1.Endothelial progenitor cell and mesenchymal stem cell therapy for vascular stent-associated diseases
Qingyin LI ; Linhua LI ; Chunle ZHANG ; Ping FU
Chinese Journal of Tissue Engineering Research 2025;29(19):4091-4101
BACKGROUND:With advancements in stem cell research,the therapeutic efficacy of adult stem cells such as endothelial progenitor cells and mesenchymal stem cells in atherosclerosis and complications arising from atherosclerosis and vascular stent implantation is gradually being recognized.Due to the limitations of intravenous infusion of adult stem cells,including poor targeting and low treatment efficiency,recent research has focused on surface modification of vascular stents to achieve localized aggregation and functional modulation of endothelial progenitor cells or mesenchymal stem cells. OBJECTIVE:To discuss the therapeutic progress of endothelial progenitor cells and mesenchymal stem cells in vascular stent-related diseases,summarize the research status of the design of vascular stents based on endothelial progenitor cells and mesenchymal stem cells. METHODS:Relevant literature was retrieved on CNKI,WanFang,PubMed,and Web of Science databases since their inception.The Chinese search terms were"endothelial injury,stenting,thrombosis,intimal hyperplasia,atherosclerosis,endothelial repair,endothelial progenitor cell,mesenchymal stem cell,vascular stent."English search terms were"endothelial injury,stenting,thrombosis,intimal hyperplasia,atherosclerosis,endothelial repair,endothelial regeneration,endothelial progenitor cell,mesenchymal stem cell,vascular stent,vascular scaffold."According to inclusion and exclusion criteria,127 articles were finally reviewed. RESULTS AND CONCLUSION:Endothelial progenitor cells and mesenchymal stem cells can treat atherosclerosis and complications of stent implantation through differentiation and paracrine effects,mainly by protecting endothelial cells,regulating the expression of inflammatory cells and cytokines,and modulating smooth muscle cell proliferation and phenotype.Mesenchymal stem cells may have adverse reactions such as thrombosis and vascular calcification in therapeutic applications,and using extracellular vesicles and co-administration with heparin for surface modification is a feasible solution.Currently,there is more research on stents based on endothelial progenitor cells,mainly focusing on recruitment,capture,proliferation,differentiation,and activity.Research on stents based on mesenchymal stem cell capture in the vascular field is relatively scarce,but exosome-loaded stents derived from mesenchymal stem cells have been found to have highly effective therapeutic efficacy.Additionally,some underlying diseases such as diabetes may affect the activity of adult stem cells,leading to the loss of effectiveness in stem cell-based stent designs.Therefore,in future stent designs,consideration should be given to the background diseases.
2.Effects of Different Modes in Hypoxic Training on Metabolic Improvements in Obese Individuals: a Systematic Review With Meta-analysis on Randomized Controlled Trail
Jie-Ping WANG ; Xiao-Shi LI ; Ru-Wen WANG ; Yi-Yin ZHANG ; Feng-Zhi YU ; Ru WANG
Progress in Biochemistry and Biophysics 2025;52(6):1587-1604
This paper aimed to systematically evaluate the effects of hypoxic training at different fraction of inspired oxygen (FiO2) on body composition, glucose metabolism, and lipid metabolism in obese individuals, and to determine the optimal oxygen concentration range to provide scientific evidence for personalized and precise hypoxic exercise prescriptions. A systematic search was conducted in the Cochrane Library, PubMed, Web of Science, Embase, and CNKI databases for randomized controlled trials and pre-post intervention studies published up to March 31, 2025, involving hypoxic training interventions in obese populations. Meta-analysis was performed using RevMan 5.4 software to assess the effects of different fraction of inspired oxygen (FiO2≤14% vs. FiO2>14%) on BMI, body fat percentage, waist circumference, fasting blood glucose, insulin, HOMA-IR, triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C), with subgroup analyses based on oxygen concentration. A total of 22 studies involving 292 participants were included. Meta-analysis showed that hypoxic training significantly reduced BMI (mean difference (MD)=-2.29,95%CI: -3.42 to -1.17, P<0.000 1), body fat percentage (MD=-2.32, 95%CI: -3.16 to -1.47, P<0.001), waist circumference (MD=-3.79, 95%CI: -6.73 to -0.85, P=0.01), fasting blood glucose (MD=-3.58, 95%CI: -6.23 to -0.93, P=0.008), insulin (MD=-1.60, 95%CI: -2.98 to -0.22, P=0.02), TG (MD=-0.18, 95%CI: -0.25 to -0.12, P<0.001), and LDL-C (MD=-0.25, 95%CI: -0.39 to -0.11, P=0.000 3). Greater improvements were observed under moderate hypoxic conditions with FiO2>14%. Changes in HOMA-IR (MD=-0.74, 95%CI: -1.52 to 0.04,P=0.06) and HDL-C (MD=-0.09, 95%CI: -0.21 to 0.02, P=0.11) were not statistically significant. Hypoxic training can significantly improve body composition, glucose metabolism, and lipid metabolism indicators in obese individuals, with greater benefits observed under moderate hypoxia (FiO>14%). As a key parameter in hypoxic exercise interventions, the precise setting of oxygen concentration is crucial for optimizing intervention outcomes.
3.Mechanism of Huangqi Guizhi Wuwutang in Treatment of Sarcopenia Associated with Rheumatoid Arthritis by Improving Skeletal Muscle Homeostasis Through Regulation of Autophagy
Yakun WAN ; Yuan LIU ; Yuan QU ; Jingyu GUO ; Ting LIU ; Zhihui BAI ; Di ZHANG ; Ping JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):12-23
ObjectiveThis study aims to explore the mechanism of action of Huangqi Guizhi Wuwutang in treating rheumatoid arthritis (RA)-associated sarcopenia by regulating autophagy and improving skeletal muscle homeostasis based on network pharmacology,bioinformatics,machine learning,and animal experiments. MethodsActive ingredients and targets of Huangqi Guizhi Wuwutang were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP),PubChem,and SwissTargetPrediction databases. RA-related datasets were retrieved from the GEO database,and differential genes were screened. Sarcopenia-related targets were searched through GeneCards and the Comparative Toxicology Database (CTD),and autophagy-related gene sets were downloaded from the Human Autophagy Database (HADb). Their intersection was analyzed to identify autophagy-related therapeutic targets,followed by enrichment analysis. A protein-protein interaction (PPI) network was constructed using the STRING database,and key targets were selected using multiple methods. Machine learning was applied to predict models based on the expression profiles of intersecting targets,and nomogram models were constructed based on key targets. Molecular docking of the top four active ingredients with key targets was performed using AutoDockVina. A collagen-induced arthritis (CIA) rat model was established using bovine type Ⅱ collagen,with SD rats divided into groups including a blank group,a model group,and low-,medium-,and high-dose groups of Huangqi Guizhi Wuwutang (2.44,4.88,and 9.76 g·kg-1) and administered for five consecutive weeks. Joint scores and gastrocnemius muscle mass were recorded and analyzed after modeling. Hematoxylin and eosin (HE) staining and Masson's staining were used to observe pathological changes in muscle tissue. Immunofluorescence staining was applied to observe the protein expression levels of myosin heavy chain (MYHC) and insulin-like growth factor-1 (IGF-1) in skeletal muscle. Western blot was used to detect the protein expression levels of autophagy-related proteins ATG5,Beclin1,LC3B,muscle-specific proteins (MuRF1),MaFbx,and MYHC. Real-time quantitative reverse transcription PCR (Real-time PCR) was performed to measure the mRNA expression levels of ATG5,Beclin1,LC3B,MuRF1,MaFbx,and MYHC in muscle tissue. ResultsNetwork pharmacology revealed that Huangqi Guizhi Wuwutang shared 25 common targets with autophagy genes related to RA-associated sarcopenia. The PPI network and machine learning identified six key targets,which were primarily involved in autophagy and inflammatory pathways. Animal experiments showed that compared to the blank group,the model group had significantly higher joint scores (P<0.01) and lower gastrocnemius muscle index (P<0.01). HE staining indicated a significant reduction in the cross-sectional area of gastrocnemius muscle fibers,with notable inflammatory cell infiltration and muscle atrophy in the model group. Masson's staining revealed obvious collagen fiber proliferation and deposition,with significant muscle fibrosis in the model group. The protein and mRNA expression levels of ATG5,Beclin1,LC3B,MuRF1,and MaFbx were significantly increased (P<0.01),while the protein expression of MYHC and IGF1 was significantly downregulated (P<0.01). Compared with the model group,the high-dose group of Huangqi Guizhi Wuwutang showed significantly reduced protein and mRNA expression levels of ATG5,Beclin1,LC3B,MuRF1,and MaFbx (P<0.01) and increased protein expression levels of MYHC and IGF1 (P<0.01). The cross-sectional area of muscle fibers increased,and the muscle cell morphology approached normal. Moreover,pathological abnormalities in the gastrocnemius muscle were significantly improved,with reduced collagen fiber proliferation (P<0.01). ConclusionHuangqi Guizhi Wuwutang can mediate autophagy by regulating the expression of ATG5,Beclin1,LC3B,and IGF1,thereby reducing skeletal muscle catabolism and improving skeletal muscle homeostasis,which contributes to the treatment of RA-associated sarcopenia. The findings provide insight into the mechanisms underlying the effects of Huangqi Guizhi Wuwutang in the treatment of RA-related sarcopenia and offer a reference for its enhanced clinical application.
4.Association of sleep quality with overweight and obesity among middle school students
Chinese Journal of School Health 2025;46(6):779-782
Objective:
To explore relationship of sleep quality with overweight and obesity among middle school students, so as to provide a reference basis for improving adolescent sleep health.
Methods:
From September to December 2023, 5 713 middle school students aged 13 to 18 were selected by stratified cluster random sampling method in six regions, including Shanghai, Suzhou, Taiyuan, Wuyuan, Xingyi and Urumqi. Sleep quality survey was conducted on middle school students by Pittsburgh Sleep Quality Index. Height and weight were measured, and World Health Organization s standards for growth and development of children and adolescents was used to evaluate their nutritional status. Both χ 2 test and Logistic regression analysis were used to analyze the association between sleep quality and nutritional status of middle school students.
Results:
The non compliance detection rate of sleep quality was 38.4% among girls, but 29.2% among boys, and the difference was of statistical significance( χ 2=54.08, P < 0.01 ). The detection rate of substandard sleep quality was 34.2% in the group with normal nutritional status, 38.3% in the group with overweight, 43.7% in the group with obesity and 26.0% in the group with emaciation, and the difference in the rates of substandard sleep quality among middle school students of different nutritional status was statistically significant ( χ 2=68.15, P <0.01). Logistic regression analysis showed that, after controlling for mental health and physical activity, the detection rate of substandard sleep quality in the obese groups was 1.30 times higher than that in the normal group, respectively( OR =1.30, 95% CI =1.06- 1.59 , P <0.01).
Conclusions
Sleep quality is correlated with overweight and obesity among middle school students, and there are gender differences. Intervention policies should be formulated according to the characteristics of different genders.
5.Analysis of pollution of PM 2.5 in children s bedrooms caused by using solid fuels and the influencing factors
ZHENG Ping, SHI Chunli, XIN Shuzhi, CHEN Shunqiang, SHEN Yue, ZHANG Bei, XU Ning, WANG Qiang
Chinese Journal of School Health 2025;46(7):932-936
Objective:
To investigate the indoor fine particulate matter (PM 2.5 ) pollution and its influencing factors in children s bedrooms using solid fuel, so as to provide evidence for effective strategy to reduce PM 2.5 pollution.
Methods:
From December 2019 to November 2020, 198 households (108 in the north, 90 in the south) from two pilots in the north(Jiamusi in Heilongjiang Province) and south of China (Mianyang in Sichuan Province) were selected, and status of solid fuels using were obtained through home visits, dynamic changes in PM 2.5 concentrations in children s bedrooms were monitored by using real time online instruments, and the influencing factors of PM 2.5 pollution were analyzed by using a mixed effects model.
Results:
During the monitoring period, the daily PM 2.5 concentrations in the northern and southern pilot were 78.33 (40.50, 154.80) and 38.54(26.20, 58.46) μg/m 3, respectively, exceeding standard rates of 44.57% and 33.22%. During the heating period, the daily PM 2.5 concentrations in the northern and southern pilot were 212.50(133.60,244.10) and 104.42(73.97, 134.90) μg/m 3, respectively, with over standard rates of 96.75% and 86.96%. The mixed effects model analysis results showed that children s bedroom PM 2.5 concentrations were associated with solid fuel usage duration, window opening time, room layout (shared entrance door between kitchen and bedroom), indoor smoking, indoor humidity, and solid fuel use in the bedroom ( β =0.19, -0.05, 1.20, 0.43, 0.02, 0.35, all P <0.05).
Conclusion
Solid fuel combustion significantly comtributes to PM 2.5 pollution in children s bedrooms, with more pronounced impacts observed in northern China compared to southern regions.
6.Correlation between heart rate variability and extracurricular physical exercise of primary and middle school students in Chengdu City
PAN Zhongjin, ZHANG Yihong, HE Zhongtao, LIU Jianyu, ZHENG Xiao, SHAO Ping
Chinese Journal of School Health 2025;46(7):961-964
Objective:
To investigate the impact of extracurricular physical activity on heart rate variability (HRV) among primary and secondary school students in Chengdu City, so ao to provide references for scientific exercise prescription.
Methods:
Using a convenient sampling method, 1 323 primary and secondary students were enrolled from central Chengdu who underwent physical fitness assessments at Sichuan Provincial Institute of Sports Science from September 2020 to January 2022. According to the standards of the National Physical Fitness Monitoring Center, boys and girls were divided into groups with and without extracurricular physical exercise habits. HRV was monitored using the SA-3000P device. Key HRV parameters were evaluated separately by gender, including standard deviation of normal to normal intervals (SDNN), root mean square of successive differences (rMSSD), total power (TP), low frequency power (LF) and high frequency power (HF). Statistical analyses were employed by t-test, Mann-Whitney U-test and Chi square test.
Results:
lgSDNN, lgrMSSD, TP, LF and HF in the group without extracurricular physical exercise habit [boys:(1.67±0.13)ms, (1.59±0.20)ms, (7.34±0.73)ms 2, (6.11±0.74)ms 2, (6.05±0.87)ms 2; girls:(1.67± 0.13)ms , (1.59±0.19)ms, (7.35±0.60)ms 2, (6.06±0.69)ms 2, (6.12±0.87)ms 2] were lower than those in the group with extracurricular physical exercise habit [boys:(1.75±0.13)ms, (1.72±0.18)ms, (7.69±0.62)ms 2, (6.41±0.76)ms 2, (6.44±0.79)ms 2;girls:(1.73±0.13)ms, (1.68±0.20)ms, (7.60±0.65)ms 2, (6.26±0.86)ms 2, (6.36±0.90)ms 2] ( t =-8.24, -8.75, -6.54, -5.35 , -6.33;-5.10,-4.90,-4.47,-2.71,-2.93, all P <0.01). Only the group of boys without extracurricular physical exercise habits showed a decrease in lgLF/HF [0.04(-0.19,0.27)] compared to the group with extracurricular physical exercise habits [ -0.03 (-0.25,0.20)] ( Z=-2.01, P <0.05). When the score classes of autonomic nerve activity, stress index and fatigue index were compared between boys and girls groups without and with extracurricular physical exercise habits, the proportion of boys normal and above scores increased from 79.3%, 84.1%, 71.8% to 91.4%, 95.7%, 87.3%, respectively, and the differences were statistically significant ( χ 2=47.42, 63.66, 38.28); the proportion of girls normal score and above increased from 79.8%, 85.7%, 75.0% to 85.4%, 92.1%, 79.4%, and the differences were statistically significant ( χ 2=48.31, 22.18, 22.22) (all P <0.01).
Conclusion
The primary and secondary school students who have the habit of extracurricular physical exercise have enhanced compliance in indicators related to HRV, showing more complex heart rate variability.
7.Treatment of Psoriasis from ''Fuyu'' Theory
Jiaqi LI ; Bobiao NING ; Ningxin ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):39-48
Psoriasis is a chronic inflammatory skin disease with a polygenic genetic background. Its etiology remains unclear, and its pathogenesis is complex and refractory, collectively posing significant challenges in its treatment and greatly affecting the physical and mental health of patients. With the advantages of multi-target and multi-pathway treatment, traditional Chinese medicine has shown unique efficacy and value in the diagnosis and treatment of psoriasis. Modern doctors have a lot of discussion on psoriasis, and most of them tend to treat the disease by solving disorders of the blood system. They think that the disease is closely related to the "heat in the blood". Combining the clinical characteristics and accompanying symptoms of psoriasis, this article traced the causes of "heat in the blood" in psoriasis and believed that multiple internal and external factors have prevented the smooth circulation of Qi. Yang Qi Fuyu (stagnation) and transformation into heat and toxicity is the source of "heat in the blood" in psoriasis. Furthermore, it was proposed that "Fuyu" is the core pathogenesis of psoriasis. The etiology of "Fuyu" is complex, such as external wind and cold pathogens, emotional injuries, internal accumulation of dampness, and deficiency of healthy Qi, all of which can disrupt the ascending and descending movement of Qi, impede the circulation of Qi and fluids, close the pores and skin texture, and subsequently lead to stagnation. Based on the above understanding, "resolving stagnation" is crucial for treating the disease. Many doctors have explored the treatment ideas of psoriasis from the perspectives of dispelling wind, warming cold, regulating Qi, eliminating dampness, tonifying deficiency, and external treatment, aiming to remove the causes, promote the circulation of Qi and fluids, and resolve stagnation and heat. Clinical studies have shown that the therapies can relieve clinical symptoms, reduce recurrence rate, and improve quality of life, which also have good safety in the treatment of psoriasis. This article discussed the treatment of psoriasis from the perspective of "Fuyu", enriching the understanding of TCM regarding the etiology and pathogenesis of psoriasis. It is aiming to serve as an effective supplement to the "treating by solving disorders of the blood system" approach and provide a reference for clinical diagnosis and treatment of psoriasis.
8.Exploration of Kaixuan Jiedu Core Prescription's Efficacy in Alleviating Psoriasis Through Modulation of Ferroptosis Pathways: An Integrative Approach Involving Bioinformatics and Experimental Validation
Haoruo YANG ; Xue XIAO ; Jiaqi LI ; Ningxin ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):69-78
ObjectiveTo use bioinformatics technology to screen the molecular patterns and diagnostic biomarkers of ferroptosis closely related to psoriasis, observe the therapeutic effect of Kaixuan Jiedu core prescription on psoriasis and explore its potential mechanism through animal experiments. MethodsPsoriasis microarray data from GEO were analyzed to identify differentially expressed genes (DEGs). Intersection with a ferroptosis gene set yielded psoriasis ferroptosis-related genes (FRGs), which underwent correlation, consensus clustering, enrichment, and immune infiltration analyses. Core diagnostic FRGs (Hub-FRGs) were identified using random forest (RF), support vector machine (SVM), LASSO regression, Nomogram, and ROC analyses. In vivo, imiquimod (5% cream) induced psoriasis in mice (except controls). Drug treatment groups received respective doses, while control and model groups received saline via daily gavage for 7 days. Back skin changes were recorded and PASI scored. Hematoxylin-eosin (HE) staining assessed histopathology. The levels of ferrous ion (Fe2+), malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) and free fatty acid (FFA) in skin tissue were detected. The level of reactive oxygen species (ROS) in skin tissue was detected by immunofluorescence. Immunohistochemistry was used to detect the expression of ChaC glutathione-specific γ-glutamyl transferase 1 (CHAC1), arachidonic acid 12-lipoxygenase β (ALOX12B), trimotif protein 21 (TRIM21), proliferation marker (Ki67) and nuclear transcription factor-κB (NF-κB) protein. ResultsAnalysis of GSE30999 identified 2 100 DEGs and 24 FRGs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment revealed 1 000 biological functions and 75 pathways. After cluster analysis, combined with three machine learning algorithms, Nomogram and ROC curve analysis, the core Hub-FRGs (CHAC1, ALOX12 B, TRIM21) were obtained. Immunoinfiltration showed inactive memory CD4+T cells and activated dendritic cells abundance significantly correlated with Hub-FRGs. In vivo, model group vs. control showed significantly increased PASI/Baker scores (P<0.05), epidermal hyperkeratosis, inflammatory infiltration, and elevated levels of Fe2+, MDA, 4-HNE, FFA, ROS, CHAC1, ALOX12B, TRIM21, Ki67, and NF-κB (P<0.05). Drug groups vs. model group exhibited significantly reduced scores (P<0.05), alleviated skin lesions, and decreased levels of Fe2+, MDA, 4-HNE, FFA, ROS, Hub-FRGs, Ki67, and NF-κB (P<0.05). ConclusionKaixuan Jiedu core prescription can significantly improve the skin pathological injury of psoriasis mice, showing good therapeutic and repair effects, and its mechanism may be related to regulating the expression of ferroptosis genes CHAC1, ALOX12B and TRIM21, which are closely related to the pathogenesis of psoriasis.
9.Kaixuan Jiedu Compatibility Alleviates Skin Lesions and Inflammatory Reactions in Psoriasis-like Mice
Ningxin ZHANG ; Jiaqi LI ; Xinqian LIU ; Tianbo ZHANG ; Meiqi SUN ; Mingjing LI ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):79-88
ObjectiveTo investigate the efficacy and safety of Kaixuan Jiedu compatibility and the decomposed prescriptions in the treatment of psoriasis. MethodsThirty Balb/c mice were randomly grouped as follows (n=6): normal, model, Kaixuan Jiedu (KXJD, 15.21 g·kg-1), Kaixuan (KX, 3.08 g·kg-1), and Jiedu (JD, 12.13 g·kg-1). Except the normal group, the rest groups were modeled for psoriasis-like skin lesions by topical application of imiquimod, and samples were collected after 7 days of continuous intervention. Mice were photographed at the lesion site during modeling and before sampling and the psoriasis area and severity index (PASI) was calculated. Hematoxylin-eosin (HE) staining was used to observe pathological changes in the lesions and measure the epidermal thickness. Mice were photographed and observed for the tortuous dilation of dermal capillaries. The expression of vascular endothelial growth factor (VEGF), platelet-endothelial cell adhesion molecule (CD31), proliferating cell nuclear antigen (Ki67), and cytokeratin 10 (CK10) in the epidermal tissue was detected by immunohistochemistry. Immunofluorescence assay was employed to determine the expression of Claudin-1 and Occludin. Real-time PCR was employed to determine the mRNA levels of interleukin-17A (IL-17A) and interleukin-23 (IL-23). The spleen and thymus were photographed and weighed, and the spleen and thymus indices were calculated. The safety of the treatment was assessed by automatic biochemistry testing of the serum, liver, and kidney functions and by HE staining of the liver, kidney and spleen. ResultsCompared with that of the normal group, the skin of the model group showed erythema, infiltration, and typical psoriasis-like changes, tortuous dilation of dermal capillaries, hyperkeratosis in epidermal cells, acanthosis, massive lymphocytic infiltration in the dermis, impaired barrier function, increased expression of VEGF, CD31, Ki67, and CK10 (P<0.01), reduced expression of Claudin-1 and Occludin (P<0.01) in the epidermis, and up-regulated mRNA levels of IL-17A and IL-23 (P<0.01). In addition, the mice in the model group showed spleen enlargement, thymus atrophy, increased spleen index, and decreased thymus index (P<0.01). Compared with the model group, KXJD and JD reduced psoriasis-like skin lesions, inhibited the tortuous dilation of dermal capillaries, reduced the expression of VEGF, CD31, Ki67, and CK10 (P<0.01), increased the expression of claudin-1 (P<0.01), and down-regulated the mRNA levels of inflammatory factors (P<0.01). Moreover, the KXJD group outperformed the JD group. The JD group showed no significant difference from the model group regarding the spleen index, thymus index, and Occludin expression. The psoriasis indicators in the KX group were not significantly different from those in the model group. ConclusionKXJD and JD can reduce the symptoms of local skin lesions of psoriasis, which is manifested as different inhibition degrees of the proliferation and differentiation of keratin-forming cells, tortuous dilation of dermal capillaries, and inflammatory reactions, as well as the protection of the skin barrier. Moreover, KXJD outperformed JD. KX alone did not significantly reduce psoriasis lesions in mice. KXJD and the decomposed prescriptions are safe and effective, causing no obvious liver and kidney injuries.
10.Mechanism of Neochlorogenic Acid in Ameliorating Psoriatic Keratinocyte Proliferation and Inflammation by Targeting HSP90 to Modulate NF-κB/NLRP3 Signaling Pathway
Mengyao JIANG ; Xinwei ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):89-98
ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid (NA) and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe (NA-P) and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8(CCK-8). The potency of NA and NA-P was evaluated in the safe concentration range, and the effects of 0-100 μmol·L-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-23 (IL-23), and interleukin-17A (IL-17A) inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA), and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to measure the effects of NA on the mRNA expression of keratin 16 (K16) in HaCaT cells, S100 calcium-binding protein A9 (S100A9), S100 calcium-binding protein A7 (S100A7), IL-6, IL-17A, and chemokine 1 (CXCL1). In vitro fluorescence labeling and competition experiments using NA-P were performed, and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry (Pull-down/LC-MS/MS) were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting (Pull down-WB), cellular heat transfer analysis combined with protein immunoblot (CETSA-WB) experiments, and molecular docking. Finally, Real-time PCR was utilized to detect the effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the mRNA expression of IL-1β, nucleotide-binding oligomeric structural domain-like receptor protein 3 (NLRP3), apoptosis-associated speckled-like protein (ASC), and cysteine protease-1 (Caspase-1) in HaCaT cells. Protein immunoblot (Western blot) was used to detect the effects of phosphorylated p65 (p-p65), p65, phosphorylated human nuclear factor-κB inhibitory protein α (p-IκBα), human nuclear factor κB inhibitory protein α (IκBα), and heat shock protein 90 (HSP90) expression. ResultsIn the 200 μmol·L-1 safe concentration range, HaCaT cell proliferation, increased expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors, and increased mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L-1 NA and NA-P groups was inhibited, and the expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors was decreased in the NA-L, NA-M, NA-H, and NA-P-H groups. The mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 was decreased (P<0.05). High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L-1 NA competing with 100 μmol·L-1 NA-P. Compared with that in the blank group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1, as well as the expression of p-p65/p65, p-IκBα/IκBα, and HSP90 protein, were increased in HaCaT cells in the M5 group (P<0.05). Compared with that in the M5 group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1 of cells in the NA-L group, the NA-M group, the NA-H group, and the NA-P-H group was decreased (P<0.05). p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups (P<0.05), and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein, and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1, HSP90B1, and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.


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