1.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
2.Proteomics and Network Pharmacology Reveal Mechanism of Xiaoer Huatan Zhike Granules in Treating Allergic Cough
Youqi DU ; Yini XU ; Jiajia LIAO ; Chaowen LONG ; Shidie TAI ; Youwen DU ; Song LI ; Shiquan GAN ; Xiangchun SHEN ; Ling TAO ; Shuying YANG ; Lingyun FU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):69-79
ObjectiveTo explore the pharmacological mechanism involved in the treatment of allergic cough (AC) by Xiaoer Huatan Zhike granules (XEHT) based on proteomics and network pharmacology. MethodsAfter sensitization by intraperitoneal injection of 1 mL suspension containing 2 mg ovalbumin (OVA) and 100 mg aluminum hydroxide, a guinea pig model of allergic cough was constructed by nebulization with 1% OVA. The modeled guinea pigs were randomized into the model, low-, medium- and high-dose (1, 5, 20 g·kg-1, respectively) XEHT, and sodium montelukast (1 mg·kg-1) groups (n=6), and another 6 guinea pigs were selected as the blank group. The guinea pigs in drug administration groups were administrated with the corresponding drugs by gavage, and those in the blank and model groups received the same volume of normal saline by gavage, 1 time·d-1. After 10 consecutive days of drug administration, the guinea pigs were stimulated by 1% OVA nebulization, and the coughs were observed. The pathological changes in the lung tissue were observed by hematoxylin-eosin staining. The enzyme-linked immunosorbent assay was performed to measure the levels of C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), and malondialdehyde (MDA) in the bronchoalveolar lavage fluid (BALF) and immunoglobulin G (IgG) and immunoglobulin A (IgA) in the serum. Immunohistochemistry (IHC) was employed to observe the expression of IL-6 and TNF-α in the lung tissue. Transmission electron microscopy was employed observe the alveolar type Ⅱ epithelial cell ultrastructure. Real-time PCR was employed to determine the mRNA levels of IL-6, interleukin-1β (IL-1β), and TNF-α in the lung tissue. Label-free proteomics was used to detect the differential proteins among groups. Network pharmacology was used to predict the targets of XEHT in treating AC. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to search for the same pathways from the results of proteomics and network pharmacology. ResultsCompared with the blank group, the model group showed increased coughs (P<0.01), elevated levels of CRP, TNF-α, IL-6, and MDA and lowered level of SOD in the BALF (P<0.05, P<0.01), elevated levels of IgA and IgG in the serum (P<0.05, P<0.01), congestion of the lung tissue and infiltration of inflammatory cells, increased expression of IL-6 and TNF-α (P<0.01), large areas of low electron density edema in type Ⅱ epithelial cells, obvious swelling and vacuolization of the organelles, karyopyknosis or sparse and dissolved chromatin, and up-regulated mRNA levels of IL-6, IL-1β, and TNF-α (P<0.01). Compared with the model group, the drug administration groups showed reduced coughs (P<0.01), lowered levels of CRP, TNF-α, IL-6, and MDA and elevated level of SOD in the BALF (P<0.05, P<0.01), alleviated lung tissue congestion, inflammatory cell infiltration, and type Ⅱ epithelial cell injury, and decreased expression of IL-6 and TNF-α (P<0.01). In addition, the medium-dose XEHT group and the montelukast sodium group showcased lowered serum levels of IgA and IgG (P<0.05, P<0.01). The medium- and high-dose XEHT groups and the montelukast sodium showed down-regulated mRNA levels of IL-6, IL-1β, and TNF-α and the low-dose XEHT group showed down-regulated mRNA levels of IL-6 and TNF-α (P<0.05, P<0.01). Phospholipase D, mammalian target of rapamycin (mTOR), and epidermal growth factor receptor family of receptor tyrosine kinase (ErbB) signaling pathways were the common pathways predicted by both proteomics and network pharmacology. ConclusionProteomics combined with network pharmacology reveal that XEHT can ameliorate AC by regulating the phospholipase D, mTOR, and ErbB signaling pathways.
3.Clinical Characteristics and Influencing Factors of Rheumatoid Arthritis in Patients with Cold Dampness Obstruction Syndrome
Yanyu CHEN ; Yanqi LI ; Longxiao LIU ; Liubo ZHANG ; Tianyi LAN ; Nan ZHANG ; Cheng XIAO ; Yuan XU ; Qingwen TAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):140-146
ObjectiveTo study the clinical characteristics and influencing factors of rheumatoid arthritis (RA) in the patients with cold dampness obstruction syndrome. MethodsThe RA patients treated in the Department of Traditional Chinese Medicine and Rheumatology of the China-Japan Friendship Hospital from August 2022 to June 2024 were selected. The demographic information, clinical data, laboratory test results, and traditional Chinese medicine (TCM) symptom information were collected for syndrome differentiation, on the basis of which the characteristics and influencing factors of cold dampness obstruction syndrome were analyzed. ResultsA total of 258 RA patients were selected in this study, including 88 (34.1%) patients with cold dampness obstruction syndrome, 53 (20.5%) patients with dampness and heat obstruction syndrome, 31 (12.0%) patients with wind dampness obstruction syndrome, 29 (11.2%) patients with liver-kidney deficiency syndrome, 19 (7.4%) patients with Qi-blood deficiency syndrome, 14 (5.4%) patients with phlegm-stasis obstruction syndrome, 15 (5.8%) patients with stasis obstructing collateral syndrome and 9 (3.5%) patients with Qi-Yin deficiency syndrome. The patients were assigned into two groups of cold dampness obstruction syndrome and other syndromes. The group of cold dampness obstruction syndrome had lower joint fever, 28-tender joint count (TJC28), and 28-joint disease activity score (DAS28)-C-reactive protein (CRP) and higher central sensitization, cold feeling of joints, fear of wind and cold, cold limbs, and abdominal distention than the group of other syndromes (P<0.05). The binary logistic regression analysis showed that central sensitization (OR 5.749, 95%CI 2.116-15.616, P<0.001) and DAS28-CRP (OR 0.600, 95% CI 0.418-0.862, P=0.006) were the independent factors influencing cold dampness obstruction syndrome in RA. ConclusionCold dampness obstruction syndrome is a common syndrome in RA patients. It is associated with central sensitization, cold feeling of joints, abdominal distension and may be a clinical syndrome associated with central sensitization.
4.Analysis of Quality Uniformity of Hengzhi Kechuan Capsules Based on HPLC-DAD-CAD
Qian MA ; An LIU ; Qingxia XU ; Cong GUO ; Jun ZHANG ; Maoqing WANG ; Xiaodi KOU ; Yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):168-174
ObjectiveTo establish the fingerprints of 15 batches of Hengzhi Kechuan capsules, to quantitatively analyze 10 index components, and to evaluate the quality uniformity of samples from different batches. MethodsThe fingerprints and quantitative analysis of Hengzhi Kechuan capsules were established by a combination method of high performance liquid chromatography coupled with diode array detector and charged aerosol detector(HPLC-DAD-CAD), adenosine, guanosine, vanillic acid, safflomin A, agarotetrol, naringin, hesperidin, militarine, ginsenoside Rb1, and glycyrrhizic acid were selected as quality attribute indexes. A total of 15 batches of Hengzhi Kechuan capsules from 2022 to 2024(3 boxes per batch) were qualitatively and quantitatively analyzed, and the quality uniformity level of the manufacturers was characterized by parameters of intra-batch consistency(PA) and inter-batch consistency(PB). The homogeneity and difference of quality attribute indexes of samples from different years were analyzed by heatmap clustering analysis. ResultsHPLC fingerprints and quantitative method of Hengzhi Kechuan capsules were established, and the methods could be used for qualitative and quantitative analysis of this preparation, which was found to be stable and reliable by method validation. The similarity of fingerprints of 15 batches of samples was 0.887-0.975, a total of 13 common peaks were calibrated, and 10 common peaks were designated, all of which were quality attribute index components. The results of quantitative analysis showed that the contents of the above 10 ingredients in the samples were 0.038-0.078, 0.115-0.251, 0.007-0.018, 0.291-0.673, 0.122-0.257, 0.887-1.905, 1.841-3.364, 1.412-2.450, 2.207-3.112, 0.650-1.161, respectively. And the contents of ginsenoside Rb1 and glycyrrhizic acid met the limit requirements in the 2020 edition of Chinese Pharmacopoeia. For the samples from 15 batches, the PA values of the 10 index components were all <10%, indicating good intra-batch homogeneity, and the PB values ranged from 33.86% to 92.97%, suggesting that the inter-batch homogeneity was poor. Heatmap clustering analysis showed that the samples from different years were clustered into separate categories, and adenosine, guanosine, safflomin A, naringin, hesperidin and agarotetrol were the main differential components. ConclusionThe intra-annual quality uniformity of Hengzhi Kechuan capsules is good and the inter-annual quality uniformity is insufficient, which may be related to the quality difference of Pinellinae Rhizoma Praeparatum, Carthami Flos, Citri Sarcodactylis Fructus, Citri Reticulatae Pericarpium, Aquilariae Lignum Resinatum, Citri Fructus, etc. In this study, the fingerprint and multi-indicator determination method of Hengzhi Kechuan capsules was established, which can be used for more accurate and efficient quality control and standardization enhancement.
5.ZHANG Lei's Experience in the Treatment of Sjögren Syndrome from the Perspective of "Same Form and Disease of Dryness and Dampness"
Yanan LI ; Erping XU ; Rui ZHANG
Journal of Traditional Chinese Medicine 2025;66(1):12-16
This paper summarized professor ZHANG Lei's experience in the treatment of sjögren syndrome (SS) from the perspective of "same form and disease of dryness and dampness". It is believed that both dryness and dampness are involved as the pathological factors of SS. The important pathogenesis is that dryness and dampness are of the same form and coexist in the disease, that is, dryness and dampness can transform into each other with the same form or be concurrent in the disease. The same form of dryness and dampness includes the dryness syndrome caused by dampness and the dampness syndrome caused by dryness, which can be treated with modified Wuling Powder (五苓散) and modified Ejiao Jizihuang Decoction (阿胶鸡子黄汤), respectively. The disease of both dryness and dampness includes lung dryness with spleen dampness, stomach dryness with spleen dampness, liver dryness with spleen dampness, and kidney dryness with spleen dampness syndrome, which can be treated with modified Ganlu Beverage and Weijing Decoction(甘露饮合苇茎汤), self-made Jianpi Yangyin Formula (健脾养阴方) with modifications, self-made Guqing Decoction and Jupi Zhishu Pill (谷青汤合橘皮枳术丸) with modifications, and Shenqi Pill and Linggui Zhugan Decoction (肾气丸合苓桂术甘汤) with modifications, respectively.
6.Development of Patient Self-Reported Core Outcome Set in Community Studies on Heat-Sensitive Moxibustion for Primary Hypertension
Jianyu YOU ; Shuqing LI ; Guihua DENG ; Xu ZHOU
Journal of Traditional Chinese Medicine 2025;66(1):34-41
ObjectiveTo establish the self-reported core outcome set (COS) for patients with an example of a community study on heat-sensitive moxibustion for primary hypertension (PH), to provide a reference for the selection of effectiveness evaluation indicators in community study on heat-sensitive moxibustion. MethodsA systematic literature search was conducted to collect outcomes used in randomized controlled trials and systematic review of heat-sensitive moxibustion for PH (Jan 2021), and additional outcomes were added through patient and expert questionnaires (Feb 2021) to create a pool of outcome entries. A multidisciplinary expert Delphi survey was conducted to screen outcomes applicable to patient self-reporting (Apr 2021), and the importance of outcome indicators was rated on a 5-Point Likert Scale. Finally, patient self-reported COS was determined through a consensus conference (June 2021). ResultsA pool of patient self-reported indicators in the community study of heat-sensitive moxibustion treatment for PH was generated by standardizing and combining the outcome indicators based on the results of the literature search and the questionnaire survey, which consisted of totally 100 measurement tools or contents, excluding 51 indicators or measurement tools required measurement by specialized physicians or hospital equipment, and 49 items were retained to enter the initial list of indicator entries. For the first round of Delphi survey, the mean score for expert familiarity was 0.819, the mean score for basis of judgment was 0.710, and the expert authority coefficient was 0.765, with a total of 21 indicator measurement tools or contents deleted (significance score ≤ 75 or coefficient of variation > 0.25), 28 retained, and 3 new expert-added indicator entries added. In the second round of Delphi survey, the average score for expert familiarity was 0.859, the average score for basis for judgment was 0.763, and the expert authority coefficient was 0.811, with a total of 11 indicator measurement tools or contents deleted and 20 retained involving 5 domains. Following an expert consensus meeting, 8 outcome indicators were finalized for inclusion in the patient self-reported COS, including 6 indicators of effectiveness evaluation such as quality-of-life scores, blood pressure, traditional Chinese medicine symptom scores, cost-benefit, cardiovascular and cerebrovascular events, and adverse reactions/events, and 2 indicators of factors influencing effectiveness such as sensation of heat-sensitive moxibustion, and adherence. ConclusionIn this study, we initially established a criteria for evaluating the effectiveness in the community study on heat-sensitive moxibustion by constructing patient self-reported COS in the community study on heat-sensitive moxibustion for PH, which can provide a scientific research paradigm for the subsequent development of the community study on heat-sensitive moxibustion.
7.Effects of Shenfuhuang Formula (参附黄配方) on Potential Targets of Action in the Brain Tissue of Sepsis Model Mice:Transcriptomics-Based Exploration
Yuchen WANG ; Xuerui WANG ; Xiaolong XU ; Jingxia ZHAO ; Jiabo WANG ; Yuan GAO ; Weijun KONG ; Qingquan LIU
Journal of Traditional Chinese Medicine 2025;66(1):65-70
ObjectiveTo investigate the possible mechanism of Shenfuhuang Formula (参附黄配方) in prevention and treatment of epsis-associated encephalopathy from the perspective of brain genomics. MethodsC57BL/6 mice were randomly divided into sham surgery group, sepsis group, and Shenfuhuang group, with 20 mice in each group. The sepsis group and Shenfuhuang group were induced to develop sepsis by cecal ligation and puncture (CLP) procedure. At 4 hours after modelling, Shenfuhuang group were gavaged with 2.5 g/(kg·d) of Shenfuhuang Formula, 0.5 ml each time, at 12 hours intervals, for a total of 4 times after modelling. Sepsis group and sham surgery group were given 0.5 ml of purified water orally. At 48 hours after modeling, the transcriptome sequencing was used to explore the differential gene expression in the effects of Shenfuhuang Formula on the brain regions of septic mice, and real-time PCR and ELISA were later used to further validate the differential gene and proteins expression. ResultsA total of 4605 genes were differentially expressed in Shenfuhuang group compared with sepsis group, of which 2353 genes were up-regulated and 2252 genes were down-regulated. According to the results of previous publications, six key genes were screened, including serine/threonine-protein kinase (Nek1), myelin-associated glycoprotein (Mag), endothelial cell-specific tyrosine kinase receptor (Tek), a disintegrin and metalloproteinase with thrombospondin motifs 20 (Adamts20), lymphocyte antigen 86 (Ly86), and E3 ubiquitin-protein ligase (Traip). Further genetic and protein validation revealed that, compared to the sham surgery group, the mRNA levels and corresponding protein levels of Nek1, Mag, Tek, Adamts20, Ly86, and Traip in the brain tissue of septic mice significantly reduced (P<0.05). In comparison to the sepsis group, Shenfuhuang group showed significantly increased mRNA levels and corresponding protein levels of Nek1, Mag, Tek, Adamts20, Ly86, and Traip (P<0.05). ConclusionThe potential therapeutic targets of Shenfuhuang Formula for treating sepsis-associated encephalopathy may be related to the Nek1, Mag, Tek, Adamts20, Ly86, and Traip genes and their encoded proteins.
8.The Effect of Modified Ditan Decoction (涤痰汤) on Cognitive Function and Resting-State Functional Magnetic Resonance Imaging of the Brain in Chronic Intermittent Hypoxia Model Rats
Naijie CHEN ; Xiaoting WANG ; Fengsheng XU ; Shuanghong SHEN ; Zuanfang LI ; Qin CHEN ; Jin CHEN ; Runhua WU
Journal of Traditional Chinese Medicine 2025;66(1):71-78
ObjectiveTo explore the effect of modified Ditan Decoction (涤痰汤) on chronic intermittent hypoxia cognitive function and the potential function mechanism. MethodsTwenty-four Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, and a modified Ditan Decoction group, with eight rats in each group. Rats in the modified Ditan Decoction group were administered the decoction by gavage at 14.8 ml/(kg·d), while the normal group and the model group received the same dose of normal saline. Thirty minutes after daily gavage, the rats in all three groups were placed in an intermittent hypoxia chamber. The oxygen concentration for the model group and the modified Ditan Decoction group was adjusted daily for 8 hours using a computer program to establish the model, while the normal group was exposed to the same airflow rate of ambient air. The intervention was continued for 12 weeks to establish a chronic intermittent hypoxia rat model. The Y-maze test was used to evaluate spatial working memory in the rats. Resting-state functional magnetic resonance imaging (rs-fMRI) was performed to detect whole-brain regional homogeneity (ReHo) and seed-based functional connectivity (FC). Brain regions showing significant differences in rs-fMRI were selected for further analysis. Immunofluorescence was used to detect β-amyloid (Aβ) deposition and the number of ionized calcium-binding adapter molecule 1 (IBA1)-positive microglial cells. Immunohistochemistry was employed to assess the expression of synaptophysin (SYP), the excitatory synapse marker vesicular glutamate transporter 1 (Vglut1), and the inhibitory synapse marker vesicular γ-aminobutyric acid transporter (VGAT). ResultsCompared with the normal group, the model group showed a reduced spontaneous alternation rate in the Y-maze test. The smoothed Z-score standardized regional homogeneity (SzReHo) value in the left entorhinal cortex significantly increased, and the FC value from this seed point to the left basal forebrain significantly reduced. Additionally, the model group exhibited significantly higher Aβ fluorescence intensity and Iba1 positivity in the left entorhinal cortex, decreased expression of SYP, Vglut1, and VGAT, along with an increased Vglut1/VGAT ratio (P<0.05 or P<0.01). Compared to the model group, the modified Ditan Decoction group demonstrated an increased spontaneous alternation rate, a significantly reduced SzReHo value in the left entorhinal cortex, and a significantly increased FC value from this region to the left basal forebrain. Furthermore, this group showed significantly lower Aβ fluorescence intensity and Iba1 positivity in the left entorhinal cortex, increased levels of SYP, Vglut1, and VGAT, and a decreased Vglut1/VGAT ratio (P<0.05 or P<0.01). ConclusionModified Ditan Decoction can reconstruct the projection from the left basal forebrain to the entorhinal cortex in chronic intermittent hypoxia, thereby reducing Aβ aggregation and excessive microglial activation in the left entorhinal cortex. This process improves the excitation/inhibition imbalance caused by synaptic remodeling, ultimately enhancing cognitive function in rats of chronic intermittent hypoxia.
9.The Effect of Qishao Tongbi Capsule (芪芍通痹胶囊) on the Wnt/β-catenin Pathway in a Rat Model of Intervertebral Disc Degeneration
Yumen XUE ; Xilin XU ; Wei HAN ; Jiaben XU ; Wenting XU ; Zelin LIU ; Xiaofeng ZHANG
Journal of Traditional Chinese Medicine 2025;66(1):79-88
ObjectiveTo explore the possible mechanism of Qishao Tongbi Capsule (芪芍通痹胶囊, QTC) in the treatment of intervertebral disc degeneration (IDD). MethodsSeventy-five rats were randomly divided into control group, model group, low-dose QTC group, high-dose QTC group, high-dose QTC +agonist group, with 15 rats in each group. Except for the control group, all other groups were subjected to a fibrous ring puncture to prepare an IDD model. After modeling, rats in low-dose QTC group and high-dose QTC group were given QTC at doses of 0.2 and 0.8 g/(kg·d) by gavage, respectively. Rats in high-dose QTC+ agonist group was given QTC at 0.8 g/(kg·d) and SKL2001 solution at 10 mg/(kg·d) by gavage. The control group and model group were given 10 ml/(kg·d) distilled water by gavage. All treatments were given once a day for 4 consecutive weeks. After treatment, X-ray and magnetic resonance imaging (MRI) were used to detect IDD degree. Hematoxylin-eosin (HE) staining and Safranin O-Fast Green staining were used to observe the morphological changes of the intervertebral disc tissue. Immunohistochemical staining was performed to examine the levels of proteoglycan, type Ⅱ collagen (COL Ⅱ), and matrix metalloproteinase-3 (MMP-3) in the intervertebral disc tissue. Western blotting was used to detect the extracellular matrix (ECM)-related proteins (proteoglycan, COL Ⅱ, MMP-3, MMP-9, MMP-13), aging-related proteins (P53, P21, P16), apoptosis related proteins, including B-cell lymphoma/leukemia 2 (BCL-2), BCL-2 related X protein (BAX), Cleaved Caspase-3, and Wnt/β-catenin pathway related proteins such as Wnt3a, glycogen synthase kinase-3β (GSK-3β) and β-catenin in the intervertebral disc nucleus pulposus (NP) tissue. Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) was used to assess the mRNA expression of Wnt3a, GSK-3β, and β-catenin in intervertebral disc tissue. ResultsCompared with the model group, rats in the low-dose QTC group and high-dose QTC group exhibited improved DHI, decreased Pfirmann grading, and alleviated IDD. The structural integrity of the NP and annulus fibrosus increased, and the number of the NP increased. The levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β increased, while the levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX, Cleaved Caspase-3, Wnt3a and β-catenin protein decreased. The mRNA expression of Wnt3a and β-catenin mRNA decreased, while GSK-3β mRNA expression increased (P<0.05). Compared with the low-dose QTC group, the high-dose QTC group showed further improvements in DHI, decrease in Pfirrmann grading (P<0.05), and greater alleviation of IDD. The structural integrity of NP and annulus fibrosus was further enhanced, and the number of NP cells further increased. The levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β were higher, while the levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX, Cleaved Caspase-3, Wnt3a and β-catenin were lower. The mRNA expression of Wnt3a and β-catenin decreased, while GSK-3β mRNA expression increased (P<0.05). Compared with the high-dose QTC group, the high-dose QTC +agonist group showed a decrease of DHI, an increase of Pfirmann grading (P<0.05), significant aggravation of IDD, reduction in structural integrity of the NP and annulus fibrosus, a decrease of NP cell count, lower levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β, and higher levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX and Cleaved Caspase-3. Additionally, GSK-3β mRNA expression decreased (P<0.05). ConclusionQTC can inhibit NP cell aging, apoptosis, and ECM degradation in IDD rats, and its therapeutic effect may be mediated through the inhibition of the Wnt/β-catenin pathway.
10.Survey on the perception and current status of drug risk management in medical institutions
Xuelin SUN ; Mingqing XING ; Zixuan ZHANG ; Wenjing ZHAO ; Dongfang QIAN ; Yan LIANG ; Li XU ; Pengfei JIN ; Yatong ZHANG
China Pharmacy 2025;36(1):7-12
OBJECTIVE To know about the perception and current status of drug risk management among pharmacists in Chinese medical institutions, providing insights and recommendations for enhancing the drug risk management system in medical institutions. METHODS A questionnaire survey was conducted across 28 provinces, cities, and autonomous regions; stratified radom sampling was employed to study the population of medical workers and pharmaceutical professionals in medical institutions nationwide. The survey included information on the survey population, the current status of drug risk management implementation in medical institutions, the cognition, definition and process of drug risk management related concepts, and the content and mode of drug risk management work in medical institutions. Finally, suggestions were collected from various medical institutions on the system construction of drug risk management. Descriptive statistical analysis was adopted to summarize the obtained data. RESULTS A total of 446 questionnaires were collected in this survey, including 420 valid questionnaires and 26 invalid questionnaires. The questionnaire collection rate was 100%,and the effective rate was 94.17%. 51.19% of the respondents No.2020YFC2009001)。 based their understanding of drug risk management on Management Measures for Adverse Drug Reaction Reports and Monitoring, while 87.38% recognized the need for drug risk management throughout the drug use process. 63.33% of the participants stated that their medical institutions had dedicated positions related to drug risk management, with the highest proportion (72.17%) was in third-grade class A medical institutions. 66.43% reported implementing risk management across all drug use stages. Suggestions for the development of drug risk management systems in medical institutions by the research participants focused on enhancing guiding documents, clarifying concepts, establishing information-sharing mechanisms. CONCLUSIONS The overall awareness of drug risk management in China’s medical institutions is high, with practices in place across various stages in multiple forms. However, there remains a need to strengthen institutional documents, management regulations, system development, and information-sharing mechanisms to improve collaborative governance, improve drug management levels, and ensure patient safety.

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