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1.
Article in Chinese | WPRIM | ID: wpr-906033

ABSTRACT

Objective:To explore the potential mechanism of Qingke Pingchuan granule in treating acute and chronic bronchitis complicated with chronic obstructive pulmonary disease (COPD) by network pharmacology. Method:The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was retrieved to collect the active components of Qingke Pingchuan granule and predict the action targets, followed by the construction of component-target network using Cytoscape 3.8. GeneCards, Online Mendelian Inheritance in Man(OMIM), and DrugBank were used to harvest disease targets, whose names were put into UniProt for standardization. The treatment targets of Qingke Pingchuan Granule against the two diseases were obtained based on Venn diagram, which were then imported into the STRING platform for constructing the protein-protein interaction (PPI) network. Following the gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis based on MetaScape, the active component-common target-signaling pathway network of Qingke Pingchuan granule against acute and chronic bronchitis complicated with COPD was finally constructed. The accuracy of the target was confirmed by literature. Result:A total of 165 active components, 374 related targets, 512 disease-related targets, and 130 common targets were obtained. Among them, the 14 core therapeutic targets were further subjected to GO enrichment analysis, which yielded 390 biological processes, nine cell components, and 23 molecular functions. The KEGG pathway analysis revealed 22 signaling pathways. Conclusion:Qingke Pingchuan granule alleviates the diseases possibly by regulating such targets as vascular endothelial growth factor receptor 2(KDR), transforming growth factor beta-1 (TGF-<italic>β</italic><sub>1</sub>), caveolin 1(CAV1), hypoxia-inducible factor-1alpha(HIF-1<italic>α</italic>), and interleukin-2(IL-2), affecting the synthesis and transport of regulatory factors in cytoplasm, and controlling the cell proliferation and apoptosis.

2.
Article in English | WPRIM | ID: wpr-922110

ABSTRACT

OBJECTIVE@#To analyze clinical feature and information of medication to explore the risk signals of preparations containing Psoraleae Fructus (BGZP) related with hepatobiliary adverse drug reactions (ADR), in order to reinforce pharmacovigilance.@*METHODS@#A retrospective study was conducted based on hepatobiliary ADR related with BGZP from the China Adverse Drug Reaction Monitoring System in years from January 2012 to December 2016. Serious and general ADRs were analyzed and assessed.@*RESULTS@#There were 355 cases of hepatobiliary ADR related to BGZP. Both the amount of cases and the proportion of serious ADR showed an increasing growth by years (P<0.05). It was found that 10.43% of 355 cases may be involved with irrational drug use, including overdose, repeated medication, and combination of multiple drugs. There were 190 cases which used BGZP (non-combination), and they were mainly for common in diseases caused by abnormal immune activation (accounting for 40.53% of the total cases). Especially at the age group with the most cases with age of 41-50 years, the cases associated with immunological diseases of female were obviously more than that of male (P<0.05). The latency of hepatobiliary ADR related to BGZP ranged from 1 to 386 days, and the median latency was 27.5 days, along with the range of cumulative dose (0.45-520.02 g) as well as the daily dose (0.09-2.64 g/d) after the conversion.@*CONCLUSIONS@#Cases of hepatobiliary ADR related to BGZP showed significant individual differences, and there was no correlation between drug usage duration and dosage and the occurrence of hepatobiliary ADR. It may be similar with idiosyncratic drug-induced liver injury, and recommended that BGZP should be used with more caution under monitoring liver function, especially in female patients with immunological diseases.


Subject(s)
Adult , Adverse Drug Reaction Reporting Systems , Chemical and Drug Induced Liver Injury/etiology , Drug-Related Side Effects and Adverse Reactions , Female , Humans , Male , Middle Aged , Pharmacovigilance , Retrospective Studies
3.
Article in Chinese | WPRIM | ID: wpr-921751

ABSTRACT

The aim of this study was to investigate the mechanism of luteolin regulating lipoxygenase pathway against oxygen-glucose deprivation/reperfusion(OGD/R) injury in H9 c2 cardiomyocytes. First, Discovery Studio 2019 was used for the molecular docking of luteolin with three key enzymes including lipoxygenase 5(ALOX5), lipoxygenase 12(ALOX12), and lipoxygenase 15(ALOX15) in lipoxygenase pathway. The docking results showed that luteolin had high docking score and similar functional groups with the original ligand. From this, H9 c2 cardiomyocytes were cultured in vitro, and then the injury model of H9 c2 cardiomyocytes was induced by deprivation of oxygen-glucose for 8 h, and rehabilitation of oxygen-glucose for 12 h. Cell viability was detected by tetrazolium(MTT) colorimetry. H9 c2 cardiomyocytes were observed with a fluorescence inverted microscope, and colorimetry was used to detect the level of lactate dehydrogenase(LDH) in cell supernatant. The results showed that luteolin could significantly protect the morphology of H9 c2 cells, significantly improve the survival rate of H9 c2 cardiomyocytes in OGD/R injury model, reduce the level of LDH in cell supernatant, inhibit cytotoxicity, and maintain the integrity of cell membrane. The inflammatory cytokines interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) were detected by enzyme-linked immunosorbent assay. Compared with the model group, luteolin can significantly reduce the release of IL-6 and TNF-α. Western blot was employed to detect the protein levels of ALOX5, ALOX12, and ALOX15 in lipoxygenase pathway. After luteolin intervention, the protein levels of ALOX5, ALOX12, and ALOX15 were significantly down-regulated compared with those in model group. These results indicate that luteolin can inhibit the release of IL-6 and TNF-α by restraining the activation of lipoxygenase pathway, thereby playing a protective role in the cardiomyocyte injury model induced by OGD/R.


Subject(s)
Apoptosis , Glucose , Humans , Lipoxygenases , Luteolin/pharmacology , Molecular Docking Simulation , Myocytes, Cardiac , Oxygen , Reperfusion Injury , Signal Transduction
4.
Article in Chinese | WPRIM | ID: wpr-872653

ABSTRACT

Objective:To investigate the intervening effect of velvet antler peptide (VAP) on rotenone-induced neuroblastoma (SH-SY5Y) cell damage and explore its related mechanism. Method:0.5 μmol·L-1 rotenone was used to SH-SY5Y cells to establish an in vitro model of Parkinson's disease (PD). A blank control group, a model group, high, medium and low dose VAP groups (150,100,50 mg·L-1, respectively) and a rapamycin group were established. The number of lewy bodies, changes in mitochondrial membrane potential, content of reactive oxygen species (ROS) and α-synuclein (α-syn), protein kinase B (Akt), and mammalian target of rapamycin (mTOR) were observed by hematoxylin-eosin(HE) staining, rhodamine 123 staining, DCFH-DA staining and immunohistochemical staining expression respectively. Result:The results of HE staining showed that as compared with the blank group, the number of cells in model group was reduced, the tentacle structure became dull, the shape became round, and eosinophilic Lewy bodies were visible in cytoplasm. As compared with model group, there was no significant difference in cell morphology from rapamycin group and VAP high, medium and low dose groups, but there were fewer Lewy bodies in cytoplasm in these four groups. Rhodamine 123 staining showed that as compared with blank group, the mitochondrial membrane potential was increased significantly in model group (P<0.05). As compared with the model group, the mitochondrial membrane potential was decreased in rapamycin group and VAP high, medium and low dose groups (P<0.05). DCFH-DA staining results showed that as compared with blank group, the content of ROS was increased significantly in cells of model group (P<0.05). As compared with model group, the content of ROS was decreased in rapamycin group and VAP high, medium and low dose groups (P<0.05). Immunohistochemical staining showed that as compared with blank group, the protein expression levels of α-syn,Akt,and mTOR were increased significantly in model group (P<0.05). As compared with model group, the protein expression levels of α-syn and mTOR were significantly reduced in rapamycin group and VAP high and medium dose groups (P<0.05), and the expression levels of Akt were significantly reduced in rapamycin group and VAP high-dose group (P<0.05). Conclusion:Velvet antler peptides may play a neuroprotective role by regulating the Akt/mTOR signaling pathway and promoting the degradation of α-syn in SH-SY5Y cells.

5.
Article in Chinese | WPRIM | ID: wpr-828452

ABSTRACT

Chinese herbal medicine decoction pieces(CHMDP), one of the main forms of traditional Chinese medicine(TCM) in clinic, have been widely used. However, the irrational use is increasingly serious due to the lack of the indicators for judging the rational use of CHMDP in medical institutions and the codes and standards for the clinical use of CHMDP. In order to regulate the rational clinical use of CHMDP, improve the clinical efficacy and ensure the drug safety for the patients, clinical pharmaceutical experts and clinical medical experts from 40 third-grade class-A hospitals nationwide were organized to give the "expert consensus on clinical application of CHMDP" in terms of prescription writing, combined use of drugs, use of special drugs, and drug use for special population. Detailed analysis and argumentation were conducted in accordance with the laws and regulations, Chinese Pharmacopoeia 2015 edition, Chinese Pharmacopoeia Code Notice for Clinical Use of Medicine, Administrative Regulations for Prescriptions, Administrative Specifications for Hospital Prescription Review(interim), and Chinese Traditional Medicine Prescription Format and Writing Specifications, as well as relevant project findings.


Subject(s)
Consensus , Drugs, Chinese Herbal , Humans , Medicine, Chinese Traditional , Prescriptions , Reference Standards
6.
Article in Chinese | WPRIM | ID: wpr-777519

ABSTRACT

A retrospective study was performed in drug-induced liver injury(DILI) cases associated with Dictamni Cortex(Baixianpi,BXP) Preparations,which were treated at grade Ⅲ class A liver disease hospitals from 2008 to 2016 and spontaneously reported for adverse reactions between 2012 and 2016 at HILI Cloud(hilicloud.net). The results showed 25 DLII cases associated with BXP Preparations treated at grade Ⅲ class A liver disease hospitals during the 9 years,including only 14 cases in line with the clinical diagnostic criteria of Guidelines for the Diagnosis and Treatment of Herb-Induced Liver Injury. And 74 DILI cases associated with BXP Preparations spontaneously reports adverse reactions,and 18. 92% of them had unreasonable medication,including polypharmacy(21. 43%),overdose(28. 57%) and repeated dosage(50%). And 47 DILI cases used BXP Preparations to treat psoriasis and vitiligo(a total of59. 57%). The time range of taking BXP Preparations until liver injury occurred was 1-366 d,with the median of 18 d. The dose of BXP Preparations was estimated to be 0. 09-12 g·d-1. And the cumulative dosage of taking drugs until liver injury occurred was 1. 1-336 g. Obvious associations with time-toxicity as well as quantity-toxicity could not be found based on the wide range of time-toxicity relations and quantity-toxicity relations. On the basis of the study,we found that DILI cases associated with BXP Preparations commonly occurred in patients with immune diseases,such as psoriasis and vitiligo,indicating specific individual differences. The results suggested that DILI cases associated with BXP Preparations would be correlated with the property of idiosyncratic drug-induced liver injury. In conclusion,the risk of liver injury clinically caused by BXP Preparations should be paid more attention,and the studies on the mechanism of idiosyncratic drug-induced liver injury must be enhanced,and those on risk factors,like irrational drug use,should be strengthened. Moreover,the evaluation of the risk-to-benefit ratio is supposed to be performed for the sake of improving the risk prevention and control standards for BXP preparations,and ensuring safe and rational clinical application of BXP Preparations.


Subject(s)
Chemical and Drug Induced Liver Injury , Epidemiology , China , Dictamnus , Chemistry , Drugs, Chinese Herbal , Humans , Liver , Retrospective Studies , Risk Factors
7.
Chinese Pharmaceutical Journal ; (24): 608-613, 2018.
Article in Chinese | WPRIM | ID: wpr-858363

ABSTRACT

OBJECTIVE: To compare the differences of macrophages phagocytic activities between Dioscorea opposita Thunb. cv. Tiegun(abbreviated as TG) and Dioscorea opposita Thunb. but not Tiegun(abbreviated as NTG). METHODS: CCK-8 assay was used to test the cytotoxicity. On the basis of non-toxic dose, the high content screening(HCS) cell imaging analysis was applied to test the phagocytic ability of RAW264.7 cells engulfing GFP-E. coli in vitro, and the phagocytic rates between different kinds of samples at various concentrations were calculated. Furthermore, the biological potency was measured according to the qualitative response parallel method to better evaluate the difference of TG and NTG, by translating phagocytic rates into biological potency. RESULTS: At the range of 0.695 - 5.56 mg(crude drug) •mL-1, all 11 batches samples have no toxicity. The results of HCS displayed that every sample could promote cell phagocytic activity in varying degrees at a certain concentration. RAW264.7 cells could engulf the GFP-E. coli, and the images of HCS reflected the situation of phagocytosis clearly. In addition, the results of biopotency showed that the biopotency value of different samples was between 39.56 to 100, and the average biopotency value of TG was significantly higher than NTG (P < 0.05). CONCLUSION: In vitro experiments showed that all the samples could promote the phagocytic activity of RAW264.7 cells at different degrees. And the average biopotency value of TG was significantly higher than NTG, which is consistent with the saying that "TG has a better quality".

8.
Article in Chinese | WPRIM | ID: wpr-230970

ABSTRACT

Chinese medicine prescriptions are a type of medical documents written by doctors after they understand the patients' conditions for syndrome differentiation. Chinese medicine prescriptions are also the basis for pharmacy personnel to dispense medicines and guide patients to use drugs. It has the legal, technical and economic significances. Chinese medicine prescriptions contain such information of names, quantity and usage. Whether the names of drugs in Chinese medicine prescriptions are standardized or not is directly related to the safety and efficacy of the drugs. At present, nonstandard clinical prescriptions are frequently seen. With "Chinese medicine prescription", "names of drug in Chinese medicine prescription" and "standards of Chinese medicine prescription" as key words, the author searched CNKI, Wanfang and other databases, and consulted nearly 100 literatures, so as to summarize current names of drugs in traditional Chinese medicine prescription, analyze the reasons, and give suggestions, in the expectation of standardizing the names of drugs used in traditional Chinese medicine prescriptions.

9.
Chinese Medical Journal ; (24): 723-730, 2016.
Article in English | WPRIM | ID: wpr-328168

ABSTRACT

<p><b>BACKGROUND</b>The aim of this study was to assess the efficacy and safety of vinorelbine and cisplatin (NP chemotherapy) alone or in combination with Aidi injection for the treatment of advanced nonsmall cell lung cancer (NSCLC).</p><p><b>METHODS</b>Pertinent publications were identified in PubMed, EMBASE, Cochrane Library, CNKI, CQVIP, and Wanfang databases, up to December 8, 2015. After quality assessment of all included randomized controlled trials evaluating Aidi injection combined with NP chemotherapy for the treatment of advanced NSCLC, a meta-analysis was performed by Review Manager 5.2 and STATA 12.0 for statistical analyses.</p><p><b>RESULTS</b>Twelve studies including 509 and 503 cases in the experimental and control groups, respectively, were finally analyzed. The meta-analysis revealed that when cisplatin dose ranging from 20 to 40 mg/m 2 , combination of Aidi injection and NP chemotherapy was statistically different compared with NP chemotherapy alone in enhancing efficiency (relative risk [RR] = 1.24, 95% confidence interval [CI] [1.05-1.47], P = 0.010) and reducing the incidence of Grade II or above nausea and vomiting (RR = 0.49, 95% CI [0.30-0.80], P = 0.005). Meanwhile, with cisplatin ranging from 80 to 120 mg/m 2 , no significant differences in efficiency (RR = 1.11, 95% CI [0.87-1.42], P = 0.390) and Grade II or above nausea and vomiting (RR = 0.88, 95% CI [0.71-1.10], P = 0.260) were obtained. In addition, Aidi injection combined with NP chemotherapy was superior to NP chemotherapy alone in improving the quality of life, alleviating Grade II or above leukopenia and thrombocytopenia.</p><p><b>CONCLUSIONS</b>Aidi injection combined with NP chemotherapy can enhance efficiency, improve the quality of life, and decrease adverse effects in patients with advanced NSCLC.</p>


Subject(s)
Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Carcinoma, Non-Small-Cell Lung , Drug Therapy , Psychology , Cisplatin , Drugs, Chinese Herbal , Humans , Injections , Lung Neoplasms , Drug Therapy , Psychology , Publication Bias , Quality of Life , Vinblastine
10.
Article in Chinese | WPRIM | ID: wpr-246165

ABSTRACT

The commodity specification and grade of Chinese medicinal materials is a measure of the quality of traditional Chinese medicines (TCMs), which directly impacts on the safety and effectiveness of clinical medicines. It is an urgent problem to establish a set of standards which can both interpret the scientific connotation of the commodity specification and grade of Chinese medicinal materials and play a significant role on clinical medicines as well as markets. This paper reviews the research methods of the commodity specification and grade of Chinese medicinal materials such as sensory evaluation, chemical assessment, biological evaluation, and cited the applications of various methods for the classification of TCMs. It provides technical support for establishing standards of the commodity specification and grade of Chinese medicinal materials, and also constructs scientific basis for clinical rational drug use.


Subject(s)
Drugs, Chinese Herbal , Chemistry , Economics , Humans , Medicine, Chinese Traditional , Economics , Reference Standards , Plants, Medicinal , Chemistry , Quality Control , Research Design
11.
Chinese Pharmaceutical Journal ; (24): 1222-1225, 2014.
Article in Chinese | WPRIM | ID: wpr-859923

ABSTRACT

METHODS: Female Kunming mice were randomly divided into five groups, ie, blank control group, low-dose group, middle-dose group, high-dose group, and the positive control group. The three treatment groups were given various doses of Rhodiola granules, ie, 0. 1, 1, 3 g��kg-1 respectively, the positive control group were given Nuodikang capsules at 280 mg��kg-1, and the blank control group was given deionized water for 35 d. The levels of SOD, MDA, GSH-PX in serum, liver, and brain were determined. For the experiment in vitro, with vitamin C as the control drug, the absorbance of Rhodiola granule solution at concentrations of 0.01, 0.1, 1, 10 mg��mL-1 was measured, and the activity of scavenging superoxide anion free radicals and hydroxyl free radicals was calculated.

12.
Article in Chinese | WPRIM | ID: wpr-642715

ABSTRACT

Objective To determine effects of T-2 toxin and selenium on expression of aggrecanase in human chondrocyte.Methods Human chondrocytes were treated with T-2 toxin(0,1,10,20 μg/L),and/or sodium selenite(final concentration of selenium 0,0.1 mg/L) for 5 days.Aggrecan expression was determined by Western blotting,aggrecanase-1 and aggrecanase-2 mRNA levels were measured by RT-PCR.ResultsSelenium and T-2 toxin had effects on both aggrecan protein levels and its aggrecanases(include aggrecanase-1 and aggrecanase-2 ) mRNA levels(F =0.294,27.71 for aggrecan,F =107.45,362.25 for aggrecanase-l,F =34.68,22.26 for aggrecanase-2,respectively,all P < 0.05),and there was interaction between selenium and T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression(F =79.99,230.76,388.33,all P < 0.05).Furthermore,selenium presented significant antagonism to T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression.Aggrecan expression levels(0.278 ± 0.015,0.235 ± 0.029,0.195 ± 0.028,0.399 ± 0.028,0.299 ± 0.020,0.263 ±0.019) induced by both 1,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium were significantly decreased than the levels(0.472 ± 0.0358,0.197 ± 0.018,all P < 0.05) in control group(0 mg/L toxin).Selenium partially blocked the effects induced by 1,10,and 20 μg/L T-2 toxin(all P< 0.05).One,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium increased both aggrecanase-1 mRNA levels(0.535 ± 0.033,1.071 ± 0.043,1.454 ± 0.058,1.057 ±0.048,0.555 ± 0.036,0.902 ± 0.045) and aggrecanase-2 mRNA levels(0.596 ± 0.038,0.656 ± 0.033,0.949 ±0.049,0.600 ± 0.040,0.453 ± 0.031,0.164 ± 0.011),when compared with control(0.481 ± 0.023,0.346 ±0.020 for aggrecanase-1 and 0.387 ± 0.020,1.021 ± 0.046 for aggrecanase-2,respectively,all P < 0.05).Selenium partially blocked 10,20 μg/L T-2 toxins induced upregulation of aggrecanase-1 (all P < 0.05) and aggrecanase-2 (all P < 0.05 ).Conclusions These data suggest a possible molecular mechanism that T-2 toxin could induce cartilage matrix degradation through the upregulation of aggrecanases expression and enzyme activities.Trace element selenium has some protective effect on cartilage proteoglycan degradation induced by T-2 toxins.

13.
Article in Chinese | WPRIM | ID: wpr-325158

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the use of cancellous bone matrix gelatin (BMG) combined with chondrocytes in constructing tissue-engineered cartilage by observing the growth, proliferation and differentiation of chondrocytes on allogeneic cancellous BMG.</p><p><b>METHODS</b>The articular chondrocytes isolated from a 1-month-old rabbit were multiplied to a monolayer and seeded onto cancellous BMG to construct tissue-engineered cartilage in vitro during a period of 6 weeks. Samples were taken from the construct after 1, 2, 4, and 6 weeks of culture and evaluated by histology, immunohistochemistry and transmission electron microscopy (TEM).</p><p><b>RESULTS</b>The chondrocytes excreted matrix proteoglycan and collagen on cancellous BMG. With the prolongation of the culture time, the cells proliferated in the construct and the cells in the lacunae increased. Numerous chondrocytes were present the central region of the cancellous BMG and surrounded by extracellular matrix. By 6 weeks of culture, the BMG was covered with 15-20 layers of chondrocytes and cartilaginous tissue occurred in the pores throughout the cancellous BMG. Immunohistochemical staining showed rich and evenly distributed type II collagen around the chondrocytes, and TEM revealed an ultrastructure of the chondrocyte similar to that of native chondroctyes, with abundant extracellular matrix produced around the cells.</p><p><b>CONCLUSION</b>Tissue-engineered cartilage can be constructed in vitro using allogeneic cancellous BMG combined with chondrocytes. Allogeneic cancellous BMG serves as a good scaffold material for tissue-engineered cartilage to promote the growth and proliferation of the seeded chondrocytes and allows maintenance of the differentiation phenotype of the cells.</p>


Subject(s)
Absorbable Implants , Animals , Bone Matrix , Chemistry , Cartilage , Cell Biology , Cells, Cultured , Chondrocytes , Cell Biology , Physiology , Gelatin , Chemistry , Rabbits , Tissue Engineering , Methods , Tissue Scaffolds
14.
Article in Chinese | WPRIM | ID: wpr-315181

ABSTRACT

<p><b>OBJECTIVE</b>To establish a new method for screening active ingredients of Chinese herbs by determining different bio-thermodynamic effects of 3 genosides on splenic lymphocyte of mice.</p><p><b>METHODS</b>Using a thermal bioactivity monitoring system, the maximum heat output (mHO), average metabolic heat (MH) and constant of decrease rate (DR) of lymphocyte were determined based on the growth metabolic power-time curve, and the outcomes were verified by MIT.</p><p><b>RESULTS</b>The mHO and MH increased and the DR decreased after lymphocytes being exposed to the 3 genosides in different concentrations, arranged upon their potency as genoside Rg3 > genoside Rg2 > genoside Rg1 (merely insignificant effect). MTT showed the same results.</p><p><b>CONCLUSION</b>Heat activity monitoring system could precisely display the different bio-thermal dynamic effects of 3 genosides on splenic lymphocyte.</p>


Subject(s)
Animals , Cell Survival , Cells, Cultured , Energy Metabolism , Ginsenosides , Pharmacology , Lymphocytes , Cell Biology , Metabolism , Male , Mice , Mice, Inbred BALB C , Spleen , Cell Biology , Thermodynamics
15.
Article in English | WPRIM | ID: wpr-359406

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of T-2 toxin on expressions of Fas, p53, Bcl-xL, Bcl-2, Bax and caspase-3 on human chondrocytes.</p><p><b>METHODS</b>Human chondrocytes were treated with T-2 toxin (1-20 ng/ml) for 5 d. Fas, p53 and other apoptosis-related proteins such as Bax, Bcl-2, Bcl-xL, caspase-3 were determined by Western blot analysis and their mRNA expressions were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Increases in Fas, p53 and the pro-apoptotic factor Bax protein and mRNA expressions and a decrease of the anti-apoptotic factor Bcl-xL were observed in a dose-dependent manner after exposures to 1-20 ng/ml T-2 toxin, while the expression of the anti-apoptotic factor Bcl-2 was unchanged. Meanwhile, T-2 toxin could also up-regulate the expressions of both pro-caspase-3 and caspase-3 in a dose-dependent manner.</p><p><b>CONCLUSION</b>These data suggest a possible underlying molecular mechanism for T-2 toxin to induce the apoptosis signaling pathway in human chondrocytes by regulation of apoptosis-related proteins.</p>


Subject(s)
Apoptosis , Base Sequence , Blotting, Western , Caspase 3 , Genetics , Metabolism , Cell Proliferation , Cells, Cultured , Chondrocytes , Cell Biology , Metabolism , DNA Primers , Genetics , Gene Expression , Humans , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , T-2 Toxin , Toxicity , Tumor Suppressor Protein p53 , Genetics , Metabolism , bcl-2-Associated X Protein , Genetics , Metabolism , bcl-X Protein , Genetics , Metabolism , fas Receptor , Genetics , Metabolism
16.
Article in English | WPRIM | ID: wpr-277315

ABSTRACT

<p><b>OBJECTIVE</b>To identify the relationship between T-2 toxin and Kashin-Beck disease (KBD), the effects of T-2 toxin on aggrecan metabolism in human chondrocytes and cartilage were investigated in vitro.</p><p><b>METHODS</b>Chondrocytes were isolated from human articular cartilage and cultured in vitro. Hyaluronic acid (HA), soluble CD44 (sCD44), IL-1beta and TNF-alpha levels in supernatants were measured by enzyme-linked immunosorbent assay (ELISA). CD44 content in chondrocyte membrane was determined by flow cytometry (FCM). CD44, hyaluronic acid synthetase-2 (HAS-2) and aggrecanases mRNA levels in chondrocytes were determined using reverse transcription polymerase chain reaction (RT-PCR). Immunocytochemical method was used to investigate expressions of BC-13, 3-B-3(-) and 2-B-6 epitopes in the cartilage reconstructed in vitro.</p><p><b>RESULTS</b>T-2 toxin inhibited CD44, HAS-2, and aggrecan mRNA expressions, but promoted aggrecanase-2 mRNA expression. Meanwhile, CD44 expression was found to be the lowest in the chondrocytes cultured with T-2 toxin and the highest in control plus selenium group. In addition, ELISA results indicated that there were higher sCD44, IL-1beta and TNF-alpha levels in T-2 toxin group. Similarly, higher HA levels were also observed in T-2 toxin group using radioimmunoprecipitation assay (RIPA). Furthermore, using monoclonal antibodies BC-13, 3-B-3 and 2-B-6, strong positive immunostaining was found in the reconstructed cartilage cultured with T-2 toxin, whereas no positive staining or very weak staining was observed in the cartilage cultured without T-2 toxin. Selenium could partly inhibit the effects of T-2 toxin above.</p><p><b>CONCLUSION</b>T-2 toxin could inhibit aggrecan synthesis, promote aggrecanases and pro-inflammatory cytokines production, and consequently induce aggrecan degradation in chondrocytes. These will perturb metabolism balance between aggrecan synthesis and degradation in cartilage, inducing aggrecan loss in the end, which may be the initiation of the cartilage degradation.</p>


Subject(s)
Cartilage, Articular , Metabolism , Cells, Cultured , DNA , Flow Cytometry , Humans , Hyaluronan Receptors , Immunohistochemistry , Interleukin-1beta , Proteoglycans , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Selenium , Pharmacology , T-2 Toxin , Toxicity , Tumor Necrosis Factor-alpha
17.
Article in Chinese | WPRIM | ID: wpr-268119

ABSTRACT

<p><b>OBJECTIVE</b>To observe cell apoptosis and Bcl-2 and Bax expression changes of chondrocytes induced by butenolide (BUT) and the inhibitory effect of selenium against BUT-induced chondrcyte apoptosis, to gain insights into the mechanism by which BUT induces chondrcyte apoptosis.</p><p><b>METHODS</b>Cartilage tissue reestablished from human fetal articular chondrocytes in vitro were treated with BUT at the concentrations of 0.1, 1.0 and 5.0 microg/ml and with the protective factor selenium. TUNEL method was used to detect chondrocyte apoptosis, which was quantified by flow cytometry. Immunohitochemistry was performed to analyze the expression of Bcl-2 and Bax in the reestablished cartilage tissue.</p><p><b>RESULTS</b>BUT exposure induced chondrocyte apoptosis, and the apoptosis rate increased with the concentration increment of BUT from 0 to 1.0 mg/ml, resulting also increased positive expression rate of Bcl-2 and Bax(P<0.05). The apoptosis rate of chondrocytes in BUT+ selenium group was significantly lower than that of BUT groups (P<0.05), as was the positivity rate of Bcl-2 and Bax expression (P<0.05).</p><p><b>CONCLUSION</b>BUT induces chondrocyte apoptosis in positive relation with BUT concentration (from 0 to 1.0 mg/ml) and causes increased expressions of Bcl-2 and Bax. Selenium can inhibit the chondrocyte apoptosis induced by BUT.</p>


Subject(s)
4-Butyrolactone , Pharmacology , Apoptosis , Cells, Cultured , Chondrocytes , Humans , In Situ Nick-End Labeling , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Selenium , Pharmacology , bcl-2-Associated X Protein , Metabolism
18.
Chinese Journal of Traumatology ; (6): 266-271, 2006.
Article in English | WPRIM | ID: wpr-280899

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of cancellous bone matrix gelatin (BMG) engineered with allogeneic chondrocytes in repairing articular cartilage defects in rabbits.</p><p><b>METHODS</b>Chondrocytes were seeded onto three-dimensional cancellous BMG and cultured in vitro for 12 days to prepare BMG-chondrocyte complexes. Under anesthesia with 2.5% pentobarbital sodium (1 ml/kg body weight), articular cartilage defects were made on the right knee joints of 38 healthy New Zealand white rabbits (regardless of sex, aged 4-5 months and weighing 2.5-3 kg) and the defects were then treated with 2.5% trypsin. Then BMG-chondrocyte complex (Group A, n=18), BMG (Group B, n=10), and nothing (Group C, n=10) were implanted into the cartilage defects, respectively. The repairing effects were assessed by macroscopic, histologic, transmission electron microscopic (TEM) observation, immunohistochemical examination and in situ hybridization detection, respectively, at 2, 4, 8, 12 and 24 weeks after operation.</p><p><b>RESULTS</b>Cancellous BMG was degraded within 8 weeks after operation. In Group A, lymphocyte infiltration was observed around the graft. At 24 weeks after operation, the cartilage defects were repaired by cartilage tissues and the articular cartilage and subchondral bone were soundly healed. Proteoglycan and type II collagen were detected in the matrix of the repaired tissues by Safranin-O staining and immunohistochemical staining, respectively. In situ hybridization proved gene expression of type II collagen in the cytoplasm of chondrocytes in the repaired tissues. TEM observation showed that chondrocytes and cartilage matrix in repaired tissues were almost same as those in the normal articular cartilage. In Group B, the defects were repaired by cartilage-fibrous tissues. In Group C, the defects were repaired only by fibrous tissues.</p><p><b>CONCLUSIONS</b>Cancellous BMG can be regarded as the natural cell scaffolds for cartilage tissue engineering. Articular cartilage defects can be repaired by cancellous BMG engineered with allogeneic chondrocytes. The nature of repaired tissues is closest to the normal cartilage. Local administration of trypsin can promote the adherence of repaired tissues to host tissues. Transplantation of allogeneic chondrocytes has immunogenicity, but the immune reaction is weak.</p>


Subject(s)
Animals , Bone Matrix , Cartilage, Articular , Pathology , General Surgery , Cells, Cultured , Chondrocytes , Allergy and Immunology , Transplantation , Collagen Type II , Female , Gelatin , Immunohistochemistry , Male , Rabbits , Tissue Engineering , Methods
19.
Article in Chinese | WPRIM | ID: wpr-255305

ABSTRACT

<p><b>OBJECTIVE</b>To study the inhibitory effect of T-2 toxin on the expression of aggrecan and collagen II in chondrocytes and the protection of selenium against this effect.</p><p><b>METHODS</b>Human chondrocytes cultured in vitro were treated with T-2 toxin at different concentrations for varied time periods (1-5 days), and the cell viability was measured by MTT assay. Aggrecan expression was detected by toluidine blue staining and collagen II expression by immunostaining using monoclonal antibody of collagen. Aggrecan and collagen II mRNA expressions were measured by semiquantitative RT-PCR.</p><p><b>RESULTS</b>T-2 toxin dose- and time-dependently affected chondrocyte viability within the concentration range of 0.001-2 mg/L, the prolonged treatment time further enhanced the dose dependence of the inhibitory effect. T-2 toxin lowered aggrecan and collagen II synthesis in the chondrocytes and reduced their mRNA expressions. Selenium could partly attenuate the inhibitory effects of T-2 toxin on aggrecan mRNA expression, but showed no such effect against T-2-induced collagen II expression.</p><p><b>CONCLUSION</b>T-2 toxin can obviously inhibit aggrecan and collagen II synthesis in human chondrocytes, and selenium can partly antagonize the inhibitory effects of T-2 toxin on aggrecan.</p>


Subject(s)
Aggrecans , Genetics , Cells, Cultured , Chondrocytes , Cell Biology , Metabolism , Collagen Type II , Genetics , Dose-Response Relationship, Drug , Fetus , Humans , Protective Agents , Pharmacology , RNA, Messenger , Genetics , Selenium , Pharmacology , T-2 Toxin , Toxicity
20.
Article in Chinese | WPRIM | ID: wpr-281192

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of butenolide (BUT) on cultured chondrocytes differentiation and the possible protective effects of selenium (Se).</p><p><b>METHODS</b>Ex-vivo cultured chondrocytes were divided into six groups: (1) Control group (without BUT and Se); (2) Se 0.1 microg/ml control group; (3) BUT 0.1 microg/ml group; (4) BUT 1.0 microg/ml group; (5) BUT 5.0 microg/ml group; and (6) BUT 1.0 microg/ml + Se 0.1 microg/ml group. The expression of collagen II (Col II), collagen X (ColX), basic fibroblast growth factor (bFGF), and parathyroid hormone-related peptide (PTHrP) in (or around) chondrocytes in all groups were analyzed by immunohistochemistry.</p><p><b>RESULTS</b>The expressions of Col II in 1.0 microg/ml BUT group and 5.0 microg/ml BUT group were significantly lower than those in the control group (P < 0.05). The expression of Col II in 1.0 microg/ml BUT + Se group were significantly higher than those in the 1.0 microg/ml BUT group and 5.0 microg/ml BUT group (P < 0.05). The expressions of bFGF and PTHrP of BUT groups were significantly higher than those in the Se and control groups (P < 0.05). No expression of ColX was observed in all groups.</p><p><b>CONCLUSION</b>BUT can affect the collagen II synthesis of the chondrocytes. Selenium supplementation may play a protective role.</p>


Subject(s)
4-Butyrolactone , Pharmacology , Cell Differentiation , Cells, Cultured , Chondrocytes , Cell Biology , Humans , Protective Agents , Pharmacology , Selenium , Pharmacology , T-2 Toxin , Toxicity
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