV3, TRPA1, and TRPM8 in the lung tissue (P<0.01), and such up-regulations were significantly decreased by Maxing Shigantang and Sanaotang (P<0.01). Moreover, Maxing Shigantang outperformed Sanaotang in regulating high-temperature thermosensitive channels TRPV1 and TRPV3 (P<0.05, P<0.01). The regulation effect of the, Maxing Shigantang on high-temperature thermosensory channel proteins of TRPV1 and TRPV3 was better than that of the Sanaotang P<0.05P<0.01while the Sanaotang outperformedhad a significant regulatory effect on Maxing Shigantang in regulating the low-temperature thermosensory thermosensitive channel proteins of TRPA1 and TRPM8which was better than that of the Maxing Shigantang (P<0.05,P<0.01). ConclusionThe experimental results showed that Mahuangtang, Sanaotang, and Maxing Shigantang all had protective effects on asthma airway inflammation.while Mahuangtang did not show the regulatory effect on TRPV1 and or TRPA1. Maxing Shigantang preferred to regulate high-temperature thermosensory thermosensitive channels of TRPV1 and TRPV3 channels, and Sanaotang preferred to regulate low-temperature thermosensory thermosensitive channels of TRPA1 and TRPM8.
6.Exploration of Traditional Chinese Medicine Syndrome Characteristics in A Heart Failure Model Induced by Coronary Artery Ligation Based on Method of Syndrome Identification by Prescription Efficacy
Xiaoqian LIAO ; Peiyao LI ; Xingyu FAN ; Zhenyu ZHAO ; Junyu ZHANG ; Yuehang XU ; Zhixi HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):169-177
Chronic heart failure (CHF) is a major global public health problem, and myocardial infarction is one of its main causes. The mouse model of heart failure induced by coronary artery ligation is widely used in the study of CHF, while the TCM syndrome attributes of this model have not yet been clarified. According to the theory of correspondence between prescriptions and syndromes, the method of syndrome identification by prescription efficacy is an important means of current syndrome research of animal models. This method deduces the syndrome characteristics of animal models through prescription efficacy. Taking the four basic syndrome elements of Qi, blood, Yin and Yang as the classification reference, this study used coronary artery ligation to construct a mouse model of CHF and treated the model with four representative TCM injections with the effects of replenishing Qi, warming Yang, nourishing Yin, and activating blood and enalapril. Echocardiography, tongue color parameters, histopathology, serum N-terminal pro-brain natriuretic peptide (NT-proBNP) and cardiac troponin Ⅰ (cTnⅠ) levels, and systematically explored the TCM syndrome attributes of this model. The results showed that the coronary ligation model presented an obvious cardiac function decline, myocardial fibrosis, infarct size expansion, and purple dark tongue, which were consistent with the basic syndrome characteristics of blood stasis in CHF. Danhong injection had significant effects of improving the cardiac function, alleviating myocardial fibrosis, and reducing serum NT-proBNP and cTnⅠ levels. Huangqi Injection and Shenfu injection can improve the cardiac function and tongue color parameters, with limited effects. The effect of Shenmai injection group was not obvious. This study verifies that the established model conforms to blood stasis syndrome through the method of syndrome identification by prescription efficacy, which provides an experimental basis for the study of TCM syndrome mechanism of CHF.
7.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
8.Overview of Diagnosis,Treatment and Mechanism Research of Functional Dyspepsia by Integrated Traditional Chinese and Western Medicine
Shengsheng ZHANG ; Zhaohong SHI ; Xiaofang LU ; Luqing ZHAO ; Danyan LI ; Shu ZHANG ; Lu ZHAO ; Yudi ZHUO ; Nian WANG ; Fan LIU ; Shuangyi LI ; Xudong TANG
Journal of Traditional Chinese Medicine 2026;67(4):397-403
Functional dyspepsia (FD) is a prioritized disease category where traditional Chinese medicine (TCM) demonstrates distinct therapeutic advantages. The current western medicine treatment for FD is mainly based on proton pump inhibitors and prokinetic agents, with digestive enzymes, probiotics and antidepressants serving as adjuvant medication, yet such therapies still have certain limitations. TCM treatment for FD includes oral administration of Chinese herbal formulas and Chinese patent medicines, as well as external TCM therapies such as acupuncture and moxibustion, acupoint application, hot medicinal compress therapy, rubbing with ointment, medicinal iontophoresis, auricular acupoint therapy and tui na (Chinese medical massage). The combined treatment of FD with integrated TCM and western medicine can significantly improve clinical effectiveness and reduce adverse reactions. The common mechanisms underlying the therapeutic effects of both TCM and western medicine revolve around the core pathological processes of FD, mainly focusing on restoring gastrointestinal motility, regulating the levels of brain-gut peptides, modulating intestinal microecology, and ameliorating inflammatory status. The differential mechanisms lie in the precise targeting feature of western medicine versus the holistic-regulating and multi-target characteristics of TCM, and the two approaches exert a synergistic effect to enhance efficacy. This paper proposes to leverage the advantages of TCM in holistic regulation and the strengths of western medicine in targeted treatment, so as to provide personalized and comprehensive treatment regimens for FD patients.
9.Experience Discussion on Feeding, Breeding and Genetic Management of Common Marmosets (Callithrix jacchus)
Li LI ; Xuebo LI ; Yongheng FAN ; Donghua HE ; Jianhong LI ; Liangtang CHANG
Laboratory Animal and Comparative Medicine 2026;46(1):107-118
The common marmoset (Callithrix jacchus) is a species of the genus Callithrix in the primate family Callitrichidae. Common marmosets are an important scientific research resource for the study of neuroscience, medicine, and pharmacy. The feeding and breeding of common marmosets provide important support for scientific research. At present, common marmoset resources in China are at a critical stage of development. This paper takes the Center for Excellence in Brain Science and Intelligence Technology of the Chinese Academy of Sciences (hereafter referred to as CEBSIT) as an example to discuss the experience of common marmoset feeding, breeding and genetic management. In terms of breeding environment and facilities, on the basis of the national standard, more stringent environmental parameter standards are implemented for environmental temperature, relative humidity, the number of air exchanges, and the average concentration of sedimentation bacteria. Two kinds of cages for common marmoset breeding and experimentation are designed. According to the differences in facility conditions and staffing, a cleaning and maintenance method combining "dry rearing" and "wet rearing" is introduced. In terms of feeding management, in order to meet the high nutritional needs of common marmosets, CEBSIT prepares special formulated diets, and introduces the transition method of common marmosets from diets mixed with formulated rations and pellets to exclusive commercial diets, and strictly regulates the operation procedures of formulated diets for common marmosets, and has accumulated a series of methods for preventing nutritional and metabolic diseases in common marmosets. In terms of animal health management, the health of common marmosets is ensured through measures such as regular quarantine, isolation and treatment, and epidemic prevention, and the health inspection process of common marmosets is described in detail, and hierarchical and classified management is carried out according to the physical condition of common marmosets. This paper also briefly describes the treatment of diarrhea and pneumonia that are common in common marmosets. In terms of breeding management, it covers the selection of breeding common marmosets, combining pairs of common marmosets into the same cage for breeding, group splitting and hand-rearing of infant common marmosets, and introduces the method of reintroducing hand-reared infant common marmosets to parental care from the perspective of animal welfare. In terms of genetic management, the common marmoset population in CEBSIT is maintained as a closed colony and follows a non-inbreeding method. CEBSIT has developed an inbreeding screening system to check pedigree information before pairing to avoid inbreeding. Under conditions without introducing new individuals from outside, CEBSIT has continuously bred the colony for 5 generations while maintaining genetic heterozygosity. This paper only discusses the feeding, breeding and genetic management of the common marmoset and aims to provide a useful reference for other institutions engaged in common marmoset breeding.
10.Curcumin-loaded exosomes from hypoxia-treated mesenchymal stem cells alleviate microglial inflammatory response in a combined therapy approach
Xiaobin HUANG ; Qianqian LI ; Peng ZHANG ; Yanhua ZHOU ; Anran FAN
Acta Universitatis Medicinalis Anhui 2026;61(1):104-112
ObjectiveTo investigate the effects of hypoxia-treated mesenchymal stem cell (MSCs) exosomes (Exo) and their loading with curcumin on microglial inflammatory responses, and to explore the enhancing effect of hypoxia treatment on the function of MSCs Exo. MethodsThe supernatants of human umbilical cord (hUC)-MSCs cultured under normal and hypoxic conditions were collected, and Exo were isolated using ultracentrifugation. After identification by transmission electron microscopy and Western blot, curcumin was loaded using the co-incubation method. The lipopolysaccharide (LPS)-induced microglial inflammation model was treated with dimethyl sulfoxide (DMSO), curcumin, normoxia Exo, hypoxia Exo, normoxic Exo loaded with curcumin, and hypoxic Exo loaded with curcumin, respectively. The expression of the M1-type marker inducible nitric oxide synthase (iNOS) in BV2 cells was detected by immunofluorescence (IF). Western blot and enzyme-linked immunosorbent assay (ELISA) were used to measure the expression and secretion levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 in the cells and their culture supernatants. ResultsNormoxia Exo, hypoxia Exo, normoxic Exo loaded with curcumin, and hypoxic Exo loaded with curcumin exhibited a "saucer-like" shape with a diameter ranging from 30~150 nm, and the expression of exosomal markers CD9, CD81, and TSG101 were positive. After treating the BV2 cell inflammation model, IF results showed that, compared with the normoxia Exo group, treatment with hypoxic Exo significantly reduced the expression of iNOS. Moreover, when compared with the curcumin group and the normoxic Exo loaded with curcumin group, the expression level of iNOS significantly decreased after treatment with hypoxic Exo loaded with curcumin. The results of Western blot and ELISA indicated that, in comparison with the normoxia Exo group, treatment with hypoxic Exo significantly reduced the expression and secretion of the inflammatory cytokines TNF-α, IL-1β, and IL-6. Additionally, when compared with the curcumin group and the normoxic Exo loaded with curcumin group, both the expression and secretion of TNF-α, IL-1β, and IL-6 significantly decreased after treatment with hypoxic Exo loaded with curcumin. ConclusionHypoxia preconditioning can enhance the ability of hUC-MSCs-Exo in the inhibition of microglial polarization and inflammatory factors’ secretion. Additionally, using Hypoxia-MSCs-Exo as a drug-delivery carrier of curcumin can improve its solubility and stability, facilitating its absorption by cells and exerting the therapeutic effect of combination therapy.

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