1.Mechanism of Yangjing Zhongyutang in Regulating SIRT1/PGC-1α Signaling Pathway to Promote Mitochondrial Function and Alleviate Oxidative Stress Damage in Rats with Diminished Ovarian Reserve
Ping ZHANG ; Lijuan YANG ; Shenghui CHEN ; Wenliang YAO ; Yuliang ZHOU ; Ling MA ; Huiying WU ; Yanwen XU ; Ziyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):46-55
ObjectiveTo observe the effects of Yangjing Zhongyutang (YJZYT) on mitochondrial biogenesis and oxidative stress damage mediated by the silent information regulator 1 (SIRT1)/peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α) signaling pathway in cyclophosphamide (CTX)-induced rats with diminished ovarian reserve (DOR), and to explore its mechanism in improving ovarian reserve function and follicular development. MethodsForty-two 8-week-old female SD rats with normal estrous cycles were randomly divided into a blank control group (n=7) and a model group (n=35). Rats in the model group received a single intraperitoneal injection of CTX (90 mg·kg-1) to establish the DOR model. After modeling, estrous cycles were monitored for 7 consecutive days, and model success was confirmed based on criteria for estrous cycle disruption. After successful modeling, rats were divided into groups for intervention: estradiol valerate group (0.09 mg·kg-1), and YJZYT high-, medium-, and low-dose groups (19.98, 9.99, 5.00 g·kg-1). The blank control group and model group were given an equal volume of distilled water by gavage. All groups received daily gavage once for 4 consecutive weeks. The general state, body weight, and ovarian wet weight of rats were observed and recorded, and the ovarian organ index was calculated. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Hematoxylin-eosin (HE) staining was performed to observe ovarian histomorphological changes and follicular development status. Immunofluorescence was used to detect reactive oxygen species (ROS) expression levels. Colorimetric assays were employed to measure adenosine triphosphate (ATP) and malondialdehyde (MDA) content in ovarian tissues. Quantitative Real-time polymerase chain reaction (Real-time PCR) was used to detect mitochondrial DNA (mtDNA) copy number and the mRNA expression levels of key genes including SIRT1, PGC-1α, nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM). Western blot was performed to detect the protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM. ResultsCompared with the blank group, rats in the model group exhibited disrupted estrous cycles, obviously reduced body weight, and decreased ovarian index (P<0.05). Ovarian histopathology revealed cortical thinning, loose structure, and a significant reduction in both primordial and growing follicles (P<0.01). Serum FSH and LH levels were significantly elevated (P<0.01), while E2 and AMH levels were obviously reduced (P<0.05, P<0.01). ATP content and mtDNA copy number decreased in ovarian tissue (P<0.01), ROS expression increased, MDA levels rose, while SOD and GSH-Px activities obviously decreased (P<0.05, P<0.01), mRNA and protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM were obviously downregulated (P<0.05, P<0.01). After treatment, compared with the model group, body weight and ovarian index obviously recovered in rats administered various doses of YJZYT (P<0.05), serum E2 and AMH levels increased, while FSH and LH levels obviously decreased (P<0.05, P<0.01), ovarian tissue ATP content and mtDNA copy number were up-regulated, ROS and MDA levels decreased, and antioxidant enzymes SOD and GSH-Px activity obviously increased (P<0.05, P<0.01), Gene and protein expression levels related to the SIRT1/PGC-1α /NRF1/TFAM signaling pathway were obviously up-regulated compared to the model group (P<0.05, P<0.01), HE staining revealed that ovarian structure gradually recovered to integrity in all treatment groups, with a obviously increase in the number of primordial and growing follicles (P<0.05, P<0.01). Granulosa cells were neatly arranged, indicating marked improvement in ovarian function. ConclusionYJZYT may improve ovarian function and follicular development in rats with diminished ovarian reserve by activating the SIRT1/PGC-1α signaling pathway, promoting mitochondrial biogenesis, enhancing mitochondrial function, and alleviating oxidative stress damage.
2.Association of vegetables and fruits intake with depressive symptoms in Yunnan multi ethnic first year junior high school students
CHEN Yiyao*, DU Yeming, YIN Wan, HUANG Jianhong, LIU Zihan, JIANG Yinghong, YU Qiaoying, PAN Lijuan, XU Honglü ;
Chinese Journal of School Health 2026;47(1):32-36
Objective:
To explore the association of vegetables and fruits intake with depressive symptoms among multi ethnic first year junior high school students in Yunnan Province, so as to provide data support for preventing and reducing depressive symptoms among first year junior high school students.
Methods:
From October to December 2022, a cluster random sampling method was used to select 8 500 first year junior high school students from 11 ethnic minority areas in Yunnan Province (Fugong County, Longling County, Longyang District, Luchun County, Mojiang County, Nanjian County, Qiaojia County, Shuangjiang County, Tengchong City, Yuanmou County, Zhenyuan County), to investigate with a questionnaire. The Dietary Frequency Questionnaire was used to collect dietary behavior datas, and the Chinese version of Depression Anxiety Stress Scale-21 (DASS-21) was used to assess depressive symptoms. The generalized linear model was used to analyze the association of vegetable and fruit intake with depressive symptoms in students, and stratified analysis was performed according to ethnicity.
Results:
The detection rate of depressive symptoms among first year junior high school students in Yunnan Province was 29.5%. The detection rates of depressive symptoms in Han and minority first year junior high school students were 26.9% and 31.6%. After controlling for demographic variables such as gender, age, family residence and other confounding factors, the generalized linear model analysis results showed that the intake of leafy vegetables ( β= -0.07 , 95%CI =-0.12 to -0.01), flat fruits ( β=-0.06, 95%CI =-0.12 to -0.00) and hot natured fruits ( β=0.11, 95%CI = 0.04- 0.17) were associated with depressive symptoms in Han first year junior high school students (all P <0.05). The intake of melon and fruit vegetables ( β=-0.06, 95%CI =-0.11 to -0.01) and hot natured fruits ( β=0.06, 95%CI =0.01-0.12) were associated with depressive symptoms in ethnic minority first year junior high school students (both P <0.05).
Conclusions
The intake of vegetables and fruits among multi ethnic first year junior high school students in Yunnan Province is related to the risk of depressive symptoms. It is suggested to strengthen the consumption guidance and education of vegetables and fruits to prevent depressive symptoms among first year junior high school students.
3.Association of personality and sleep quality with psychological distress of junior and senior high school stduents
Chinese Journal of School Health 2026;47(1):65-69
Objective:
To explore the effects of personality and sleep quality with psychological distress of junior and senior high school stduents, so as to provide a reference basis for precise interventions of junior and senior high school students mental health.
Methods:
In October 2023, a convenience sampling method was used to select 9 034 students aged 12-17 from Shiyan City as the study subjects. The Pittsburgh Sleep Quality Index (PSQI) and Kessler Psychological Distress Scale (K10) were used to collect information on sleep quality and psychological distress of junior and senior high school stduents. Between group comparison was conducted by using t-test and Chi-square test. Generalized linear models were employed to analyze the interaction and joint effects of personality and sleep quality on psychological distress.
Results:
The generalized linear model analysis showed that the interaction between personality and sleep quality on psychological distress was statistically significant of junior and senior high school students(effect size=0.80, P <0.01). The general linear model analysis indicated that, after adjusting for variables such as age, gender, screen time, and daily sitting time with the extroverted and good sleep quality group as the reference, the introverted and poor sleep quality group had the largest mean difference in psychological distress scores (difference=0.51, P <0.05). When stratified by sleep quality, psychological distress scores were higher in the introverted and neutral personality groups with both poor and good sleep quality compared to the extroverted group (poor sleep quality: introverted difference=3.71, neutral difference=1.14; good sleep quality: introverted difference=2.23, neutral difference=0.57, all P < 0.05). When stratified by personality, psychological distress scores were higher in the poor sleep quality groups for introverted, neutral, and extroverted individuals compared to their good sleep quality counterparts (differences=8.66, 7.83, 7.34, all P < 0.05 ).
Conclusions
Personality and sleep quality have interactive and joint effects on psychological distress of junior and senior high school stduents. Personalized psychological interventions should be developed based on personality and sleep quality.
4.Components and Brain-protective Effect of Chuanxiong Rhizoma-Paeoniae Radix Rubra in Improving Ischemic Stroke Based on UPLC-Q-TOF-MS
Qizhong JIN ; Jie ZHANG ; Lijuan XIU ; Fan XU ; Lei WANG ; Ning WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):20-29
ObjectiveTo investigate the chemical constituents of Chuanxiong Rhizoma-Paeoniae Radix Rubra(CRPRR) that cross the blood-brain barrier in rats with ischemic stroke, their brain-protective effects, and their impact on inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and pharmacodynamic experiments. MethodsA focal cerebral ischemia-reperfusion injury model was established in rats via the middle cerebral artery occlusion/reperfusion (MCAO/R) method using intraluminal suture. Neurological function was evaluated using behavioral scoring. UPLC-Q-TOF-MS was employed to identify the chemical constituents of CRPRR that crossed the blood-brain barrier and entered the cerebrospinal fluid in MCAO/R model rats. Male Sprague-Dawley rats were randomly divided into six groups: sham operation group, model group, low-, medium-, and high-dose CRPRR groups (1.35, 2.7, 5.4 g·kg-1, respectively), and an edaravone group (5 mg·kg-1), with 12 rats in each group. The sham and model groups received normal saline, while the treatment groups received the respective doses of CRPRR once daily by gavage for three consecutive weeks. The brain-protective effects of CRPRR were assessed using the Longa five-point scoring method, open field test, Morris water maze, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin (HE) staining, and transmission electron microscopy. ResultsNine chemical constituents were identified in the cerebrospinal fluid containing CRPRR, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide. Animal experiment results showed that compared with the sham operation group, the model group exhibited disordered neuronal arrangement, severe vacuolation, nuclear pyknosis, and evident mitochondrial swelling. Chromatin aggregation and peripheralization were also observed. Neurological scores and the number of crossings in the central region were significantly increased (P<0.01), while platform crossings were significantly decreased (P<0.01), and clear infarct areas were present (P<0.01). Serum levels and protein expression of TNF-α, IL-1β, and IL-18 were significantly elevated (P<0.01). Compared with the model group, all dose groups of CRPRR showed marked improvement in neuronal morphology which was close to the normal level, with mitochondrial swelling alleviated and chromatin distribution more uniform. The medium- and high-dose groups significantly reduced neurological scores (P<0.01), while the low-, medium-, and high-dose groups significantly reduced the number of central crossings (P<0.01) and infarct volume (P<0.01), and decreased TNF-α, IL-1β, and IL-18 levels (P<0.05, P<0.01) compared with the model group. Furthermore, the medium- and high-dose groups significantly reduced TNF-α protein expression (P<0.05,P<0.01), and the high-dose group significantly reduced IL-1β and IL-18 protein expression (P<0.01). ConclusionThis study confirmed that CRPRR improves neurological function and alleviates brain tissue damage in MCAO/R rats. Its mechanism may be associated with the downregulation of inflammatory factors TNF-α, IL-1β, and IL-18, as well as the presence of nine active chemical constituents in cerebrospinal fluid, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide, which are closely related to their brain-protective effects.
5.Components and Brain-protective Effect of Chuanxiong Rhizoma-Paeoniae Radix Rubra in Improving Ischemic Stroke Based on UPLC-Q-TOF-MS
Qizhong JIN ; Jie ZHANG ; Lijuan XIU ; Fan XU ; Lei WANG ; Ning WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):20-29
ObjectiveTo investigate the chemical constituents of Chuanxiong Rhizoma-Paeoniae Radix Rubra(CRPRR) that cross the blood-brain barrier in rats with ischemic stroke, their brain-protective effects, and their impact on inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and pharmacodynamic experiments. MethodsA focal cerebral ischemia-reperfusion injury model was established in rats via the middle cerebral artery occlusion/reperfusion (MCAO/R) method using intraluminal suture. Neurological function was evaluated using behavioral scoring. UPLC-Q-TOF-MS was employed to identify the chemical constituents of CRPRR that crossed the blood-brain barrier and entered the cerebrospinal fluid in MCAO/R model rats. Male Sprague-Dawley rats were randomly divided into six groups: sham operation group, model group, low-, medium-, and high-dose CRPRR groups (1.35, 2.7, 5.4 g·kg-1, respectively), and an edaravone group (5 mg·kg-1), with 12 rats in each group. The sham and model groups received normal saline, while the treatment groups received the respective doses of CRPRR once daily by gavage for three consecutive weeks. The brain-protective effects of CRPRR were assessed using the Longa five-point scoring method, open field test, Morris water maze, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin (HE) staining, and transmission electron microscopy. ResultsNine chemical constituents were identified in the cerebrospinal fluid containing CRPRR, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide. Animal experiment results showed that compared with the sham operation group, the model group exhibited disordered neuronal arrangement, severe vacuolation, nuclear pyknosis, and evident mitochondrial swelling. Chromatin aggregation and peripheralization were also observed. Neurological scores and the number of crossings in the central region were significantly increased (P<0.01), while platform crossings were significantly decreased (P<0.01), and clear infarct areas were present (P<0.01). Serum levels and protein expression of TNF-α, IL-1β, and IL-18 were significantly elevated (P<0.01). Compared with the model group, all dose groups of CRPRR showed marked improvement in neuronal morphology which was close to the normal level, with mitochondrial swelling alleviated and chromatin distribution more uniform. The medium- and high-dose groups significantly reduced neurological scores (P<0.01), while the low-, medium-, and high-dose groups significantly reduced the number of central crossings (P<0.01) and infarct volume (P<0.01), and decreased TNF-α, IL-1β, and IL-18 levels (P<0.05, P<0.01) compared with the model group. Furthermore, the medium- and high-dose groups significantly reduced TNF-α protein expression (P<0.05,P<0.01), and the high-dose group significantly reduced IL-1β and IL-18 protein expression (P<0.01). ConclusionThis study confirmed that CRPRR improves neurological function and alleviates brain tissue damage in MCAO/R rats. Its mechanism may be associated with the downregulation of inflammatory factors TNF-α, IL-1β, and IL-18, as well as the presence of nine active chemical constituents in cerebrospinal fluid, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide, which are closely related to their brain-protective effects.
6.Study on determination of multi-ingredient contents in Yinchen Wuling Powder based on HPLC fingerprinting and chemical pattern recognition techniques
Lijuan ZHAO ; Yanyun WEI ; Liang GUO ; Xiaoyan XU ; Xinbo PAN
International Journal of Traditional Chinese Medicine 2025;47(2):221-228
Objective:To establish the HPLC fingerprints and multi-component content determination method of Yinchen Wuling Powder; To evaluate the quality of Yinchen Wuling Powder combined with chemical pattern recognition technology.Methods:A wavelength-switching HPLC method was used to establish the fingerprint profiles of different batches of Yinchen Wuling Powder and determine the contents of seven components of neochlorogenic acid, chlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, cinnamic acid, cinnamaldehyde. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were applied to evaluate the quality. Quality difference biomarkers were screened.Results:The fingerprint profiles of 24 batches of Yinchen Wuling Powder were identified with 20 common peaks, and 7 components were recognized. The similarity between the 24 batches of samples and the control profile was 0.966-0.994, and the attribution analysis showed that peaks 1, 2, 3 were common to Artemisia Scoparia and Alismatis Rhizoma. Chromatographic peaks 4 (neochlorogenic acid), 5, 6, 7 (chlorogenic acid), 8, 9, 10, 11, 12, 13, 14 (isochlorogenic acid B), 15 (isochlorogenic acid A), 16 (isochlorogenic acid C), and 19 were from Artemisia Scoparia, and peaks 17, 18 and 20 were from Cinnamomi Ramulus. Principal component analysis classified the 24 batches of samples into 4 categories, and partial least squares discriminant analysis was used to determine 11 quality difference biomarkers, including isochlorogenic acid B and neochlorogenic acid.Conclusions:The established method of determining fingerprint and multi-component content is accurate and with good repeatability. The results can provide a certain basis for the quality evaluation of Yinchen Wuling Powder.
7.Effect of Auricular Point Seed-Pressing and Earpoint Scraping on Sleep Quality,Sleep Structure Index and Neurotransmitter in Patients with Sleep Disorders in Neurology Department
Yue BAO ; Bingguo XU ; Hui ZHANG ; Lijuan WANG ; Guangyue PENG ; Mengmeng GU ; Huidi XU
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(6):1434-1440
Objective To investigate the effects of auricular point seed-pressing and earpoint scraping on sleep quality,sleep structure index and neurotransmitter in patients with sleep disorders in neurology department.Methods A total of 110 patients with sleep disorders treated in the Department of Neurology at Nanjing Hospital Affiliated to Nanjing Medical University from June 2022 to June 2024 were selected as the study subjects.The patients were randomly divided into an observation group and a control group using a random number table,with 55 cases in each group.The control group was treated with Estazolam Tablets,while the observation group received auricular point seed-pressing therapy combined with ear scraping therapy.The treatment course lasted for 4 weeks.After treatment,the clinical efficacy of the two groups was evaluated.Changes in the Pittsburgh Sleep Quality Index(PSQI)scores were observed before and after treatment,as well as sleep structure indicators,including total sleep time(TST),sleep latency(SL),arousal index(AI),non-rapid eye movement sleep time(NREM)[light sleep stage(S1),light to moderate sleep stage(S2),moderate sleep stage(S3),deep sleep stage(S4)],actual sleep time(AST),sleep efficiency(SE),and rapid eye movement sleep time(REM).The serum levels of norepinephrine(NE),5-hydroxytryptamine(5-HT),and gamma-aminobutyric acid(GABA)were compared before and after treatment,as well as changes in the Quality of Life Scale(SF-36)scores.The safety and incidence of adverse reactions in the two groups were also evaluated.Results(1)The total effective rate in the observation group was 96.61%(53/55),while it was 81.82%(45/55)in the control group.The efficacy of the observation group was superior to that of the control group,with a statistically significant difference(P<0.05).(2)After treatment,the PSQI scores of both groups were significantly improved(P<0.05),and the observation group showed significantly better improvement in PSQI scores compared to the control group,with a statistically significant difference(P<0.05).(3)After treatment,the sleep structure indicators in the observation group,including TST,SL,AI,S1,S2,S3,S4,AST,SE,and REM,were significantly improved,and the differences were statistically significant compared to the control group(P<0.05).(4)After treatment,the serum levels of NE,5-HT,and GABA in both groups were significantly improved,and the observation group showed significantly better improvement in these levels compared to the control group,with a statistically significant difference(P<0.05).(5)After treatment,the SF-36 scores of both groups were significantly improved,and the observation group showed significantly better improvement in SF-36 scores compared to the control group,with a statistically significant difference(P<0.05).(6)The total incidence of adverse reactions in the observation group was 0.00%(0/55),while it was 10.91%(6/55)in the control group.The incidence of adverse reactions in the observation group was significantly lower than that in the control group,with a statistically significant difference(P<0.05).Conclusion Auricular point seed-pressing therapy combined with ear scraping therapy for the treatment of sleep disorders at the Department of Neurology can significantly improve patients'sleep quality,enhance sleep structure-related indicators,regulate neurotransmitter levels,thereby improving patients'quality of life with high safety and efficacy.
8.The research progress on the improvement effect of repeated peripheral magnetic stimulation on upper limb and lower limb spasm after stroke.
Lijuan XU ; Liang YE ; Jie JIA ; Shugeng CHEN
Journal of Biomedical Engineering 2025;42(3):628-634
Post-stroke spasticity, a common sequelae of upper motor neuron lesions, results in motor control deficits and pathological hypertonia that not only reduce patients' activities of daily living but may also cause impairment of adaptive neuroplasticity. Repetitive peripheral magnetic stimulation (rPMS), as a novel non-invasive neuromodulation technique, demonstrates unique clinical potential through targeted modulation of electromagnetic coupling effects in the peripheral neuromuscular system. Although current international studies have validated the therapeutic potential of rPMS for spasticity, significant heterogeneity persists in elucidating its mechanisms of action, optimizing parameter protocols, and standardizing outcome assessment systems. This review innovatively synthesized recent randomized controlled trials (RCTs) and mechanistic evidence, systematically summarizing rPMS-mediated multidimensional intervention paradigms for upper- and lower-limb spasticity. It rigorously examined the correlations between stimulation frequency parameters (low-frequency vs. high-frequency), anatomical targeting (nerve trunk vs. motor point), and clinical outcomes including spasticity severity, motor function, and quality of life. Crucially, the analysis reveals that rPMS may ameliorate spasticity after stroke through dual mechanisms involving local neuroelectrophysiological modulation and central sensorimotor network reorganization, thereby providing a theoretical foundation for developing individualized rPMS clinical protocols and establishing precision treatment strategies.
Humans
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Lower Extremity/physiopathology*
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Upper Extremity/physiopathology*
;
Muscle Spasticity/physiopathology*
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Stroke/physiopathology*
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Magnetic Field Therapy/methods*
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Spasm/etiology*
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Stroke Rehabilitation
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Randomized Controlled Trials as Topic
9.Chemical knockdown of Keap1 and homoPROTAC-ing allergic rhinitis.
Jianyu YAN ; Tianyu WANG ; Ruizhi YU ; Lijuan XU ; Hongming SHAO ; Tengfei LI ; Zhe WANG ; Xudong CHA ; Zhenyuan MIAO ; Chengguo XING ; Ke XU ; Huanhai LIU ; Chunlin ZHUANG
Acta Pharmaceutica Sinica B 2025;15(8):4137-4155
Allergic rhinitis (AR), a globally prevalent immune-mediated inflammatory condition, is still an incurable disease. In the present study, we have validated the impact of the Kelch-like ECH associated protein 1 (Keap1)-related oxidative stress and inflammatory response in clinical AR patient peripheral blood and nasal swab samples, emphasizing the biological relevance of Keap1 and AR. Targeting Keap1 -nuclear factor erythroid 2-related factor 2 (Nrf2) related anti-oxidative stress may be effective for AR intervention. Drawing inspiration from the Keap1 homodimerization and the E3 ligase characteristics, we herein present a design of novel bivalent molecules for chemical knockdown of Keap1. For the first time, we characterized ternary complexes of Keap1 dimer and one molecule of bivalent compounds. The best bivalent molecule 8 encompasses robust capacity to degrade Keap1 as a homoPROTACKEAP1. It efficaciously suppresses inflammatory cytokines in extensively different cells, including human nasal epithelial cells. Moreover, in an AR mouse model, we confirmed that the chemical degradation induced by homoPROTACKEAP1 led to therapeutic benefits in managing AR symptoms, oxidative stress and inflammation. In summary, our findings underscore the efficacy of targeting the Keap1 system through the homoPROTAC-ing technology as an innovative and promising treatment strategy for the incurable allergic disorders.
10.Mechanisms of Neiyiting Decoction in Preventing Postoperative Recurrence of Endometriosis by Inhibiting Macrophage M1 Polarization Through the TREM1/TLR4/NF-κB Signaling Pathway
Lijuan LU ; Weiwei ZENG ; Jingwen WANG ; Lurong ZHANG ; Ying LIU ; Xueliang SUN ; Wen XU ; Xiaohui FANG ; Yichao YAN ; Qinhua XIA
Journal of Sichuan University (Medical Sciences) 2025;56(2):371-381
Objective The high post-surgery recurrence rate of endometriosis(EMs)has emerged as a challenge in the long-term manaagement of the condition.This study is aimed at investigating the mechanisms of Neiyiting(NYT)decoction in preventing postoperative recurrence of EMs.Methods An animal model of EMs postoperative recurrence and a model of endometrial stromal cells(hEM15A)cocultured with macrophages(RAW 264.7 cell line)were established for both in vivo and in vitro experiments.An autotransplantation method was used to establish a rat model of EMs.The rats were divided into 4 groups(6 rats per group)and received the corresponding treatments:a Model group receiving distilled water,a Gestrinone group receiving gestrinone at 0.325 mg/kg,a low-dose NYT(NYT-L)group receiving NYT decoction at 5.04 g/(kg-d),and a high-dose NYT(NYT-H)group receiving NYT decoction at 10.08 g/(kg-d).The treatment was administered for 3 weeks via intragastric gavage.In addition,6 SD rats were randomly selected for the control group(Control group),and were given distilled water for 3 weeks via intragastric gavage.The sizes and pathological changes of recurrent lesions in EMs rats were observed.Immunohistochemistry and qRT-PCR were performed to assess the expression of M1 macrophage marker CD86 protein and mRNA in vivo.Additionally,immunohistochemistry and qRT-PCR were used to assess the expression of indicator proteins related to the triggering receptor expressed on myeloid cells 1(TREM1)/Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)signaling pathway and mRNA.The proliferation of hEM15A cells in the coculture experiment was observed.Flow cytometry was performed to determine the polarization of RAW264.7 macrophages,and qRT-PCR was used to determine the expression levels of inducible nitric oxide synthase(iNOS)and interleukin 1β(IL-1β)mRNA.Western blot was performed to determine the expression of signaling pathway-related indicator proteins in vitro.ELISA was performed to determine the levels of inflammatory factors in vitro.Results Compared with the Model group,the volume of recurrent lesions in the NYT-H group was reduced(P<0.01).Findings from the macrophage M1 polarization assessment showed that the expression levels of CD86 protein and mRNA in the recurrent lesions of the Model group were higher than those in the control group(P<0.01).The expression levels of CD86 protein and mRNA in the recurrent lesions of the NYT-H group were lower than those of the Model group(P<0.01).In addition,the RAW 264.7 cell experiment further verified that NYT decoction could reduce the number of CD86-positive macrophages induced by plasmids overexpressing TREM1 and reduce the expression of IL-1β and iNOS mRNA(P<0.01).The results of the hEM15A cell proliferation assay showed that NYT decoction down-regulated KI-67 protein expression in hEM15A cells induced by macrophage M1 polarization(P<0.01).The results of TREM1/TLR4/NF-κB signaling pathway showed that the protein and mRNA expression levels of TREM1,TLR4,and NF-κB in the recurrent lesions of the Model group were higher than those of the control group(P<0.01).Compared with those in the Model group,the protein and mRNA expression levels of TREM1,TLR4,and NF-κB in the recurrent lesions of the NYT-H group were lower(P<0.01).In addition,the coculture experiment of RAW264.7 and hEM15A cells further confirmed that NYT decoction reduced the expression of TREM1,TLR4,and P-P65 proteins(P<0.01).Conclusion NYT decoction can inhibit macrophage M1 polarization through the TREM1/TLR4/NF-κB signaling pathway,improve the inflammation level,and inhibit the formation of ectopic endometrial lesions,thereby preventing postoperative recurrence of EMs.


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