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1.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 592-596, 2016.
Article in Chinese | WPRIM | ID: wpr-328255

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of Modified Hangqi Chifeng Decoction (MHCD) on levels of collagen type IV (Col IV), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2) in extracellular matrix (ECM) of glomerular mesangial cells (GMCs) in LPS induced mice.</p><p><b>METHODS</b>Normal serum and telmisartan, high, medium, low dose MHCD containing serums were prepared by using serum pharmacology method. GMCs were cultured in vitro. The proliferation of mesangial cells were induced using LPS as stimulating factor. GMCs were divided into six groups, i.e., the normal group, the model group, the telmisartan group, high, medium and low dose MHCD groups. Col IV content in the supernatant of mesangial cells was detected using ELISA. Protein expressions of MMP-2 and TIMP-2 were detected using Western blot.</p><p><b>RESULTS</b>Compared with the normal group, Col IV content obviously increased in the model group after 72-h LPS stimulation; protein expressions of MMP-2 and TIMP-2 were obviously up-regulated, and MMP-2/TIMP-2 ratio was down-regulated in the model group (P < 0.01). Compared with the model group, Col IV content obviously decreased in high and medium dose MHCD groups and the telmisartan group (P < 0.01); protein expressions of MMP-2 were obviously down-regulated in medium and low dose MHCD groups (P < 0.01, P < 0.05); the protein expression of TIMP-2 was obviously down-regulated in high, medium, low dose MHCD groups and the telmisartan group (P < 0.01). The pro- tein expression of TIMP-2 was obviously lower in the high dose MHCD group than in the low dose MHCD group (P < 0.01). MMP-2/TIMP-2 ratio was obviously up-regulated in the telmisartan group, high and medium dose MHCD groups (P < 0.01).</p><p><b>CONCLUSION</b>MHCD could regulate disordered MMP-2/TIMP-2 ratio in LPS induced ECM, inhibit excessive production of Col IV in ECM, promote the degradation of ECM, reduce the accumulation of ECM, thereby, delaying the process of glomerular sclerosis.</p>


Subject(s)
Animals , Mice , Cells, Cultured , Collagen Type IV , Metabolism , Extracellular Matrix , Metabolism , Kidney Glomerulus , Cell Biology , Matrix Metalloproteinase 2 , Metabolism , Mesangial Cells , RNA, Messenger , Metabolism , Tissue Inhibitor of Metalloproteinase-2 , Metabolism
2.
Acta Pharmaceutica Sinica ; (12): 1117-1123, 2014.
Article in Chinese | WPRIM | ID: wpr-299159

ABSTRACT

This study aimed to examine whether ophiopogonin D (OP-D) is capable of protecting cardiomyocytes against DOX-induced injury and the mechanisms involved. H9c2 cells were cultured. MTT assay was used to evaluate cell viability and toxicity. Mito-tracker as fluorescence probe was used to measure ROS content raised from mitochondria. The mRNA and protein expression of ATF6alpha, GRP78 and CHOP were analyzed using real-time PCR and Western blotting, respectively. The results showed that a significant endoplasmic reticulum stress (ERS) was induced upon exposure of H9c2 cells to DOX as indicated by the increase in the expression of ERS related proteins, which was paralleled with the accumulation of reactive oxygen species (ROS) and decrease in the viability of H9c2 cells. Whereas, DOX-induced ROS accumulation and up-regulation of ERS related proteins were partially abolished by pretreatment with OP-D. Consequently, a DOX-induced ERS was mitigated by application of OP-D. Similarly, DOX-induced decrease in cell viability was partially attenuated by either inhibiting CHOP or pretreatment with N-acetylcysteine (NAC), an antioxidant. Moreover, cardiac ultrastructural abnormalities seen in mouse receiving DOX injections were obviously ameliorated by pretreatment of OP-D. Taken together, the present study proved that OP-D protects cardiomyocytes against DOX-induced injury, at least in part, through reducing ROS accumulation and alleviating ERS.


Subject(s)
Animals , Mice , Rats , Acetylcysteine , Activating Transcription Factor 6 , Metabolism , Antioxidants , Cell Line , Cell Survival , Doxorubicin , Endoplasmic Reticulum Stress , Heat-Shock Proteins , Metabolism , Mitochondria , Metabolism , Myocytes, Cardiac , Reactive Oxygen Species , Metabolism , Saponins , Pharmacology , Spirostans , Pharmacology , Transcription Factor CHOP , Metabolism , Up-Regulation
3.
Journal of Southern Medical University ; (12): 2277-2280, 2010.
Article in Chinese | WPRIM | ID: wpr-323683

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the changes in white blood cell populations, lymphocyte subsets and stress-related cytokines after long-term exercise training and address the association between blood cell changes and stress-related cytokines in relation to exercise.</p><p><b>METHODS</b>A total of 1038 professional athletes were examined for CBC with Sysmex XE2100, and the T, B, and NK lymphocyte subsets were analyzed with flow cytometry. The testees' RNA were extracted from 1 ml whole blood, and the stress-related cytokines such as CRP, SELL,TNF-α, IL8, IL4, ICAM1, PECAM1, IL6, and NOS were tested by multi-RT-PCR and fragments separated by capillary electrophoresis using Beckman Coulter GeXP system.</p><p><b>RESULTS</b>No obvious difference was found in WBC count between the athletes, all within normal range. The proportion of lymphocytes was increased in the athletes by 20%-40% in comparison with the normal level, and the CD3+, CD3+CD4+, and CD3+CD8+ T, B, and NK lymphocyte subsets were all lower in the athletes than the normal range. The cytokine expressions exhibited no significant gender-related difference. IL-8, TNF-α and SELL expressions increased while IL-4 decreased in the athletes. Correlations were noted between the changes of the cells and the cytokine expressions.</p><p><b>CONCLUSION</b>Long-term exercise training affects the immune system and cause stress, which may potentially increase the risks of some chronic diseases.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Athletes , Cytokines , Physiology , Exercise , Physiology , Flow Cytometry , Immunologic Factors , Physiology , Lymphocyte Count , Lymphocyte Subsets , Cell Biology , Allergy and Immunology , Physiology , Stress, Physiological , Allergy and Immunology
4.
Biomedical and Environmental Sciences ; (12): 237-243, 2009.
Article in English | WPRIM | ID: wpr-360670

ABSTRACT

<p><b>OBJECTIVE</b>To prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin (IgY) to build an enzyme-linked immunosorbent assay (ELISA) for citrinin (CTN).</p><p><b>METHODS</b>CTN was conjugated with bovine serum albumin (BSA), ovalbumin (OVA) with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet (UV) spectrometry and Infrared (IR) spectrometry. Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). ELISA with IgY was established. Cross-reactivity of IgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied.</p><p><b>RESULTS</b>UV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA. SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD. The indirect competitive ELISA showed that the detection limit of CTN was 10 ng x mL(-1), with a good linearity ranging 20-640 ng x mL(-1).</p><p><b>CONCLUSION</b>Artificial antigens of CTN can be successfully synthesized. The established ELISA can be used to determine CTN- contaminated samples.</p>


Subject(s)
Animals , Female , Antibody Specificity , Antigens , Chemistry , Chickens , Citrinin , Chemistry , Egg Yolk , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Methods , Immunoglobulins , Allergy and Immunology
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