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1.
Acta Pharmaceutica Sinica ; (12): 2853-2861, 2023.
Article in Chinese | WPRIM | ID: wpr-999035

ABSTRACT

There is still a serious challenge of the measurement of critical quality attributes (CQAs) related to clinical efficacy for Chinese materia medica manufacturing. To overcome this challenge, an integrated strategy of biosensor and ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) was proposed using Tongren niuhuang qingxin pills as a trial. Firstly, an original biosensor was created using a semiconductor chip material high electron mobility transistor (HEMT) as the transducer and the macrophage migration inhibitory factor (MIF) as the identification element. By this MIF-HEMT biosensor, the efficacy on stoke of different components from Tongren niuhuang qingxin pills was measured. It was clear that all three components of Tongren niuhuang qingxin pills had strong therapeutic effects on stroke, especially the section A, the KD of which reached to 8.722×10-10 g·mL-1. Furthermore, MIF-HEMT biosensor integrated UPLC-MS/MS was introduced to identify the efficacy CQAs of different components of Tongren niuhuang qingxin pills. As a result, 19 potential CQAs, such as albiforin, paeoniflorin, and prim-O-glucosylcimifugin, were measured as the efficacy CQAs of Tongren niuhuang qingxin pills on stroke treatment by MIF. These results provided vital measurement techniques and methodological guidance for the CQAs study of Tongren niuhuang qingxin pills intervention in MIF-induced stroke treatment. This also provided an essential guideline for the efficient utilization and quality control measurement of high-quality classical recipes.

2.
Chinese Pharmacological Bulletin ; (12): 2205-2210, 2023.
Article in Chinese | WPRIM | ID: wpr-1013663

ABSTRACT

Inflammatory bowel disease (IBD), as an idiopathic inflammatory disease of the intestinal tract, consisting mainly of Crohns disease and ulcerative colitis, which can involve the rectum, colon and ileum, and whose pathogenesis is still not fully understood. The initiation of intestinal inflammation associated with IBD and its chronieity begins with increased intestinal permeability caused by intestinal epithelial barrier disruption. The anti-permeability of the intestinal epithelial barrier is maintained by tight junction in the apical region of the intestinal epithelial cells, and disruption of the tight junction structure is closely associated with intestinal epithelial barrier damage and the development of IBD. Therefore, it is significant to find drugs for the prevention and treatment of IBD using tight junctions as regulatory targets. In recent years, many small molecules of natural product origin have been reported to improve the effects of IBD. In particular, we review the compounds that have the function of repairing intestinal epithelial barrier and protecting tight junction structure, in order to provide research ideas for the design and development of new drugs for the prevention and treatment of IBD.

3.
China Journal of Chinese Materia Medica ; (24): 4702-4710, 2023.
Article in Chinese | WPRIM | ID: wpr-1008637

ABSTRACT

This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine.


Subject(s)
Humans , Carcinoma, Hepatocellular/genetics , Proto-Oncogene Proteins c-akt/metabolism , Caspase 3/metabolism , Liver Neoplasms/genetics , Molecular Docking Simulation , Sincalide/pharmacology , Cell Line, Tumor , Cell Proliferation , Hep G2 Cells , TOR Serine-Threonine Kinases/metabolism , Apoptosis
4.
China Journal of Chinese Materia Medica ; (24): 5337-5344, 2023.
Article in Chinese | WPRIM | ID: wpr-1008731

ABSTRACT

This study aims to explore the effects of Shenqi Dihuang Decoction on high-glucose induced ferroptosis and the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4) axis in human renal tubular epithelial cells(HK-2) and to clarify the underlying mechanism. The cell injury model was established by exposing HK-2 to high glucose, and the Shenqi Dihuang Decoction-medicated serum was prepared. The optimal concentration and intervention time of Shenqi Dihuang Decoction were determined. HK-2 were divided into normal, high glucose, and low-, medium-, and high-dose Shenqi Dihuang Decoction groups. After interventions, the cell proliferation rate in each group was determined and the cell morphology and mitochondrial ultrastructure were observed. Then, the levels of intracellular reactive oxygen species(ROS), ferrous ion(Fe~(2+)), glutathione(GSH), and malondialdehyde(MDA) and the protein levels of Nrf2, HO-1, GPX4, and xCT were measured. The optimal concentration and intervention time of Shenqi Dihuang Decoction-medicated serum were determined to be 10% and 24 h, respectively. Compared with the high glucose group, high-dose Shenqi Dihuang Decoction promoted the proliferation of HK-2. The cells in the low-, medium-, and high-dose Shenqi Dihuang Decoction groups presented tight arrangement, an increased cell count, improved morphology from a spindle-fiber shape to a cobblestone shape, and improved morphology and structure of mitochondrial membrane and cristae, compared with those in the high glucose group. Meanwhile, all the doses of Shenqi Dihuang Decoction inhibited ROS elevation to mitigate the peroxidation damage, lowered the Fe~(2+) and MDA levels and elevated the GSH level to inhibit lipid peroxidation, and activated the antioxidant pathway to upregulate the protein levels of Nrf2, HO-1, xCT, and GPX4. In conclusion, Shenqi Dihuang Decoction-medicated serum can inhibit high-glucose induced ferroptosis of HK-2 in vitro, which involves the antioxidant effect and the activation of the Nrf2/HO-1/GPX4 pathway.


Subject(s)
Humans , Ferroptosis , NF-E2-Related Factor 2/genetics , Reactive Oxygen Species , Epithelial Cells , Antioxidants , Glutathione , Glucose
5.
China Journal of Chinese Materia Medica ; (24): 4475-4482, 2023.
Article in Chinese | WPRIM | ID: wpr-1008702

ABSTRACT

This study investigated the effect and mechanism of morin in inducing autophagy and apoptosis in hepatocellular carcinoma cells through the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription protein 3(STAT3) pathway. Human hepatocellular carcinoma SK-HEP-1 cells were stimulated with different concentrations of morin(0, 50, 100, 125, 200, and 250 μmol·L~(-1)). The effect of morin on the viability of SK-HEP-1 cells was detected by Cell Counting Kit-8(CCK-8). The effect of morin on the proliferation and apoptosis of SK-HEP-1 cells was investigated using colony formation assay, flow cytometry, and BeyoClick~(TM) EdU-488 with different concentrations of morin(0, 125, and 250 μmol·L~(-1)). The changes in the autophagy level of cells treated with morin were examined by transmission electron microscopy and autophagy inhibitors. The impact of morin on the expression levels of proteins related to the Akt/mTOR/STAT3 pathway was verified by Western blot. Compared with the control group, the morin groups showed decreased viability of SK-HEP-1 cells in a time-and concentration-dependent manner, increased number of apoptotic cells, up-regulated expression level of apoptosis marker PARP, up-regulated phosphorylation level of apoptosis-regulating protein H2AX, decreased number of positive cells and the colony formation rate, an upward trend of expression levels of autophagy-related proteins LC3-Ⅱ, Atg5, and Atg7, and decreased phosphorylation levels of Akt, mTOR, and STAT3. These results suggest that morin can promote apoptosis, inhibit proliferation, and induce autophagy in hepatocellular carcinoma cells, and its mechanism of action may be related to the Akt/mTOR/STAT3 pathway.


Subject(s)
Humans , Proto-Oncogene Proteins c-akt/metabolism , Carcinoma, Hepatocellular/pathology , Liver Neoplasms/pathology , TOR Serine-Threonine Kinases/metabolism , Apoptosis , Autophagy , Cell Proliferation , Cell Line, Tumor , STAT3 Transcription Factor/metabolism
6.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 572-577, 2022.
Article in Chinese | WPRIM | ID: wpr-936258

ABSTRACT

Objective: To investigate the feasibility, safety and efficacy of transoral robotic surgery (TORS) in the treatment of lingual thyroglossal duct cyst (LTGDC). Methods: The clinical data of 10 patients with LTGDC treated with TORS in Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from May 2017 to November 2020 were analyzed retrospectively,including 6 males and 4 females, aged 5-44 years. The cysts were fully exposed, and resection usually started from the cephalic side of lesions. The range of resection was 3 to 5 mm away from the lesions, and partial hyoid bone was removed if necessary. Intra-operative robotic set-up time,operation time and estimated blood loss,and post-operative local bleeding, dyspnea and recovery time for oral intake were analyzed. SPSS 12.0 software was used for statistical analysis. Results: The cysts in all 10 patients were successfully resected by TORS with da Vinci Si surgical system. The mean robotic set-up and exposure time, operation time, estimated intraoperative blood loss and recovery time for oral intake were (15.5±7.1) min, (17.6±7.4) min, (8.9±6.4)ml and (2.3±2.2)days, respectively. No patient required tracheostomy intra-or post-operatively, and no symptoms of airway obstruction, postoperative bleeding, pharyngeal fistula, hoarseness and neurological impairment occurred after operation. The patients were followed up for 5 to 47 months, with median follow-up time of 17 months, and no recurrence was observed. Conclusion: TORS is safe and feasible for resection of LTGDC, with rapid recovery and low recurrence rate.


Subject(s)
Female , Humans , Male , Retrospective Studies , Robotic Surgical Procedures , Robotics , Thyroglossal Cyst/surgery , Tongue/surgery , Treatment Outcome
7.
China Journal of Chinese Materia Medica ; (24): 3422-3428, 2021.
Article in Chinese | WPRIM | ID: wpr-887993

ABSTRACT

The effect of Danhong Injection on the endogenous metabolites of rabbit platelets was analyzed by the liquid chromatography-mass spectrometry( LC-MS) based metabonomic approach. Anti-platelet aggregation was detected after Danhong Injection treatment and the changes of platelet metabolites were analyzed by metabonomics. Principal component analysis( PCA) and partial least squares discriminant analysis( PLS-DA) were performed to investigate the effect of Danhong Injection on endogenous metabolites of platelets,characterize the biomarkers,and explore the relevant pathways and the underlying mechanism. As demonstrated by the pharmacodynamic results,Danhong Injection of different doses and concentrations antagonized platelet aggregation in a dose-and concentration-dependent manner. In contrast to the control group,25 differential metabolites such as nicotinic acid,nicotinic acid riboside,and hypoxanthine were screened out after platelets were treated by Danhong Injection. These metabolites,serving as important biomarkers,were mainly enriched in the nicotinic acid-niacinamide metabolic pathway and purine metabolic pathway. This study explored the therapeutic mechanism of Danhong Injection from a microscopic perspective by metabonomics,which is expected to provide a new idea for the investigation of platelet-related mechanisms.


Subject(s)
Animals , Rabbits , Biomarkers , Blood Platelets , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/pharmacology , Metabolomics , Technology
8.
China Journal of Chinese Materia Medica ; (24): 2276-2286, 2021.
Article in Chinese | WPRIM | ID: wpr-879188

ABSTRACT

The metabolites of salvianolic acid A and salvianolic acid B in rats were analyzed and compared by ultra-high-perfor-mance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS). After the rats were administrated by gavage, plasma at different time points and urine within 24 hours were collected to be treated by solid phase extraction(SPE), then they were gradient eluted by Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) and 0.1% formic acid solution(A)-acetonitrile(B) mobile phase system, and finally all biological samples of rats were analyzed under negative ion scanning mode. By obtaining the accurate relative molecular mass and multi-level mass spectrometry information of metabolites, combined with the characteristic cleavage law of the reference standard and literature reports, a total of 30 metabolites, including salvianolic acid A and B, were identified. Among them, there were 24 metabolites derived from salvianolic acid A, with the main metabolic pathways including ester bond cleavage, dehydroxylation, decarboxylation, hydrogenation, methylation, hydroxylation, sulfonation, glucuronidation, and their multiple reactions. There were 15 metabolites of salvianolic acid B, and the main biotransformation pathways were five-membered ring cracking, ester bond cleavage, decarboxylation, dehydroxylation, hydrogenation, methylation, sulfonation, glucuronidation, and their compound reactions. In this study, the cross-metabolic profile of salvianolic acid A and B was elucidated completely, which would provide reference for further studies on the basis of pharmacodynamic substances and the exploration of pharmacological mechanism.


Subject(s)
Animals , Rats , Benzofurans , Caffeic Acids , Chromatography, High Pressure Liquid , Lactates , Mass Spectrometry , Technology
9.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 109-115, 2020.
Article in Chinese | WPRIM | ID: wpr-787616

ABSTRACT

To evaluate the indication, safety and effectiveness of transoral robotic surgery (TORS) for oropharyngeal cancer based on our preliminary experience. Twelve patients, including six with tonsil cancer, five with tongue base cancer and one with posterior pharyngeal wall cancer, who underwent TORS with Da Vinci Si surgical system from March 2017 to October 2018 at Tongji Hospital of Huazhong University of Science Technology were respectively analyzed. And the surgical time, intraoperative blood loss, postoperative local bleeding, dyspnea, nerve function injury, oral intake time, whether or not to receive chemoradiotherapy were analyzed. All tumors in the 12 patients were en bloc removed by TORS. Surgical time ranged from 25 to 80 min with an average of 34.2 min. The blood loss ranged from 10 ml to 50 ml with an average of 20.8 ml. The recovery time for oral intake ranged from 1 day to 30 days with an average of 8.4 days. No patient underwent tracheostomy after TORS. Also, no patient manifested with airway obstruction, bleeding or nerve injury symptoms after operation. All 12 patients reached pathologically negative surgical margins. The patients were followed up for 4 to 22 months, with a median of 12 months. All patients who combined with more advanced than T3 stage, or more advanced than N2 stage were recommended to oncologist, then, followed with radiotherapy or chemoradiotherapy if no relevant contradictions occurred. No local recurrence or distant metastasis case was found. With proper indications, the application of TORS in oropharyngeal cancer is a relatively safe, effective and minimal invasive therapy, which merits more clinical applications.

10.
Acta Anatomica Sinica ; (6): 502-506, 2020.
Article in Chinese | WPRIM | ID: wpr-1015537

ABSTRACT

Objective Exploring the effect of spinal cord decellularized scaffold on spinal cord defects and observing the behavior and regeneration of rats after operation. Methods The spinal cords of 30 SD rats were treated with 3% Triton X-100 and 2% sodium deoxycholate on oscillator. The cell residue and the spatial structure of the tissue were compared before and after treatment, in order to understand the tissue structure of the stent itself. 90 SD rats were randomly divided into control group, simple injury group and stent transplantation group. Excision of the spinal cord 9-10 segments in the simple injury group and the stent graft group the acellular scaffold was transplanted to the stent graft group. Behavioral scores were observed postoperatively. At 4, 8, and 12 weeks, the spinal cords of the injured part of the rats were taken for HE staining and immunofluorescence detection of nerve regeneration-related proteins. Results After decellularization of the spinal cord, the nerve cells and axons were completely removed, and the extracellular matrix of the spinal cord was preserved. Scanning electron microscopy revealed that the scaffold retained a certain porous network scaffold structure. In the experiment of decellularized scaffold in vivo, the Basso-Beattie-Bresnahan(BBB) score showed that the recovery of hindlimb motor function in rats with decellularized scaffolds was better than that in rats with simple injury. HE staining showed that the decellularized scaffold could fill the defect of the spinal cord segment and accelerate the repair process of the injured spinal cord. Immunofluorescence showed that there was a certain axonal regeneration in the injured part of the stent transplantation group. Conclusion The spinal cord decellularized scaffold retains the extracellular matrix and has a certain spatial structure, which can accelerate the process of spinal cord defect repair to a certain extent, and has a certain promoting effect on nerve regeneration.

11.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 65-71, 2020.
Article in Chinese | WPRIM | ID: wpr-873219

ABSTRACT

Objective:To explore the effect and mechanism of esculentoside on lipopolysaccharide-induced mastitis in mice. Method:Female BALB/c mice were randomly divided into control group, model group, dexamethasone group (DEX, 5.0 mg·kg-1) and esculentoside group (50, 25, 12.5 mg·kg-1). The mastitis model of postpartum female mice (BALB/c) induced by lipopolysaccharide (LPS) was used to analyze the pathological conditions of breast tissue and the activity of recombinant myeloperoxidase(MPO), factor content and oxidative stress level. Western blot was used to evaluate the effect of esculentoside on Toll-like receptor4 (TLR4)/nuclear transcription factor (NF-κB) signaling pathway proteins. Result:Compared with the normal group, the breast tissue of the model group had typical mastitis changes, such as hyperemia and congestion, the level of MPO increased, and the expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) increased significantly (P<0.01). Compared with LPS model group, esculentoside groups could significantly improve the inflammatory damage of mammary gland tissue, reduce the secretion of neutrophils and the activity of MPO, the expression levels of pro-inflammatory factors TNF-α, IL-1β and IL-6 were also significantly down-regulated, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were increased, while the level of malondialdehyde (MDA) was decreased, and the activation of TLR4/NF-κB signaling pathway was inhibited by esculentoside(P<0.05,P<0.01). Conclusion:Esculentoside have a protective effect on lipopolysaccharide-induced mastitis in mice, which may be related to the inhibition of TLR4/NF-κB signaling pathway protein expression.

12.
Journal of Experimental Hematology ; (6): 1957-1961, 2020.
Article in Chinese | WPRIM | ID: wpr-879999

ABSTRACT

OBJECTIVE@#To investigate the expression of immunoglobulin G (IgG) and its subclasses in patients with multiple myeloma (MM) and lymphoma (LYM) and its correlation with blood cell parameters.@*METHODS@#129 patients with multiple myeloma and 113 patients with lymphoma diagnosed treated in Sichuan people's Hospital from January to December 2019 were selected and the total IgG and subclass IgG1, IgG2, IgG3 and IgG4, and some parameters of peripheral blood routine in patients were retrospective analyzed. Independent sample t test or nonparametric Mann Whitney U test were used for comparison between the groups. The relationship between IgG subclass and blood cell parameters was analyzed by correlation analysis. For the bivariate normal distribution data, Pearson correlation coefficient was calculated. For bivariate non normal distribution data, the Spearman correlation coefficient was calculated.@*RESULTS@#IgG1 and IgG2 were significantly higher in patients with multiple myeloma than in patients with lymphoma (P=0.001, 0.000, respectively), but IgG3 and IgG4 were significantly lower than in patients with lymphoma (P=0.000, 0.000, respectively), and there was no significant difference for total serum IgG between the two groups (P=0.717). The results showed that the IgG4 content of male patients with multiple myeloma and lymphoma was significantly higher than that of female patients (Z=-3.191, P=0.001); the age, M%, E% of the MM patients were significantly higher than those of the patients with lymphoma (P=0.000, 0.005, 0.019), but WBC, RBC, Hb were significantly lower than those of the patients with lymphoma (P=0.013, 0.000, respectively). The results of Spearman correlation analysis showed that RBC and Hb were decreased with the increasing of IgG and IgG1 in the MM patients(r=0.254, -0.272, -0.248 and -0.289, P=0.004, 0.002, 0.005 and 0.001). With the increasing of IgG4 in the serum of patients with lymphoma, RBC and Hb showed the trendy of decreased (r=-0.240 and, -0.251, respectively, P=0.010, 0.007).@*CONCLUSION@#The detection of IgG subclass and the correlation between IgG subclass and blood cell parameters are great value in the diagnosis and pathogenesis of multiple myeloma and lymphoma.


Subject(s)
Female , Humans , Male , Blood Cells , Immunoglobulin G , Lymphoma , Multiple Myeloma , Patients , Retrospective Studies , Serum
13.
China Journal of Chinese Materia Medica ; (24): 3952-3960, 2020.
Article in Chinese | WPRIM | ID: wpr-828362

ABSTRACT

A method of ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field Obitrap high-resolution mass spectrometry(UHPLC-Q-Exactive MS) was established to comprehensively identify the metabolites of carnosic acid in rats. After oral gavage of carnosic acid CMC-Na suspension in rats, urine, plasma and feces samples were collected and pretreated by solid phase extraction(SPE). Acquity UPLC BEH C_(18 )column(2.1 mm×100 mm, 1.7 μm) was used with 0.1% formic acid solution(A)-acetonitrile(B) as the mobile phase for the gradient elution. Biological samples were analyzed by quadrupole/electrostatic field Obitrap high-resolution mass spectrometry in positive and negative ion mode. Based on the accurate molecular mass, fragment ion information, and related literature reports, a total of 28 compounds(including carnosic acid) were finally identified in rat samples. As a result, the main metabolic pathways of carnosic acid in rats are oxidation, hydroxylation, methylation, glucuronide conjugation, sulfate conjugation, S-cysteine conjugation, glutathione conjugation, demethylation, decarbonylation and their composite reactions. The study showed that the metabolism of carnosic acid in rats could be efficiently and comprehensively clarified by using UHPLC-Q-Exactive MS, providing a reference for clarifying the material basis and metabolic mechanism of carnosic acid.


Subject(s)
Animals , Rats , Abietanes , Chromatography, High Pressure Liquid , Mass Spectrometry , Solid Phase Extraction
14.
Chinese Medical Journal ; (24): 1192-1202, 2020.
Article in English | WPRIM | ID: wpr-827628

ABSTRACT

BACKGROUND@#Pulmonary fibrosis is a respiratory disease caused by the proliferation of fibroblasts and accumulation of the extracellular matrix (ECM). It is known that the lung ECM is mainly composed of a three-dimensional fiber mesh filled with various high-molecular-weight proteins. However, the small-molecular-weight proteins in the lung ECM and their differences between normal and fibrotic lung ECM are largely unknown.@*METHODS@#Healthy adult male Sprague-Dawley rats (Rattus norvegicus) weighing about 150 to 200 g were randomly divided into three groups using random number table: A, B, and C and each group contained five rats. The rats in Group A were administered a single intragastric (i.g.) dose of 500 μL of saline as control, and those in Groups B and C were administered a single i.g. dose of paraquat (PQ) dissolved in 500 μL of saline (20 mg/kg). After 2 weeks, the lungs of rats in Group B were harvested for histological observation, preparation of de-cellularized lung scaffolds, and proteomic analysis for small-molecular-weight proteins, and similar procedures were performed on Group C and A after 4 weeks. The differentially expressed small-molecular-weight proteins (DESMPs) between different groups and the subcellular locations were analyzed.@*RESULTS@#Of the 1626 small-molecular-weight proteins identified, 1047 were quantifiable. There were 97 up-regulated and 45 down-regulated proteins in B vs. A, 274 up-regulated and 31 down-regulated proteins in C vs. A, and 237 up-regulated and 28 down-regulated proteins identified in C vs. B. Both the up-regulated and down-regulated proteins in the three comparisons were mainly distributed in single-organism processes and cellular processes within biological process, cell and organelle within cellular component, and binding within molecular function. Further, more up-regulated than down-regulated proteins were identified in most sub-cellular locations. The interactions of DESMPs identified in extracellular location in all comparisons showed that serum albumin (Alb) harbored the highest degree of node (25), followed by prolyl 4-hydroxylase beta polypeptide (12), integrin β1 (10), apolipoprotein A1 (9), and fibrinogen gamma chain (9).@*CONCLUSIONS@#Numerous PQ-induced DESMPs were identified in de-cellularized lungs of rats by high throughput proteomics analysis. The DESMPs between the control and treatment groups showed diversity in molecular functions, biological processes, and pathways. In addition, the interactions of extracellular DESMPs suggested that the extracellular proteins Alb, Itgb1, Apoa1, P4hb, and Fgg in ECM could be potentially used as biomarker candidates for pulmonary fibrosis. These results provided useful information and new insights regarding pulmonary fibrosis.

15.
China Journal of Chinese Materia Medica ; (24): 4713-4719, 2019.
Article in Chinese | WPRIM | ID: wpr-1008250

ABSTRACT

The project was launched to analyze the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats by metabonomics. The preparation method of sulfur-fumigated Ophiopogonis Radix in laboratory was established. Then the blood samples of SD rats in blank group,Ophiopogonis Radix extract group and sulfur-fumigated Ophiopogonis Radix extract group were investigated by UHPLC-Q-Exactive. The differential metabolites were screened and identified by PCA(principal component analysis),OPLSDA(orthogonal partial least squares discriminant analysis) and variable importance projection(VIP),and the metabolic pathways were analyzed. Finally,a total of 15 potential biomarkers were identified. Compared with the samples of Ophiopogonis Radix extract group,sulfur-fumigated Ophiopogonis Radix mainly affected the biosynthesis and metabolism of amino acids in normal rats. Its mechanism may be related to the biosynthesis of phenylalanine,tyrosine,tryptophan and aminoacyl-tRNA as well as the metabolism of phenylalanine and tryptophan. Based on UHPLC-HRMS metabonomics,this paper discussed the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats,which provided an idea for the metabolic study of other sulfur-fumigated traditional Chinese medicines.


Subject(s)
Animals , Rats , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Metabolomics , Rats, Sprague-Dawley , Sulfur
16.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 401-412, 2019.
Article in English | WPRIM | ID: wpr-776870

ABSTRACT

Atherosclerosis (AS) is a chronic inflammatory disease associated with high morbidity and mortality. The incidence of AS is increasing in the last decades. So development of safe and effective therapeutics for treating AS has become prominently important. Although there are numerous chemical drugs available for treating AS, some drugs are not effective and some have serious side effects. Traditional Chinese medicine (TCM) has a long history for the prevention and treatment of AS due to its less side effects and superior efficacy. This paper describes the effectiveness and underlying mechanisms for prevention and treatment of AS by TCM or its active components. Some TCM, e.g. XuemaiNing, Tongxinluo and Salvia miltiorrhiza have been reported to have cardio-protective effect. Some active components of TCM, e.g. saikosaponin-A, kuwanon G, luteolin and β-elemene have been isolated from various TCM and demonstrated to have beneficial effects on prevention and treatment of AS.


Subject(s)
Animals , Humans , Atherosclerosis , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Medicine, Chinese Traditional , Plants, Medicinal , Chemistry
17.
Acta Pharmaceutica Sinica ; (12): 2184-2188, 2019.
Article in Chinese | WPRIM | ID: wpr-780341

ABSTRACT

A bioassay method for inhibiting platelet aggregation in vitro was established to quantify the pharmacological effects of Compound Danshen Tablets and support its quality control. The inhibition of platelet aggregation in rabbit plasma in vitro by Compound Danshen Tablets was used as the experimental system. The titer was calculated by using the method of dose-response parallel lines. As a result, a bioassay for the inhibition of platelet aggregation in vitro by Compound Danshen Tablets was established. Linearity was good in the concentration range of 0.128 g·mL-1 to 0.205 g·mL-1, and the titer of standard Compound Danshen tablets was 7 659 U·g-1 according to the titer definition. This in vitro assay was simple, reliable, reproducible and convenient. The activity of Compound Danshen Tablets in inhibiting platelet aggregation was quantified by potency assay and the quality of different batches was evaluated. The method can be applied for the quality control of Compound Danshen Tablets.

18.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 181-186, 2019.
Article in Chinese | WPRIM | ID: wpr-802117

ABSTRACT

Preliminary results have been achieved in the prevention and treatment of cardiovascular disease (CVD) in China, but the number of CVD patients will continue to grow rapidly in the next 10 years. Generally speaking, the prevalence and mortality of CVD in China are still on the rise. CVD has worsened and become a major public health problem. It is urgent to prevent CVD. Therefore, the development of safe, effective and inexpensive drugs for the prevention and treatment of CVD is a livelihood project of great significance. Allii Sativi Bulbus, as a commonly used medicinal and edible herbal medicine, has rich resources and a long history of application. Since the 1980s, many studies on Allii Sativi Bulbus have been carried out by global scholars. The results show that Allii Sativi Bulbus has the effects in reducing blood lipid, preventing atherosclerosis, lowering blood pressure, resisting myocardial ischemia and treating other cardiovascular diseases, with many pharmacological activities such as anti-tumor, anti-bacterial, anti-inflammatory, insecticidal, regulating immunity, anti-oxidation, free radical scavenging activities. Focusing on the regulation of blood lipid metabolism by Allii Sativi Bulbus, the latest literatures at home and abroad have been consulted. Both basic research and clinical trials have shown that Allii Sativi Bulbus has a good hypolipidemic effect. Its mechanism is mainly related to the inhibition of key enzymes in the process of lipid synthesis and related factors regulating lipid metabolism. In this paper, the hypolipidemic effect and molecular mechanism of garlic were reviewed to define its research trends and problems, and provide reference for further research and clinical application of Allii Sativi Bulbus.

19.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 163-167, 2019.
Article in Chinese | WPRIM | ID: wpr-802047

ABSTRACT

Objective:To study the antibacterial activity and chemical constituents of Sojae Semen Praeparatum as a traditional Chinese medicine. Method:Sojae Semen Praeparatum was extracted with 80% ethanol,and the extracts were suspended in an appropriate amount of distilled water and extracted with petroleum ether,ethyl acetate and water saturated n-butanol in turn. The antibacterial activity in vitro was used for screening the active fractions. Using activity-guided-isolation,the active fractions were isolated and purified by silica gel column chromatography and preparative HPLC and other methods. Their structures were identified by physicochemical properties and spectroscopic analysis, such as 1H-NMR,13C-NMR and MS. Result:The ethyl acetate fraction of 80% ethanol extract from Sojae Semen Praeparatum showed a stronger activity and was studied further. Eighteen compounds were isolated, and eleven compounds were identified as daidzein (1),daidzin (2),genistein (3),genistin (4),glycitein (5),glycitin (6),daucosterol (7),thymine (8),adenine (9),uracil (10),uridine (11). Conclusion:The ethyl acetate fraction of 80% ethanol extract from Sojae Semen Praeparatum shows an antibacterial activity. Compounds 1-6 are isoflavones. Compounds 7-11 are isolated from Sojae Semen Praeparatum for the first time.

20.
China Journal of Chinese Materia Medica ; (24): 174-182, 2018.
Article in Chinese | WPRIM | ID: wpr-776406

ABSTRACT

An efficient method of ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap (UHPLC-LTQ-Orbitrap) mass spectrometer was established to elucidate the metabolites of tanshinone Ⅰ and tanshinone ⅡA in rats. Urine and plasma samples were collected after oral gavage. After processing biological sample by solid phase extraction, Waters ACQUITY HPLC BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm) was used with 0.1% formic acid (A) - acetonitrile (B) solution as the mobile phase for gradient elution. The plasma, urine and the blank samples were then analyzed by ESI-LTQ-Orbitrap equipped with an ESI ion source under positive ion mode. On the basis of the accurate mass measurements, multiple mass spectra and comparison of data with published literature, a total of 26 metabolites were tentatively identified and characterized in the rat samples. Among them, 7 metabolites were derived from tanshinone Ⅰ through metabolic pathways of glucuronide conjugation, hydroxylation, reduction reaction, demethylation reaction, methylation, sulfate conjugation and their composite reactions. Nineteen metabolites were derived from tanshinone ⅡA through metabolic pathways of hydroxylation, reduction reaction, methylation, sulfate conjugation, glucuronidation, glucosylation and their complicated reactions. The results showed that the metabolism of tanshinone Ⅰ and tanshinone ⅡA in rats could be comprehensively clarified by using UHPLC-LTQ-Orbitrap mass spectrometer, providing material basis for the further research in terms of pharmacodynamics, toxicology, and secondary development of Chinese medicine.


Subject(s)
Animals , Rats , Chromatography, High Pressure Liquid , Abietanes , Blood , Metabolism , Urine
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