1.Construction and in vitro osteogenic activity study of magnesium-strontium co-doped hydroxyapatite mineralized collagen
WANG Meng ; SUN Yifei ; CAO Xiaoqing ; WEI Yiyuan ; CHEN Lei ; ZHANG Zhenglong ; MU Zhao ; ZHU Juanfang ; NIU Lina
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(1):15-28
Objective:
To investigate the efficacy of magnesium-strontium co-doped hydroxyapatite mineralized collagen (MSHA/Col) in improving the bone repair microenvironment and enhancing bone regeneration capacity, providing a strategy to address the insufficient biomimetic composition and limited bioactivity of traditional hydroxyapatite mineralized collagen (HA/Col) scaffolds.
Methods:
A high-molecular-weight polyacrylic acid-stabilized amorphous calcium magnesium strontium phosphate precursor (HPAA/ACMSP) was prepared. Its morphology and elemental distribution were characterized by high-resolution transmission electron microscopy (TEM) and energy-dispersive spectroscopy. Recombinant collagen sponge blocks were immersed in the HPAA/ACMSP mineralization solution. Magnesium-strontium co-doped hydroxyapatite was induced to deposit within collagen fibers (experimental group: MSHA/Col; control group: HA/Col). The morphological characteristics of MSHA/Col were observed using scanning electron microscopy (SEM). Its crystal structure and chemical composition were analyzed by X-ray diffraction and Fourier transform infrared spectroscopy, respectively. The mineral phase content was evaluated by thermogravimetric analysis. The scaffold's porosity, ion release, and in vitro degradation performance were also determined. For cytological experiments, CCK-8 assay, live/dead cell staining, alkaline phosphatase staining, alizarin red S staining, RT-qPCR, and western blotting were used to evaluate the effects of the MSHA/Col scaffold on the proliferation, viability, early osteogenic differentiation activity, late mineralization capacity, and gene and protein expression levels of key osteogenic markers [runt-related transcription factor 2 (Runx2), collagen type Ⅰ (Col-Ⅰ), osteopontin (Opn), and osteocalcin (Ocn)] in mouse embryonic osteoblast precursor cells (MC3T3-E1).
Results:
HPAA/ACMSP appeared as amorphous spherical nanoparticles under TEM, with energy spectrum analysis showing uniform distribution of carbon, oxygen, calcium, phosphorus, magnesium, and strontium elements. SEM results of MSHA/Col indicated successful complete intrafibrillar mineralization. Elemental analysis showed the mass fractions of magnesium and strontium were 0.72% (matching the magnesium content in natural bone) and 2.89%, respectively. X-ray diffraction revealed characteristic peaks of hydroxyapatite crystals (25.86°, 31°-34°). Infrared spectroscopy results showed characteristic absorption peaks for both collagen and hydroxyapatite. Thermogravimetric analysis indicated a mineral phase content of 78.29% in the material. The scaffold porosity was 91.6% ± 1.1%, close to the level of natural bone tissue. Ion release curves demonstrated sustained release behavior for both magnesium and strontium ions. The in vitro degradation rate matched the ingrowth rate of new bone tissue. Cytological experiments showed that MSHA/Col significantly promoted MC3T3-E1 cell proliferation (130% increase in activity at 72 h, P < 0.001). MSHA/Col exhibited excellent efficacy in promoting osteogenic differentiation, significantly upregulating the expression of osteogenesis-related genes and proteins (Runx2, Col-Ⅰ, Opn, Ocn) (P < 0.01).
Conclusion
The MSHA/Col scaffold achieves dual biomimicry of natural bone in both composition and structure, and effectively promotes osteogenic differentiation at the genetic and protein levels, breaking through the functional limitations of pure hydroxyapatite mineralized collagen. This provides a new strategy for the development of functional bone repair materials
2.Arginine Metabolic Disorder in Heart Failure Rats: Analysis Based on Targeted Metabolomics and Bioinformatics
Zeyu LI ; Xiaoqing WANG ; Zhengyu FANG ; Yurou ZHAO ; He XIAO ; Penghaobang LIU ; Haiming ZHANG ; Chunyan LIU ; Yanhong HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):229-237
ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure (HF), providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine (TCM). MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats, which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group, with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-S), and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging (AFADESI-MSI). Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus (GEO), a protein-protein interaction (PPI) network was constructed, and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) was performed. Finally, molecular docking was conducted to verify the binding between core metabolic components and key targets, and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group, the levels of arginine and citrulline in the serum of model rats were significantly decreased (P<0.01), while those of proline, ornithine, creatine, creatinine and glutamate were significantly increased (P<0.05, P<0.01). Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets, such as the angiotensin-converting enzyme (ACE), neuronal nitric oxide synthase (NOS1), 5-hydroxytryptamine receptor 2A (HTR2A), and epidermal growth factor receptor (EGFR), and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway, neuroactive ligand-receptor interactions, and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine, citrulline and HTR2A, as well as between creatine, creatinine and EGFR. Based on pathway-target prediction, potential TCM interventions, such as ginseng and magnolia, were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF, and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways, and offers valuable insights for targeted therapy of HF and the development of TCM.
3.Arginine Metabolic Disorder in Heart Failure Rats: Analysis Based on Targeted Metabolomics and Bioinformatics
Zeyu LI ; Xiaoqing WANG ; Zhengyu FANG ; Yurou ZHAO ; He XIAO ; Penghaobang LIU ; Haiming ZHANG ; Chunyan LIU ; Yanhong HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):229-237
ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure (HF), providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine (TCM). MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats, which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group, with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-S), and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging (AFADESI-MSI). Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus (GEO), a protein-protein interaction (PPI) network was constructed, and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) was performed. Finally, molecular docking was conducted to verify the binding between core metabolic components and key targets, and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group, the levels of arginine and citrulline in the serum of model rats were significantly decreased (P<0.01), while those of proline, ornithine, creatine, creatinine and glutamate were significantly increased (P<0.05, P<0.01). Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets, such as the angiotensin-converting enzyme (ACE), neuronal nitric oxide synthase (NOS1), 5-hydroxytryptamine receptor 2A (HTR2A), and epidermal growth factor receptor (EGFR), and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway, neuroactive ligand-receptor interactions, and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine, citrulline and HTR2A, as well as between creatine, creatinine and EGFR. Based on pathway-target prediction, potential TCM interventions, such as ginseng and magnolia, were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF, and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways, and offers valuable insights for targeted therapy of HF and the development of TCM.
4.Research progress on strategies for toxicity reduction and efficacy enhancement of triptolide
Xiaoqing ZHENG ; Ying DING ; Shanshan XU ; Long WANG ; Shanshan HAN ; Yaping XING ; Meng ZHANG ; Wenhao LI
China Pharmacy 2026;37(11):1496-1501
Triptolide (TP), the core active component of the traditional Chinese medicine Tripterygium wilfordii , exhibits remarkable pharmacological activities including anti-inflammatory, immunosuppressive and anti-tumor effects, and holds broad application prospects in the treatment of major diseases such as autoimmune diseases and malignant tumors. However, TP has a narrow therapeutic window and causes multi-organ toxicities including liver, kidney and reproductive toxicities, which severely restrict its safe clinical application and new drug development. Therefore, toxicity reduction and efficacy enhancement has become a core scientific problem urgently to be solved in this field. This paper systematically reviews the four core strategies for TP toxicity reduction and efficacy enhancement, including structural modification, dosage form improvement, herbal compatibility, and external therapies of traditional Chinese medicine. Among them, structural modification optimizes the toxic and efficacy characteristics of TP from the molecular structure level, with typica l derivatives including (5 R )-5-hydroxy triptolide, ZT01, PG490-88, etc. Dosage form modification achieves toxicity reduction and efficacy enhancement via targeted and sustained-controlled drug release of diverse delivery systems. It includes triptolide preparations such as nanoparticles, liposomes, microemulsion gels and liquid crystals, possessing favorable clinical transformation potential. The herbal compatibility and external therapies of traditional Chinese medicine conform to the holistic view of traditional Chinese medicine and have a profound clinical application foundation, but their mechanisms of action are insufficiently elucidated, and they lack unified standardized specifications and high-quality evidence-based proof. In the future, we should rely on multi-omics technology to elucidate the toxic and efficacy mechanisms, integrate technologies to optimize preparations, improve the evaluation system and promote clinical transformation.
5.Diagnostic value of MS score in macrophage activation syndrome associated with systemic juvenile idiopathic arthritis
Lingling GENG ; Yue PENG ; Duomei SHI ; Li WANG ; Xianyan TANG ; Xinran WEN ; Wenhua ZHANG ; Xiaoqing LI
International Journal of Pediatrics 2025;52(7):476-480
Objective:To explore the diagnostic value of the macrophage activation syndrome/systemic juvenile idiopathic arthritis(MS)score in macrophage activation syndrome(MAS)associated with systemic juvenile idiopathic arthritis(sJIA),and to provide a reference for clinical work.Methods:This study was a retrospective case-control analysis,conducted on the patients initially diagnosed as sJIA-associated with MAS and admitted into the Department of Rheumatology and Immunology of Children's Hospital Affiliated to Xi 'an Jiaotong University from July 1st,2016 to June 30th,2023. All of the patients met the diagnostic criteria for patients with MAS associated with sJIA according to the 2016 European Alliance of Associations for Rheumatology(EULAR)/American College of Rheumatology(ACR)/Pediatric Rheumatology International Trials Organization(PRINTO)standards. The basic information at baseline,clinical manifestations,and auxiliary examination results were collected. The MS score was applied to re-evaluate the children diagnosed as sJIA-associated with MAS. When the MS score ≥-2.1,the possibility of sJIA with MAS was high. Thirty cases of sJIA without MAS were randomly selected as the control group.Results:There were 28 cases in the MAS group,including 13 males(46.43%)and 15 females(53.57%),with an average age of(7.51±4.01)years. Compared with the control group,the MAS group were significantly more likely to have high fever( χ2=8.539, P=0.003),hepatomegaly( χ2=11.621, P<0.001),splenomegaly( χ2=11.710, P<0.001)and neurological involvement( χ2=27.619, P<0.001),with the differences being statistically significant. Meanwhile,there were statistically significant differences between the two groups in terms of white blood cell count( Z=-4.001, P<0.001),neutrophil count( Z=-3.659, P<0.001),platelet count( Z=-4.687, P<0.001),albumin level( Z=-4.018, P<0.001),alanine aminotransferase( Z=-3.846, P<0.001),aspartate aminotransferase( Z=-5.932, P<0.001),lactate dehydrogenase( Z=-6.150, P<0.001),triglycerides( Z=-5.874, P<0.001),fibrinogen( Z=-5.808, P<0.001),ferritin( Z=-5.280, P<0.001),erythrocyte sedimentation rate( Z=-3.971, P<0.001),ferritin/erythrocyte sedimentation rate( Z=-5.433, P<0.001),reduction of two-line cells in blood( χ2=11.408, P<0.001)and the presence of hemophagocytosis in bone marrow smears( χ2=28.260, P<0.001). Moreover,there was a statistically significant difference in MS scores between the two groups( Z=-6.148, P<0.001),with higher MS scores in the MAS group. Nevertheless,this study showed the median MS scores of both groups ≥-2.1. Conclusion:The MS score was significant to a certain degree as reference for the diagnosis of MAS,and this study showed that the MS score in the MAS group was significantly higher than the control group. However,the median MS scores in both groups were no less than -2.1. This might be related to the influence of factors during the assessment,which made it necessary to optimize the cutoff values of the MS score. Therefore,prospective studies should be carried out on the role of MS score in early identification of MAS.
6.Potential mechanisms of action of Jiao'ai decoction in the treatment of premature ovarian failure: Integrated molecular pathway, gut flora, and untargeted metabolism
Da Zhang ; Xiaoqing Zhang ; Dongsheng Wei ; Ning Wang ; Piwen Zhao
Journal of Traditional Chinese Medical Sciences 2025;2025(3):375-389
ObjectiveTo explore the pharmacological effects and molecular mechanisms of a Jiao'ai decoction (JAD) in treating premature ovarian failure. Specifically, we evaluated the receptor for advanced glycation end (RAGE) products pathway, metabolic disorders, and intestinal flora dysbiosis.MethodsForty female rats with normal estrous cycles were randomly divided into five groups: control, model, estradiol, low-dose JAD, and high-dose JAD, with 8 rats in each. Except for the control group, the rats in other groups were injected intraperitoneally with cisplatin for 8 days (1.5 mg/kg) to establish a premature ovarian failure model. Starting on the fifth day of cisplatin injections, the estradiol, low-dose JAD, and high-dose JAD groups were administered corresponding drugs for 21 days. Sex hormone levels and pathological changes in the ovaries were measured. Key proteins in the RAGE pathway related to apoptosis, aging, and inflammation, were tested using Western blot. A 16S rRNA analysis of feces and non-targeted metabolism in serum was performed to determine the effects of JAD on intestinal flora and metabolism.ResultsBody weight, ovarian index, and the number of follicles at all levels increased in the JAD group. Regarding serum hormones, estradiol, anti-mullerian hormone (AMH) and P levels increased, whereas follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels decreased in the JAD group. The levels of phosphorylated Akt protein (P-Akt), receptor for advanced glycation end products (RAGE), tumor protein p53 (P53), C-reactive protein (CRP), apoptosis regulator BAX (BAX) and Caspase3 were downregulated by JAD, whereas B cell leukemia/lymphoma 2 (Bcl-2) and Endothelial nitric oxidase synthase (eNOS) were upregulated. JAD was also found to play an important role in the regulation of metabolic disorders and intestinal ecological imbalances by adjusting species composition and diversity.ConclusionJAD can protect ovaries by exerting anti-inflammatory and anti-apoptotic effects via inhibition of the RAGE pathway. JAD can also regulate metabolic disorders and maintain the dynamic balance of intestinal flora, thereby contributing to the improvement of the ovarian reserve function.
7.Adhesion,proliferation,and vascular smooth muscle differentiation of bone marrow mesenchymal stem cells on different electrospinning membranes
Xianjuan SUN ; Qiuhua WANG ; Jinyi ZHANG ; Yangyang YANG ; Wenshuang WANG ; Xiaoqing ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(4):661-669
BACKGROUND:Small diameter artificial vessels are urgently needed to treat coronary artery and peripheral artery diseases in clinical practice.At present,vascular tissue engineering has become the main method for preparing small diameter artificial vessels.Selecting suitable biomaterials and cell sources is the key factor for successful construction of small diameter tissue engineered vessels. OBJECTIVE:To observe the effect of four kinds of electrospinning membrane materials on proliferation,adhesion and differentiation of bone marrow mesenchymal stem cells into vascular smooth muscle cells. METHODS:Bone marrow mesenchymal stem cells were isolated and extracted from SD rats.The bone marrow mesenchymal stem cells were inoculated separately on polycaprolactone(PCL),polycaprolactone-hyaluronic acid(PCL-HA),polycaprolactone-silk-filament proteins(PCL-SF),and polycaprolactone-gelatin(PCL-GEL)electrospinning membrane materials.After 1,3,and 7 days of culture,the cell arrangement on the material was observed under scanning electron microscope.The proliferation and adhesion of the material were observed by phalloidin staining.The mRNA expressions of CD90,Meflin,and transforming growth factor β were detected by qRT-PCR.After 7 days of induced differentiation into vascular smooth muscle cells,the mRNA expression ofɑ-smooth muscle actin on the material was detected by qRT-PCR. RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells were arranged along the fibers of the four kinds of electrospinning membranes under scanning electron microscopy.(2)Phalloidin staining showed the regular distribution of bone marrow mesenchymal stem cells on the four kinds of electrospinning membranes and parallel distribution along the fiber direction.Moreover,PCL-HA,PCL-SF,and PCL-GEL electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells than PCL electrospinning membranes.Compared with PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells.(3)qRT-PCR showed that the four kinds of electrospun membrane materials could maintain the mRNA expression of CD90 and Meflin in bone marrow mesenchymal stem cells,but there was no significant difference between groups(P>0.05).The mRNA expression of transforming growth factor β in PCL-HA,PCL-SF,and PCL-GEL groups was higher than that in PCL group on days 1 and 7(P<0.05),and the mRNA expression of transforming growth factor β in PCL-SF group was higher than that in the other three groups on days 3 and 7(P<0.05).The mRNA expression of transforming growth factor β in PCL-HA group was higher than that in PCL-GEL group on day 7(P<0.05).(4)qRT-PCR showed that the mRNA expression of ɑ-smooth muscle actin in PCL-SF group was higher than that in the other three groups(P<0.05),and that in PCL-HA group was higher than that in PCL group(P<0.05).(5)The results indicate that compared with PCL,PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes combined with bone marrow mesenchymal stem cells are more suitable for the preparation of small diameter tissue engineered vessels.
8.Analysis of Tongue Coating Microbiota Characteristics in Coronary Heart Disease with Qi Deficiency and Blood Stasis Syndrome
Chuhao WANG ; Yongyue LIU ; Zhaoxuan DING ; Xiaoqing ZHANG
Journal of Traditional Chinese Medicine 2025;66(5):501-508
ObjectiveTo explore the characteristics of the tongue coating microbiota in patients of coronary heart disease (CHD) with qi deficiency and blood stasis syndrome. MethodsA total of 27 CHD patients with qi deficiency and blood stasis syndrome, 29 patients with non-qi deficiency and blood stasis syndrome, and 20 healthy individuals were included in this study. The tongue coating microbiota of the participants was analyzed using 16S rDNA high-throughput sequencing technology, followed by Alpha and Beta diversity analyses and comparisons of microbial abundance. ResultsA total of 479 operational taxonomic units (OTUs) were detected, among which 245 OTUs were shared across all three groups. There were 33 OTUs unique to the qi deficiency and blood stasis syndrome group, 21 OTUs unique to the non-qi deficiency and blood stasis syndrome group, and 121 OTUs unique to the healthy group. The observed species count (Sob), total species richness (Chao1), abundance-based coverage estimator (ACE), and Shannon diversity index were significantly lower in the qi deficiency and blood stasis syndrome and non-qi deficiency and blood stasis syndrome groups compared to the healthy group (P<0.05). Principal coordinate analysis (PCoA) of the tongue coating microbiota showed significant differences in distance matrices among the three groups (P<0.05). Compared with the healthy group, the qi deficiency and blood stasis syndrome group exhibited an increased abundance of Actinobacteria, Patescibacteria, Spirochaetes, Verrucomicrobia, Rothia, TM7X, Gemella, and Corynebacterium, while Fusobacteria, Cyanobacteria, Leptotrichia, and Lactobacillus decreased (P<0.05). In the non-qi deficiency and blood stasis syndrome group, Actinobacteria, Verrucomicrobia, Rothia, and Corynebacterium increased, whereas Cyanobacteria and Lactobacillus reduced (P<0.05). When comparing with the non-qi deficiency and blood stasis syndrome group, the qi deficiency and blood stasis syndrome group had a significantly higher abundance of Patescibacteria, Peptostreptococcus, Solobacterium, Filifactor, Moraxella, Porphyromonas endodontalis, and Capnocytophaga, while Cyanobacteria reduced (P<0.05). Conclusuion Patients with CHD of qi deficiency and blood stasis syndrome exhibit a decrease in beneficial bacteria and an increase in pathogenic bacteria. Patescibacteria, Peptostreptococcus, Solobacterium, Filifactor, Moraxella, Porphyromonas endodontalis, and Capnocytophaga were identified as the key differential microbiota distinguishing qi deficiency and blood stasis syndrome from non-qi deficiency and blood stasis syndrome patients.
9.Effect of Zishen Tongguan Formula on "Gut-prostate" Axis of Rats with Chronic Non-bacterial Prostatitis Based on 16S rDNA Sequencing
Xiran LI ; Mengjiao CHEN ; Kaiping ZOU ; Chenguang ZHAO ; Xingbin DAI ; Xiaoqing ZHANG ; Shun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):63-71
ObjectiveBased on the theory of "gut-prostate" axis, this study explored the effects and mechanisms of Zishen Tongguan formula and Cinnamomi Cortex in the formula in treating rats with chronic non-bacterial prostatitis(CNP) by detecting the levels of inflammatory factors, and the composition and structure of intestinal flora in CNP rats. MethodsEight out of 42 SD rats were randomly selected as the normal group, and the remaining rats were injected with carrageenan to prepare the CNP model. After successful modeling, 32 rats were randomly divided into the model group, Ningmitai capsule group(0.50 g·kg-1), Zishen Tongguan formula group(2.00 g·kg-1), and the Phellodendri Chinensis Cortex-Anemarrhenae Rhizoma pair group(PCC-AR group, 2.00 g·kg-1), with 8 rats in each group. The administered groups were given the corresponding medicinal solution by gavage, and the normal and model groups were intragastrically administered with an equal volume of normal saline, once a day for 14 consecutive days. The prostate tissues of rats were collected and subjected to hematoxylin-eosin(HE) staining and Masson staining to observe the pathological changes of the tissues in each group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of related inflammatory factors in rat serum, and 16S rDNA sequencing was used to analyze the abundance and diversity changes of gut microbiota before and after administration, and species difference analysis was performed. ResultsAll the administered groups could alleviate the inflammatory symptoms of CNP rats, increase the expression levels of anti-inflammatory factors and decrease the expression levels of pro-inflammatory factors, with the most sIgnificant effect observed in the Zishen Tongguan formula group. Compared with the normal group, the expression levels of interleukin(IL)-8, hypersensitive C-reactive protein(hs-CRP), immunoglobulin(Ig)M, secretory IgA (sIgA), and inducible nitric oxide synthase(iNOS) were sIgnificantly increased in the model group(P<0.01). Compared with the model group, the expression levels of the above inflammatory factors in all administered groups were significantly reduced(P<0.01). When compared with the PCC-AR group, the Zishen Tongguan formula group showed a significant decrease in transforming growth factor(TGF)-β1 expression level(P<0.05) and a significant increase in IgM expression level(P<0.01). The results of gut microbiota analysis showed that, compared with the PCC-AR group, at the order level, the Zishen Tongguan formula group significantly reduced the relative abundance of conditional pathogens such as Bacteroidales, Acidaminococcales, Rhodospirillales, Clostridiales, and Elusimicrobiales(P<0.01). And at the genus level, the Zishen Tongguan formula group significantly decreased the relative abundance of pathogenic microbiota such as Lachnospira and Bacteroides(P<0.01) and significantly increased the relative abundances of beneficial microbiota such as Ruminococcus and Lactobacillus(P<0.01). ConclusionZishen Tongguan formula can reduce the level of harmful intestinal bacteria, increase the level of beneficial intestinal bacteria, down-regulate the expression of serum inflammatory factors, and the small amount of Cinnamomi Cortex in the formula may play a key role in the treatment of CNP with this formula.
10.Effect of Zishen Tongguan Formula on "Gut-prostate" Axis of Rats with Chronic Non-bacterial Prostatitis Based on 16S rDNA Sequencing
Xiran LI ; Mengjiao CHEN ; Kaiping ZOU ; Chenguang ZHAO ; Xingbin DAI ; Xiaoqing ZHANG ; Shun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):63-71
ObjectiveBased on the theory of "gut-prostate" axis, this study explored the effects and mechanisms of Zishen Tongguan formula and Cinnamomi Cortex in the formula in treating rats with chronic non-bacterial prostatitis(CNP) by detecting the levels of inflammatory factors, and the composition and structure of intestinal flora in CNP rats. MethodsEight out of 42 SD rats were randomly selected as the normal group, and the remaining rats were injected with carrageenan to prepare the CNP model. After successful modeling, 32 rats were randomly divided into the model group, Ningmitai capsule group(0.50 g·kg-1), Zishen Tongguan formula group(2.00 g·kg-1), and the Phellodendri Chinensis Cortex-Anemarrhenae Rhizoma pair group(PCC-AR group, 2.00 g·kg-1), with 8 rats in each group. The administered groups were given the corresponding medicinal solution by gavage, and the normal and model groups were intragastrically administered with an equal volume of normal saline, once a day for 14 consecutive days. The prostate tissues of rats were collected and subjected to hematoxylin-eosin(HE) staining and Masson staining to observe the pathological changes of the tissues in each group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of related inflammatory factors in rat serum, and 16S rDNA sequencing was used to analyze the abundance and diversity changes of gut microbiota before and after administration, and species difference analysis was performed. ResultsAll the administered groups could alleviate the inflammatory symptoms of CNP rats, increase the expression levels of anti-inflammatory factors and decrease the expression levels of pro-inflammatory factors, with the most sIgnificant effect observed in the Zishen Tongguan formula group. Compared with the normal group, the expression levels of interleukin(IL)-8, hypersensitive C-reactive protein(hs-CRP), immunoglobulin(Ig)M, secretory IgA (sIgA), and inducible nitric oxide synthase(iNOS) were sIgnificantly increased in the model group(P<0.01). Compared with the model group, the expression levels of the above inflammatory factors in all administered groups were significantly reduced(P<0.01). When compared with the PCC-AR group, the Zishen Tongguan formula group showed a significant decrease in transforming growth factor(TGF)-β1 expression level(P<0.05) and a significant increase in IgM expression level(P<0.01). The results of gut microbiota analysis showed that, compared with the PCC-AR group, at the order level, the Zishen Tongguan formula group significantly reduced the relative abundance of conditional pathogens such as Bacteroidales, Acidaminococcales, Rhodospirillales, Clostridiales, and Elusimicrobiales(P<0.01). And at the genus level, the Zishen Tongguan formula group significantly decreased the relative abundance of pathogenic microbiota such as Lachnospira and Bacteroides(P<0.01) and significantly increased the relative abundances of beneficial microbiota such as Ruminococcus and Lactobacillus(P<0.01). ConclusionZishen Tongguan formula can reduce the level of harmful intestinal bacteria, increase the level of beneficial intestinal bacteria, down-regulate the expression of serum inflammatory factors, and the small amount of Cinnamomi Cortex in the formula may play a key role in the treatment of CNP with this formula.


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