1.Nuangong Tiaojing Decoction (暖宫调经汤) Combined with Ginger Moxibustion at Baliao (八髎) Acupoints for Patients with Premature Ovarian Failure of Yang-Deficiency Uterus-Coldeness Syndrome:65 Cases Clinical Observation
Yuqiong MENG ; Qiang GAO ; Tingting ZHAI ; Zhihong YANG
Journal of Traditional Chinese Medicine 2025;66(1):59-64
ObjectiveTo explore the clinical effectivess of Nuangong Tiaojing Decoction (暖宫调经汤) combined with ginger moxibustion at Baliao acupoints (Eight Bone-Holes) for treating premature ovarian failure (POF) of yang-deficiency uterus-coldeness syndrome, based on conventional western medical treatment. MethodsA total of 130 patients diagnosed with POF of yang-deficiency uterus-coldeness syndrome were divided into a treatment group and a control group, with 65 cases in each group, based on patient preference. The control group received conventional western medicine treatment, while the treatment group was additionally given Nuangong Tiaojing Decoction orally one dose per day and underwent ginger moxibustion at Baliao points once daily. Both groups were treated for three months. Before and after treatment, ovarian function indicators, including ovarian volume, follicle count, ovarian peak systolic velocity (PSV), and endometrial thickness were assessed, and serum sex hormone levels including progesterone (P), estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and testosterone (T), as well as anti-Müllerian hormone (AMH) levels were also measured. Additionally, traditional Chinese medicine (TCM) syndrome scores were evaluated, covering symptoms such as reduced libido, lumbosacral pain, fear of cold with cold extremities, fatigue, dull complexion, lower abdominal distension and pain, pale tongue, and deep and rough pulse. After treatment, clinical effectiveness and safety were determined. ResultsCompared with the pre-treatment levels within group, both groups showed significant increases in ovarian volume, follicle count, PSV, endometrial thickness, and levels of P, E2, and AMH after treatment, while the levels of FSH, LH, PRL, and T, as well as scores of lumbosacral pain, reduced libido, dull complexion, deep and rough pulse, and total TCM syndrome scores significantly reduced after treatment. Furthermore, the treatment group exhibited higher ovarian volume, follicle count, PSV, endometrial thickness, and levels of P, E2, and AMH compared to the control group, while FSH, LH, PRL, T levels, and all symptom scores, as well as total TCM syndrome scores, were significantly lower in the treatment group than in the control group (P<0.05 or P<0.01). The clinical effectiveness in treatment group was 96.92% (63/65), significantly higher than 81.54% (53/65) in control group (P<0.05). The incidence of adverse reactions in the treatment group was 26.15% (17/65), while in the control group it was 32.31% (21/65), with no statistically significant difference (P>0.05). ConclusionOn the basis of conventional western medicine treament, Nuangong Tiaojing Decoction combined with ginger moxibustion at Baliao acupoints for patients with POF of yang-deficiency uterus-coldeness syndrome can significantly improve ovarian function and clinical symptoms, regulate hormone levels, and thereby enhance clinical effectiveness.
2.Mechanism of Xiezhuo Jiedu Formula in Treating Ulcerative Colitis Through Pyroptosis Regulation Based on Bioinformatics and Animal Experiments
Qiang CHUAI ; Wenjing ZHAI ; Shijie REN ; Xiaomeng LANG ; Xin KANG ; Wenli WEI ; Jingyuan LIU ; Jianping LIU ; Jie REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):105-113
ObjectiveThis study aims to explore the potential mechanism of the Xiezhuo Jiedu formula in regulating pyroptosis for the treatment of ulcerative colitis (UC) using bioinformatics and in vivo animal experiments. MethodsDifferentially expressed genes (DEGs) in colon tissues of UC patients were retrieved from the Gene Expression Omnibus (GEO) database. Pyroptosis-related genes were obtained from the GEO and GeneCards databases. The intersection of these datasets yielded pyroptosis-related DEGs (Pyro-DEGs). Pyro-DEGs were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis using the Metascape database. A protein-protein interaction (PPI) network was constructed using the STRING database. Least absolute shrinkage and selection operator (LASSO) prediction model and receiver operating characteristic (ROC) analysis were conducted to identify core Pyro-DEGs with diagnostic and therapeutic potential. Immune infiltration analysis of the UC datasets was performed using the deconvolution method (CIBERSORT), along with correlation analysis with core Pyro-DEGs. Sixty male Sprague-Dawley (SD) rats were randomly divided into a control group, a model group, high-, medium-, and low-dose groups of Xiezhuo Jiedu formula (26.64, 13.32, 6.66 g·kg-1), and a mesalazine group (0.27 g·kg-1), with 10 rats in each group. UC was established by intrarectal administration of 3,5-trinitrobenzenesulfonic acid (TNBS) dissolved in ethanol. The control and model groups were given distilled water by gavage, while the treatment groups were administered the corresponding drugs for 7 consecutive days. Hematoxylin-eosin (HE) staining was used to observe the colon histopathology. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors such as interleukin-1β (IL-1β), IL-10, IL-18, and transforming growth factor-β (TGF-β). Immunohistochemistry (IHC) and Western blot were applied to detect the expression of Caspase-1, gap junction alpha-1 protein (GJA1), peroxisome proliferator-activated receptor gamma (PPARG), and S100 calcium-binding protein A8 (S100A8). Real-time quantitative polymerase chain reaction (Real-time PCR) was utilized to measure mRNA expression of Caspase-1, GJA1, PPARG, and S100A8. Western blot was performed to assess protein expression levels of Caspase-1, GJA1, PPARG, and S100A8. ResultsGEO datasets GSE87466 and GSE87473 yielded 64 Pyro-DEGs. KEGG analysis indicated that these genes were enriched in the NOD-like receptor signaling pathway, tumor necrosis factor (TNF) signaling pathway, and hypoxia-inducible factor 1 (HIF-1) signaling pathway. Four core Pyro-DEGs (Caspase-1, GJA1, PPARG, and S100A8) were identified. Immune infiltration analysis showed that expression of these genes was positively correlated with mast cells, neutrophils, M0 macrophages, M1 macrophages, and dendritic cells. Animal experimental results indicated that compared with the control group, the model group had significantly increased levels of IL-1β and IL-18, significantly decreased levels of IL-10 and TGF-β. The model group showed enhanced Caspase-1, GJA1, and S100A8 staining, and significantly increased mRNA and protein expression of Caspase-1, GJA1, and S100A8 (P<0.01). In contrast, the expression of PPARG was reduced in the model group (P<0.01). After treatment, all dosage groups showed varying degrees of improvement (P<0.05, P<0.01), with the high-dose group showing the most significant improvement (P<0.01). ConclusionCaspase-1, GJA1, PPARG, and S100A8 are core Pyro-DEGs closely associated with the pathogenesis of UC. These genes may collaborate with immune cells such as mast cells, neutrophils, and M0 macrophages to mediate disease development. The Xiezhuo Jiedu formula may regulate the expression of core Pyro-DEGs through the NOD-like receptor, TNF, and HIF-1 core signaling pathways, thereby modulating immune homeostasis in UC rats and effectively alleviating UC.
3.Research progress on endogenous small-molecule phenolics and the proposal of "phenolomics"
Hong-qian KUI ; Chuan-xin LIU ; Qiang WANG ; Hai-feng ZHAI ; Jian-mei HUANG
Acta Pharmaceutica Sinica 2024;59(2):336-349
Small-molecule phenolic substances widely exist in animals and plants, and have some shared biological activities. The metabolism of phenylalanine and tyrosine in the human body, and especially the metabolism of catecholamine neurotransmitters, produces endogenous small-molecule phenols. Endogenous small-molecule phenolic substances are functionally related to the important physiological processes and the occurrence of mental diseases in humans and some animals, which are systematically sorts and summarized in this review. Integrating the previous experimental research and literature analysis on natural small-molecule phenols by our research group, the understanding of the hypothesis that "small-molecule phenol are pharmacological signal carriers" was deepened. Based on above, the concept of "phenolomics" was further proposed, analyzed the research direction and research content which can bring into the knowledge framework of phenolomics. The induction of phenolomics will provide wider perspectives on explaining the pharmacological mechanism of drugs, discovering new drug targets, and finding biomarkers of mental diseases.
4.Improvement of Colonic Mucosa Inflammatory Response in Mice with Ulcerative Colitis by Xiezhuo Jiedu Recipe Through miRNA-155-5p/JAK2/STAT3 Pathway
Chaodi SUN ; Mengmeng ZHAO ; Xiaomeng LANG ; Jie REN ; Xin KANG ; Jiancong CUI ; Sujie JIA ; Yujing MA ; Yue LIU ; Qiang CHUAI ; Wenjing ZHAI ; Jianping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):174-182
ObjectiveThe differential expression of microRNAs (miRNAs) between the active stage and the remission stage of ulcerative colitis (UC) was analyzed by bioinformatics method, and the regulatory relationship was constructed by screening the differentially expressed genes (DEGs). The mechanism of Xizhuo Jiedu recipe in the treatment of UC was speculated and verified by animal experiments. MethodThe miRNAs data set of colonic mucosa tissue of UC patients was obtained from the gene expression database (GEO), and the most differentially expressed miRNAs were screened by GEO2R, Excel, and other tools as research objects. TargetScan, miRTarbase, miRDB, STRING, TRRUST, and Matescape databases were used to screen key DEGs, predict downstream transcription factors (TFs), gene ontology (GO), and conduct Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key signaling pathways were selected for animal experiments. In animal experiments, the UC mouse model was prepared by making the mouse freely drink 2.5% dextran sodium sulfate (DSS). Xiezhu Jiedu recipe and mesalazine were given by gavage for seven days, and the inflammatory infiltration of colonic mucosa was observed by hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-155-5p in colon tissue. Immunohistochemistry and Western blot were used to detect the protein expression levels of cytokine signal transduction inhibitor (SOCS1), phosphorylated transcriptional signal transductor and activator 3 (p-STAT3), phosphorylated Janus kinase 2 (p-JAK2), and retinoic acid-associated orphan receptor-γt (ROR-γt). The expression levels of transforming growth factor-β (TGF-β), interleukin-17 (IL-17), interleukin-6 (IL-6), and interleukin-10 (IL-10) in serum were detected by enzyme linked immunosorbent assay (ELISA). ResultThe GSE48957 dataset was screened from the GEO database, and miR-155-5p was selected as the research object from the samples in the active and remission stages. 131 DEGs were screened. The GO/KEGG enrichment analysis was closely related to biological processes such as positive regulation of miRNA transcription and protein phosphorylation, as well as signaling pathways such as stem cell signaling pathway, IL-17 signaling pathway, and helper T cell 17 (Th17) cell differentiation. The Matescape database was used to screen out 10 key DEGs, among which SOCS1 was one of the key DEGs of miR-155-5p. Further screening of the TFS of key DEGs revealed that STAT3 was one of the main TFs of SOCS1. The results of animal experiments showed that Xiezhu Jiedu Recipe could effectively down-regulate the mRNA expression of miR-155-5p and protein expression of p-STAT3, p-JAK2, and ROR-γt in colon tissue of UC mice and the expression of IL-17 and IL-6 in serum of UC mice, up-regulate the protein expression of SOCS1 and the expression of TGF-β and IL-10, increase the level of anti-inflammatory factors, and reduce inflammatory cell infiltration. ConclusionIt is speculated that Xizhuo Jiedu recipe may interfere with SOCS1 by regulating the expression of miR-155-5p in UC mice, inhibit the phosphorylation of STAT3, inhibit the differentiation of CD4+ T cells into Th17 cells, reduce the levels of pro-inflammatory factors (IL-17 and IL-6), and increase the levels of anti-inflammatory factors (TGF-β and IL-10). As a result, the inflammation of colon mucosa in UC mice was alleviated.
5.Value of noninvasive echocardiographic indicators in predicting pulmonary vascular resistance in chronic thromboembolic pulmonary hypertension
Yanan ZHAI ; Aili LI ; Wanmu XIE ; Qiang HUANG ; Qian GAO ; Yu ZHANG ; Aihong CHEN ; Guangjie LYU ; Jieping LEI ; Zhenguo ZHAI
Chinese Journal of Ultrasonography 2024;33(2):134-141
Objective:To investigate the values of two-dimensional and three-dimensional echocardiographic parameters in predicting pulmonary vascular resistance (PVR) in chronic pulmonary thromboembolic pulmonary hypertension (CTEPH).Methods:A total of 141 patients diagnosed with CTEPH in China-Japan Friendship Hospital from November 2015 to December 2022 were included. Two-dimensional echocardiographic indicators reflecting PVR were constructed according to the calculation formula of PVR: echocardiographic estimated systolic pulmonary artery pressure (sPAP Echo)/left ventricular end-diastolic diameter (LVIDd), echocardiographic estimated mean pulmonary artery pressure (mPAP Echo)/LVIDd. sPAP Echo/left ventricular end-diastolic volume (LVEDV), sPAP Echo/left ventricular cardiac output (LVCO) were measured by three-dimensional echocardiography. The correlations between two-dimensional and three-dimensional echocardiographic ratios and invasive PVR were then analyzed using the Spearman correlation method. Using receiver operating characteristic curve analysis, cut-off values for the ratios were generated to identify patients with PVR>1 000 dyn·s -1·cm -5. Pre- and postoperative hemodynamics and echocardiographic data were analyzed, as well as the correlation between the reduction rate of the echocardiographic index and PVR in 54 patients who underwent pulmonary endarterectomy (PEA). Results:sPAP Echo/LVIDd, sPAP Echo/LVEDV and sPAP Echo/LVCO were moderately correlated with PVR( rs=0.62, 0.52, 0.63, both P<0.001). The ratio of sPAP Echo to LVEDV, when greater than or equal to 1.41, had a sensitivity of 0.800 and a specificity of 0.930 for determining PVR >1 000 dyn·s -1·cm -5 (AUC=0.860, P<0.001). Similarly, the ratio of sPAP Echo to LVIDd, when greater than or equal to 2.14, had a sensitivity of 0.647 and a specificity of 0.861 for determining PVR >1000 dyn·s -1·cm -5 (AUC=0.830, P<0.001). The sPAP Echo/LVIDd and mPAP Echo/LVIDd significantly decreased after PEA (both P<0.001). The sPAP Echo/LVIDd and mPAP Echo/LVIDd reduction rate (ΔsPAP Echo/LVIDd and ΔmPAP Echo/LVIDd) were significantly correlated with PVR reduction rate (ΔPVR), respectively ( rs=0.61, 0.63, both P<0.05). Conclusions:Two-dimensional ratio sPAP Echo/LVIDd and three-dimensional ratio sPAP Echo/LVEDV can be used to noninvasively estimate PVR in CTEPH patients. The conventional ratio sPAP Echo/LVIDd is convenient and reproducibly suitable for monitoring the improvement of PVR before and after treatment, and its ratio of 2.14 can predict the significant increase of PVR in CTEPH patients (>1 000 dyn·s -1·cm -5).
6.Customized Design and Three-Dimensional Molding of Bracketless Orthodontic Appliance
Tianmin GUAN ; Xin LI ; Qiang MA ; Yun ZHAI
Journal of Medical Biomechanics 2024;39(1):164-171
Objective A finite element simulation analysis of a bracketless orthodontic appliance was carried out to determine the correction amount under different working conditions.A design scheme for the bracketless orthodontic appliance was also formulated to create an orthodontic appliance formed by the fused deposition process with a personalized orthodontic effect.Methods Combined with the computed tomography data of the patient,a tooth model was reversely reconstructed.The correction amount of the tooth under tilting,twisting,and translation working conditions was established using the finite element method.Materials with different elastic moduli were chosen to make three-dimensional(3D)shaped bracketless orthodonitc appliance,and the orthodontic force was measured.Results Different correction amounts could be designed according to different working conditions so that the orthodontic appliance had a personalized treatment effect.The orthodontic force of the bracketless orthodontic appliance increased with elastic modulus and thickness.Under the given working conditions,the orthodontic force was minimal(90 mN)when the orthodontic material was at the minimum(90 mN).The orthodontic force reached its maximum value when the orthodontic material was at the maximum(61.66 N).Conclusions According to the patient's tooth condition and the size of the orthodontic force required for each step of correction in the whole correction process,the design should consider bracketless orthodontics with different elastic moduli and thicknesses to treat deformed teeth to realize the effect of staged orthodontic correction.
7.Effects of Zhuangyao Shuanglu Tongnao Formula on neuronal apoptosis of rats with ischemia-reperfusion induced injury
Yang ZHAI ; Xue-Ni MO ; Hong-Li TENG ; Yue-Qiang HU ; Guang-Shan ZHENG ; Wei MA ; Peng YANG ; Xiao-Ping MEI ; Min ZOU ; Kai-Hua WANG
Chinese Traditional Patent Medicine 2024;46(3):795-802
AIM To investigate the effects of Zhuangyao Shuanglu Tongnao Formula on neuronal apoptosis in rats with cerebral ischemia-reperfusion injury based on the study of oxidative stress and inflammatory response.METHODS The rats were randomly divided into the sham operation group,the model group,the edaravone group(3.0 mg/kg),the low,medium and high dose groups(9.0,18.0,36.0 g/kg)of Zhuangyao Shuanglu Tongnao Formula,with 18 rats in each group.The middle cerebral artery occlusion/reperfusion was conducted by thread embolism method to simulate cerebral ischemia reperfusion injury in rats followed by 6 days corresponding drugs administration.Subsequently,the rats had their neurological function deficit scored by Zeal Longa scoring method;their sizes of cerebral infarction areas measured by TTC staining;their pathological damage and apoptosis of neurons in hippocampal CA1 area of ischemic penumbra of the brain tissue detected by HE staining and TUNEL staining;their SOD activity and levels of GSH,MDA,IL-6,IL-1β,TNF-α in brain tissue detected by kits;and their protein expressions of Bax,Bcl-2,caspase-3,cleaved-capase-3,TLR4,NF-κB p65,Nrf2,HO-1 in rat brain tissue determined by Western blot.RESULTS Compared with the model group,the groups intervened with edaravone,medium and high dose of Zhuangyao Shuanglu Tongnao Formula displayed improvements in the scores of nerve function defects,the rate of cerebral infarction,the rate of neuronal apoptosis,the levels of IL-6,IL-1β,TNF-α and MDA in the ischemic penumbra of brain tissues,the protein expressions of Bax and TLR4,the ratio of cleaved-capase-3/caspase-3 and p-NF-κB p65/NF-κB p65(P<0.05),the levels of GSH,the activity of SOD and the protein expressions of Bcl-2,Nrf2 and HO-1(P<0.05).CONCLUSION Being an inhibitor of oxidative stress and inflammatory response,Zhuangyao Shuanglu Tongnao Formula can alleviate brain injury in rats with cerebral ischemia reperfusion injury through the inhibition of neuronal apoptosis and improvement of neural function mediated by the inhibition of TLR4/NF-κB signal pathway and activation of Nrf2/HO-1 signal pathway.
8.The role of preoperative intravenous administration of tranexamic acid in the treatment of proximal humeral fractures in elderly patients with locked steel plate internal fixation
Lei SHEN ; Panjun ZHANG ; Zhenhuan JIANG ; Chenjun ZHAI ; Tao JIANG ; Qiang WANG
Journal of Chinese Physician 2024;26(6):837-842
Objective:To explore and analyze the efficacy of preoperative intravenous tranexamic acid (TXA) in reducing bleeding and alleviating early postoperative pain in elderly patients with proximal humeral fractures (PHF) who underwent minimally invasive plate osteosynthesis (MIPO) approach with open reduction and locking plate internal fixation.Methods:A retrospective analysis was conducted on the data of 165 elderly patients with partial or partial PHF who underwent open reduction and locking plate internal fixation via MIPO approach at the Yixing People′s Hospital from June 2018 to June 2021. According to whether TXA was used intravenously 30 minutes before surgery, patients were divided into a TXA group (77 cases) and a control group (88 cases). The surgical time, hemoglobin decrease, total blood loss (TBL), intraoperative blood loss (IBL), postoperative drainage volume, visible blood loss (VBL), hidden blood loss (HBL), blood transfusion status, Visual Analogue Scale (VAS) score for surgical site pain 24 hours after surgery, postoperative hospital stay, shoulder Constant-Murley function score at 1 and 3 months after surgery, and complications were recorded and compared between the two groups.Results:The TBL, VBL, IBL, HBL, postoperative drainage volume, hemoglobin decrease, transfusion rate, postoperative VAS score at the surgical site, and hospital stay in the TXA group were all lower than those in the control group, and the differences were statistically significant (all P<0.05). The VAS score at the surgical site 24 hours after surgery was positively correlated with TBL and HBL in two groups of patients ( r=0.402, 0.418, P<0.001). Compared with the control group, the TXA group had a higher shoulder Constant-Murley function score at 1 month after surgery, and the difference was statistically significant ( P=0.002). There was no statistically significant difference in shoulder Constant-Murley function score and incidence of complications between the two groups at 3 months after surgery (all P>0.05). Conclusions:For elderly patients with partial or partial PHF who underwent open reduction and locking plate internal fixation using MIPO approach, intravenous infusion of TXA 30 minutes before surgery can help alleviate postoperative pain at the surgical site, reduce intraoperative and postoperative bleeding, lower transfusion rates, shorten hospital stay, and do not increase the incidence of complications such as incision abnormalities and thrombosis, which is beneficial for promoting early and rapid recovery of patients.
9.Analysis of risk factors for the occurrence and aggravation of lower back pain in Parkinson′s disease patients
Lei SHEN ; Qiang WANG ; Yitong XIONG ; Junfeng SHI ; Zhenhuan JIANG ; Chenjun ZHAI ; Tao JIANG
Journal of Chinese Physician 2024;26(9):1322-1327
Objective:To explore the independent risk factors for the occurrence and aggravation of lower back pain (LBP) in patients with Parkinson′s disease (PD), in order to provide reference for clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the case data of 309 PD patients who visited the Affiliated Yixing Hospital of Jiangsu University from June 2018 to May 2020. The KING Parkinson′s Disease Pain Scale (KPPS) was used to quantitatively evaluate the LBP of PD patients, who were divided into LBP group and Non LBP group. The general clinical data, PD related data, and imaging data of the two groups were compared and analyzed. Binary logistic regression analysis was used to evaluate independent risk factors for LBP in PD patients. Pearson correlation analysis was conducted between KPPS scores and various factors, and linear regression analysis was used to identify the relevant risk factors that exacerbate LBP in PD patients.Results:Compared with the Non LBP group, the LBP group had lower bone mineral density (BMD) and a lower proportion of patients who engaged in daily exercise. The difference between the two groups was statistically significant (all P<0.05). Compared with the Non LBP group, patients in the LBP group had a longer course of illness, higher stiffness scores, a higher proportion of patients with fluctuating symptoms, higher UPDRS-Ⅲ scores, and a higher proportion of patients with thoracolumbar fascial injury (TLFI) and lumbar sagittal imbalance. The differences between the two groups were statistically significant (all P<0.05). The results of binary logistic regression analysis showed that combined TLFI ( OR=2.773, 95% CI: 1.219-6.309, P=0.015), combined lumbar sagittal imbalance ( OR=4.835, 95% CI: 2.244-10.421, P<0.001), and lower BMD ( OR=2.818, 95% CI: 1.767-4.493, P<0.001) were risk factors for LBP in PD patients. The KPPS score was correlated with BMD and TLFI ( r=-0.146, 0.294, all P<0.05). The linear regression results showed that the merged TLFI ( B=2.271, β=0.285, P<0.001) was positively correlated with KPPS score, indicating a risk factor. Conclusions:The combination of TLFI, lumbar sagittal imbalance, and lower BMD is closely related to the occurrence of LBP in PD patients, and the combination of TLFI is an independent risk factor for exacerbating LBP symptoms. Clinical attention should be paid to the prevention and treatment of TLFI in PD patients.
10.Regulatory effect and mechanism of total alkaloids of Rhizoma Corydalis on hippocampal silencing information regulator 1/tumor suppressor P53 protein signaling pathway in rats with chronic cerebral ischemia
Jun LI ; Yazhi QI ; Ya TANG ; Rui CAO ; Yanling ZHAI ; Yusheng HAN ; Qiang XU
Chinese Journal of Cerebrovascular Diseases 2023;20(12):837-845
Objective To investigate the effect of total alkaloids of Rhizoma Corydalis(TAC)on the expression of silencing information regulator 1(Sirt1)/tumor suppressor P53 protein signaling pathway-related proteins in the hippocampus of rats with chronic cerebral ischemia(CCH),and to explore its mechanism.Methods The rats were randomly divided into Sham operation group,model group,TAC high-dose group(14 mg/kg)and TAC low-dose group(7 mg/kg),with 6 rats in each group.A modified bilateral common carotid artery permanent occlusion method(BCCAO)was used to establish a rat model of CCH,and only bilateral common carotid arteries were separated in the Sham group.After the modeling was completed,each group was given the corresponding drug or isotonic saline by gavage,once a day,and the treatment lasted for 14 days.Hematoxycin-eosin staining was used to observe the pathological changes in the hippocampus of rats,in situ terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphate-biotin nick end labeling assay(TUNEL)was used to detect neuronal apoptosis,and Western blotting and immunohistochemistry were used to detect expression of Sirt1,P53,P53 positive apoptosis regulator(PUMA),B-cell lymphocytoma-2(Bcl-2)protein,Bcl-2-related X protein(BAX),respectively in the hippocampus of rats.Results(1)There were significant differences in the number of apoptotic cells and apoptosis rate among the four groups(F-values were 71.417 and 76.835,respectively,both P<0.01).There were statistically significant differences in the mean integral optical density values of Sirt1,P53,PUMA,BAX and Bcl-2 protein positive expression areas among the four groups(F-values were 1 178.390,42.465,867.413,110.656 and 131.801,all P<0.01).There were significant differences in the relative expression levels of Sirt1,P53,PUMA,BAX and Bcl-2 among the four groups(F-values were 9.497,11.863,58.552,186.855 and 12.466,all P<0.01).(2)Compared with the Sham operation group,the neuronal arrangement of brain tissue in the hippocampus of the model group was disordered,the nuclear consolidation increased,and the glial cells and inflammatory cells increased significantly,and the number and apoptosis rate of neurons in the hippocampus of the model group increased significantly(respectively[10.8±1.5]cells vs.[2.0±0.9]cells and[35.5±4.5]%vs.[6.2±2.6]%;both P<0.05),and the average integral optical density values of the positive expression areas of Sirt1 and Bcl-2 proteins decreased significantly(84.6±6.6 vs.244.6±4.9,138.5±6.7 vs.210.9±10.0;both P<0.05),the average integral optical density values of P53,PUMA and BAX proteins were significantly increased(156.8±11.6 vs.93.5±11.6,151.3±3.3 vs.38.0±4.0,87.0±5.0 vs.38.4±5.5;all P<0.05),the relative expression levels of Sirt1 and Bcl-2 proteins were significantly decreased(0.51±0.07 vs.0.74±0.07,0.36±0.03 vs.0.53±0.05;both P<0.05),and the relative expression levels of P53,PUMA and BAX proteins were significantly increased(0.37±0.06 vs.0.21±0.02,0.62±0.06 vs.0.23±0.02,1.08±0.06 vs.0.45±0.03;all P<0.05).(3)Compared with the model group,the hippocampal tissue structure of the high-dose and low-dose TAC groups was relatively compact and uniform,the neurons were neatly arranged,and the cell structure was relatively clear and complete,while the number of neuronal apoptotic cells and the apoptosis rate decreased significantly(respectively[3.8±0.7]cells vs.[6.2±1.2]cells,[12.4±2.8]%vs.[20.2±3.9]%;both P<0.05),and the average integrated optical density values of the positive expression areas of Sirt1 and Bcl-2 proteins(the high-dose and low-dose TAC groups:Sirt1 150.0±4.8,131.3±1.3,and Bcl-2 207.1±7.4,169.5±3.9,respectively)were significantly increased(both P<0.05),the average integral optical density values of P53,PUMA and BAX proteins were significantly decreased(the high-dose and low-dose TAC groups:P53 105.9±8.8,115.5±9.0,and PUMA56.8±5.1,74.4±3.9,and BAX40.5±5.6,48.4±5.0,respectively,all P<0.05),the relative expression levels of Bcl-2 protein(the high-dose and low-dose TAC groups:0.53±0.05,0.47±0.02,respectively)were significantly increased(P<0.05),the relative expression levels of P53(the high-dose and low-dose TAC groups:0.21±0.02,0.24±0.04,respectively),PUMA(the high-dose and low-dose TAC groups:0.36±0.02,0.28±0.04,respectively)and BAX proteins(the high-dose and low-dose TAC groups:0.52±0.02,0.54±0.03,respectively)were significantly decreased(all P<0.05),the relative expression level of Sirt1 protein in the TAC high-dose group was significantly decreased(0.71±0.05,P<0.05),and the relative expression level of Sirt1 protein in the TAC low-dose group was not statistically significant(0.52±0.08,P>0.05).Conclusion TAC can alleviate neuronal damage and reduce the apoptosis rate of neurons in the hippocampus of CCH rats,and the mechanism may be related to the activation of Sirt1/P53 pathway,inhibition of P53 protein activity,and thus the expression level of apoptosis-related proteins in the downstream of TAC.

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