ABSTRACT
Polysaccharide is among the main active components of Ganoderma lucidum for tumor prevention and treatment. Howe-ver, it remains unclear whether it has synergy with tumor immunotherapy. This study evaluated the effect of G. lucidum polysaccharides(GLP) on the infiltration of T lymphocytes into tumor and the underlying mechanism, in order to provide a reference for its application in tumor immunotherapy. GLP were prepared by water extraction and alcohol precipitation combined with Sevag method and then given(intraperitoneal injection) to the mice bearing B16-F10 cells at 25, 50 and 100 mg kg~(-1), respectively, to evaluate the effect on tumor growth. The infiltration of CD3~+ and CD8~+ T cells and the expression of intercellular cell adhesion molecule-1(ICAM-1) in tumor were detected by immunohistochemistry. EA.hy926 cells were treated with 50, 100 and 200 μg·mL~(-1) GLP, and the expression of ICAM-1 was determined by Western blot. The adhesion of EA.hy926 cells treated with GLP was measured with fluorescence-labeled Jurkat cells. To analyze the mechanism based on NF-κB pathway, this study determined the protein levels of nuclear factor kappa-B(NF-κB) p65, alpha inhibitor of NF-κB(IκBα), p-NF-κB p65 and p-IκBα by Western blot. The results showed that GLP can significantly inhibit the tumor growth in mice bearing B16-F10 cells, promote the infiltration of CD3~+ and CD8~+ T cells in tumor, and increase the expression of ICAM-1 in tumor. Meanwhile, GLP could also enhance the expression of ICAM-1 in EA.hy926 cells, thus strengthen the adhesion to Jurkat cells, induce phosphorylation and protein degradation of IκBα, and raise the expression and phosphorylation level of NF-κB p65. These results suggested that GLP could promote the expression of ICAM-1 through NF-κB pathway and further enhance the infiltration of T lymphocytes into tumor, thereby inhibiting tumor growth. This study lays a foundation for the further application of GLP in tumor immunotherapy.
Subject(s)
Animals , Mice , Endothelial Cells/metabolism , Intercellular Adhesion Molecule-1/genetics , NF-kappa B/metabolism , Neoplasms , Polysaccharides , Reishi , Signal Transduction , T-Lymphocytes , Tumor Necrosis Factor-alphaABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of IPS-B2 on mouse peritoneal macrophages and the transcription of IL-1beta, IL-6, TNF-alpha and iNOS.</p><p><b>METHOD</b>ELISA method and Griess method were used to detect the effect of mouse peritoneal macrophages produce cytokines IL-1beta, IL-6, TNF-alpha and cytotoxic effectors NO. The transcription of IL-1beta, IL-6, TNF-alpha and iNOS was detected by real-time RT-PCR method.</p><p><b>RESULT</b>IPS-B2 could not promote mouse peritoneal macrophage production, but it could significantly improve the IL-1beta, IL-6, TNF-alpha content in mouse peritoneal macrophages culture supernatant, and increase the gene expression of IL-1beta, IL-6, TNF-alpha and iNOS.</p><p><b>CONCLUSION</b>IPS-B2 can enhance the ability of peritoneal macrophages to excrete bioactive substances and promote the transcription of bioactive substances to antitumor.</p>
Subject(s)
Animals , Mice , Agaricales , Chemistry , Gene Expression Regulation , Interleukin-1beta , Genetics , Interleukin-6 , Genetics , Macrophages, Peritoneal , Metabolism , Nitric Oxide , Nitric Oxide Synthase Type II , Metabolism , Polymerase Chain Reaction , Polysaccharides , Pharmacology , RNA, Messenger , Transcription, Genetic , Tumor Necrosis Factor-alpha , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the metabolic regulation and apoptosis of Sailong bone extracts on rat osteoblast cells in vitro.</p><p><b>METHOD</b>Sailong bone fat-soluble extract, Sailong bone ethanol extract and Sailong bone aqueous extract were extracted with super critical fluid extraction (SCFE) , and Sailong bone boiling water component was extracted with distilled water directly. MTT assay was applied to determine the proliferation of the cell promoted by four Sailong bone extracts and PAS assay for the aqueous proportion of the cell at different doses.</p><p><b>RESULT</b>Sailong bone fat-soluble and aqueous extract (each 10 mg x mL (-1)) could significantly improve the proliferation of rat osteoblast cells ROS 17/2. 8 (P < 0. 01). Compared with the blank, the proportion of xub-G, of the different extracts from Sailong bone is reduce evidently. The result have shown the extracts from Sailong bone could reduce the rate of aqueous of cell and could suspend the aqueous.</p><p><b>CONCLUSION</b>Sailong bone can promoting the proliferation, degrading the rate of the apoptosis and delay the development of osteoblast to be the substitute of the bone of tiger as a Chinese materia medica.</p>
Subject(s)
Animals , Rats , Apoptosis , Bone and Bones , Chemistry , Cell Proliferation , Cells, Cultured , Dose-Response Relationship, Drug , Materia Medica , Pharmacology , Osteoblasts , Cell Biology , Rodentia , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To study the effect of Spatholobus suberectus on proliferation and the hematonic mechanism.</p><p><b>METHOD</b>The techniques of culture of hematopoietic cell and hematopoietic growth factor (HGF) assay were used.</p><p><b>RESULT</b>Spatholobus suberectus could obviously promote the proliferation of bone marrow cells in healthy and anaemic mice. The culture media of spleen cell, macrophage, lung and skeletal muscle treated with S. suberectus had much stronger stimulating effects on hematopoietic cells.</p><p><b>CONCLUSION</b>S. suberectus may enhance hematopoiesis by directly or indirectly stimulating stroma cell in hematopoietic inductive microenvironment and muscle tissue to secrete some HGF (Epo, GM-CSF, IL, and MK-CSF). This is one of the biological mechanisms for hematonic effect of S. suberectus.</p>