1.Effects of proteasome inhibitor PS-341 on the multiple cytokine expressions of mesenchymal stem cells from bone marrow in patients with multiple myeloma.
Ru-Feng LIN ; Hua LU ; Peng LIU ; Wen-Yi SHEN ; Jian-Fu ZHANG ; Yu-Jie WU ; Xiao-Ming FEI ; Jian-Yong LI
Journal of Experimental Hematology 2006;14(1):61-64
To explore the effects of proteasome inhibitor PS-341 on the cytokine expressions of mesenchymal stem cells (MSC) in patients with multiple myeloma (MM), MSCs of 11 patients were cultured in medium of RPMI 1640 containing 10% FBS. When cells grew to 5 x 10(5) - 1 x 10(6), cells were exposed to 50 nmol/L PS-341 for 4 hours, then harvested. The expressions of IL-6, IL-1beta and SCF were detected by RT-PCR. The results indicated that after treatment with PS-341 the expressions of IL-6, IL-1beta and SCF of MSCs decreased markedly, especially that of IL-1beta, compared with control (P < 0.05, P < 0.01, P < 0.05, respectively). There were obviously differences of IL-1beta expression between refractory/relapsed group and complete remission (CR) group and IL-1beta expression was inhibited more seriously in CR group, whereas there were no significant differences of IL-6 and SCF expression between two groups; IL-1beta expression of patients treated with PS-341 was not detected; there were not effects of IL-1beta expression on expressions of IL-6 and SCF. It is concluded that proteasome inhibitor PS-341 downregulated the expressions of IL-6, IL-1beta and SCF of MSCs in patients with MM.
Antineoplastic Agents
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pharmacology
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Bone Marrow Cells
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metabolism
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pathology
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Boronic Acids
;
pharmacology
;
Bortezomib
;
Cytokines
;
biosynthesis
;
Humans
;
Interleukin-1beta
;
biosynthesis
;
Interleukin-6
;
biosynthesis
;
Mesenchymal Stromal Cells
;
metabolism
;
Multiple Myeloma
;
metabolism
;
pathology
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Protease Inhibitors
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pharmacology
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Pyrazines
;
pharmacology
;
Stem Cell Factor
;
biosynthesis
2.Effects of baicalin on the expression of pro-MMP-1 and MMP-3 in human gingival fibroblasts and periodontal ligament cells.
Cheng-zhang LI ; Zheng-guo CAO ; Ru YANG ; Zhu-huan SHANG ; Li-jian JIN ; E F COBERT
Chinese Journal of Stomatology 2004;39(3):197-200
OBJECTIVETo investigate the influence of baicalin on the IL-1beta induced pro-MMP-1 in HGF and the effects of baicalin on MMP-3 expression in periodontal ligament cells (PDLCs).
METHODSThe amount of secreted pro-MMP-1 and MMP-3 expression was detected by ELISA and cell immunochemistry.
RESULTS(1) The amount of secreted pro-MMP-1 (3.333 +/- 0.123) microg/L increased significantly following 1 microg/L of IL-1beta, compared with control group (1.960 +/- 0.180) microg/L. Addition of baicalin to cell culture medium for 1 hour following IL-1beta decreased pro-MMP-1 secretion in a dose-dependent manner in the range of 10 approximately 1,000 microg/L. (2) 1 microg/L IL-1beta could significantly stimulate the synthesis and secretion of MMP-3 in PDLCs. (3) The baicalin could not interfere the synthesis of MMP-3, but could inhibit the release of MMP-3 from PDLCs.
CONCLUSIONSBaicalin could inhibit the secretion of pro-MMP-1 and MMP-3 expression in IL-1beta induced HGF and PDLCs, which suggests that baicalin may play an important role in preventing and treating periodontal disease.
Collagenases ; biosynthesis ; genetics ; Enzyme Precursors ; biosynthesis ; genetics ; Fibroblasts ; enzymology ; pathology ; Flavonoids ; pharmacology ; Gingiva ; enzymology ; pathology ; Humans ; Interleukin-1 ; pharmacology ; Interleukin-1beta ; Matrix Metalloproteinase 1 ; Metalloendopeptidases ; biosynthesis ; genetics ; Peptide Fragments ; pharmacology ; Periodontal Ligament ; enzymology ; pathology ; Periodontitis ; enzymology ; pathology ; Scutellaria ; chemistry
3.Effect of lipoxin A₄ on IL-1β production of monocytes and its possible mechanism in severe preeclampsia.
Jianfang WANG ; Yinping HUANG ; Yanjun HUANG ; Jie ZHOU ; Xiaoli LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(6):767-770
This study examined in vitro effect of lipoxin A(4) (LXA(4)) on interleukin-1β (IL-1β) production of monocytes and its possible mechanism in severe preeclampsia (PE). Peripheral venous blood was drawn from 15 patients with severe preeclampsia (PE group) and 20 normal pregnant women (control group) to prepare monocytes which were then treated with LXA(4) at different concentrations of 0, 10, 100 nmol/L respectively. IL-1β level in the supernatant of monocytes was detected by enzyme linked immunoassay. The [Ca(2+)](i) of monocytes was measured by laser scanning confocal microscopy. The results showed that the IL-1β level and the [Ca(2+)](i) of monocytes in the PE group were significantly higher than those in the control group. LXA(4) significantly decreased the generation of IL-1β in a dose-dependent manner in the PE group. After treatment with 100-nmol/L LXA(4), in the PE group, the [Ca(2+)](i) concentration of monocytes was significantly reduced. It was concluded that LXA(4) may inhibit the IL-1β production of monocytes from severe preeclampsia women by inhibiting extracellular calcium influx.
Adult
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Anti-Inflammatory Agents, Non-Steroidal
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pharmacology
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Calcium
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metabolism
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Cells, Cultured
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Female
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Humans
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Interleukin-1beta
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biosynthesis
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Lipoxins
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pharmacology
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Monocytes
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cytology
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metabolism
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Pre-Eclampsia
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blood
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physiopathology
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Pregnancy
4.Activated T cells promote differentiation of mesenchymal stem cells into osteoblasts.
Yuan-Lin LIU ; Xiao-Xia JIANG ; Yong-Feng SU ; Si-Wei HUO ; Heng ZHU ; Ying WU ; Xiu-Sen LI ; Ning MAO ; Yi ZHANG
Journal of Experimental Hematology 2009;17(4):974-976
This study was aimed to investigate the effect of activated T cell on the ability of MSC to differentiate into osteoblasts. The activated T cells with MSCs were co-culture for 14 days, then the osteoblast formation was tested by alkaline phosphatase staining. Furthermore, the supernatant of activated T cell was added in culture system of MSCs, the expression of molecules related with immune regulation of activated T cells was detected by RT-PCR, so as to determine what kinds of cytokine displayed the important function in MSC differentiation. The result showed that activated T cell could promote differentiation of MSC into osteoblasts, and IL-1beta played an important role in the effect of activated T cells on MSCs, while TNF-alpha, TGF-beta1 were not. It is concluded that the activated T cells promote the differentiation of MSCs to osteoblasts. The interactive influence between MSCs and immune cells can be mediated through cytokines.
Cell Differentiation
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Cells, Cultured
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Coculture Techniques
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Culture Media, Conditioned
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Humans
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Interleukin-1beta
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biosynthesis
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Mesenchymal Stromal Cells
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cytology
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Osteoblasts
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cytology
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T-Lymphocytes
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metabolism
5.Effects of interleukin-1beta on nitric oxide production and ATP synthesis in rat surplus hepatocytes after partial hepatectomy.
Wei TU ; Song HU ; Man ZHAO ; Wen-zhi QU ; Zuo-fu YU ; Wen-yu SUN
Acta Academiae Medicinae Sinicae 2007;29(5):631-637
OBJECTIVETo detect the nitric oxide (NO) production and energy metabolism of the interleukin (IL)-1beta-treated residual hepatocytes from rats after partial hepatectomy.
METHODSForty rats were equally divided into partial hepatectomies (PH) group and control group. In the control group the rats were otherwise matched and underwent sham surgeries. The residual hepatocytes were separated by the collagenase perfusion method. The hepatocytes were cultured with cytokines such as IL-1beta. The production of NO in the two groups were measured with Griess reagent method, the production of inducible nitric oxide synthase (iNOS) protein detected with Western blot, the content of the nucleotide in the hepatocytes detected with high-performance liquid chromatography, and the content of the ketone body in the hepatocytes of the two groups determined with the enzymatic method. Afterwards the ketone body ratio (acetoacetate/beta-hydroxy butyrate, KBR) was calculated.
RESULTSThe production of NO in the PH group was twice as much as that in the Sham group. IL-1beta decreased the content of ATP and the KBR in the hepatocytes of both groups, and the decrease magni tude in the PH group was significantly larger than that in the Sham group. After the injection of L-arginine, the production of NO in the hepatocytes in the PH group increased, and the level of ATP and KBR decreased. N(G)-methyl-L-arginine (L-NMMA), the inhibitor of NO synthase, inhibited the production of NO and reversed the decrease of ATP and KBR.
CONCLUSIONAfter partial hepatectomy, increased NO production in the hepatocytes after the treatment of interleukin-1beta may disturb the function of mitochondria by inhibiting the synthesis of ATP.
Adenosine Triphosphate ; biosynthesis ; Animals ; Arginine ; pharmacology ; Cells, Cultured ; Hepatectomy ; Hepatocytes ; metabolism ; Interleukin-1beta ; pharmacology ; Ketone Bodies ; biosynthesis ; Nitric Oxide ; antagonists & inhibitors ; biosynthesis ; Nitric Oxide Synthase ; antagonists & inhibitors ; Rats ; omega-N-Methylarginine ; pharmacology
6.Effects of Ginkgo biloba extract on expressions of IL-1beta, TNF-alpha, and IL-10 in U937 foam cells.
Ya-Bin JIAO ; Yao-Cheng RUI ; Peng-Yuan YANG ; Tie-Jun LI ; Yan QIU
Acta Pharmaceutica Sinica 2007;42(9):930-934
This study is to investigate the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in U937 foam cells and effects of Ginkgo biloba extract (GbE) on the cytokines. U937 cells were cultured with different concentrations of GbE (0.1, 1, and 10 microg x L(-1)), and stimulated by 100 mg x L(-1) oxidized low density lipoprotein (ox-LDL) for 24 h. The expressions of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) in culture solution were detected by enzyme-linked immunosorbant assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that incubated with 100 mg x L(-1) ox-LDL for 24 h, the U937 cells became foam cells, the protein or mRNA expressions of IL-1beta, TNF-alpha, IL-10, and its receptor IL-10R in U937 foam cells were higher markedly than those in normal U937 cells. When the cells were pretreated with GbE (0.1, 1, and 10 microg x L(-1)), the increases of IL-1beta and TNF-alpha in U937 foam cells were remarkably inhibited, but IL-10 expression increased greatly. Especially when cells were pretreated with 10 microg x L(-1) GbE, the protein and mRNA expressions of IL-1beta and TNF-alpha were markedly lower than those in U937 foam cells. The protein expression of IL-10 and mRNA expressions of IL-10 and its receptor IL-10R were markedly higher than those in U937 foam cells. GbE inhibited production of pro-inflammatory cytokines IL-1beta and TNF-alpha, but up-regulated the production of anti-inflammatory cytokine IL-10 and its receptor IL-10R in U937 foam cells, which might be related with its anti-atherosclerotic actions.
Drugs, Chinese Herbal
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isolation & purification
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pharmacology
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Foam Cells
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metabolism
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Ginkgo biloba
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chemistry
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Humans
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Interleukin-10
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biosynthesis
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genetics
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Interleukin-1beta
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biosynthesis
;
genetics
;
Lipoproteins, LDL
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Plants, Medicinal
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chemistry
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RNA, Messenger
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metabolism
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Receptors, Interleukin-10
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biosynthesis
;
genetics
;
Tumor Necrosis Factor-alpha
;
biosynthesis
;
genetics
;
U937 Cells
7.Effects of Armillariella tabescens polysaccharide IPS-B2 on activity of mouse peritoneal macrophages and transcription of related gene.
Xia LUO ; Meng-Yao YU ; Xiao-Yan XU ; Jin ZENG ; Nan JIANG ; Lin-Yong ZHENG
China Journal of Chinese Materia Medica 2008;33(11):1305-1308
OBJECTIVETo observe the effect of IPS-B2 on mouse peritoneal macrophages and the transcription of IL-1beta, IL-6, TNF-alpha and iNOS.
METHODELISA 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.
RESULTIPS-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.
CONCLUSIONIPS-B2 can enhance the ability of peritoneal macrophages to excrete bioactive substances and promote the transcription of bioactive substances to antitumor.
Agaricales ; chemistry ; Animals ; Gene Expression Regulation ; drug effects ; Interleukin-1beta ; biosynthesis ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; drug effects ; metabolism ; Mice ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Polymerase Chain Reaction ; Polysaccharides ; pharmacology ; RNA, Messenger ; biosynthesis ; Transcription, Genetic ; drug effects ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics
8.Endogenous danger signals trigger hepatic ischemia/reperfusion injury through toll-like receptor 4/nuclear factor-kappa B pathway.
Hui WANG ; Zhuo-ya LI ; He-shui WU ; Yang WANG ; Chun-fang JIANG ; Qi-chang ZHENG ; Jin-xiang ZHANG
Chinese Medical Journal 2007;120(6):509-514
BACKGROUNDRestoration of blood flow to the ischemic liver lobes may paradoxically exacerbate tissue injury, which is called hepatic ischemia/reperfusion injury (IRI). Toll-like receptor 4 (TLR4), expressed on several liver cell types, and the nuclear factor-kappa B (NF-kappaB) signaling pathway are crucial to mediating hepatic inflammatory response. Because IRI is essentially a kind of profound acute inflammatory reaction evoked by many kinds of danger signals, we investigated TLR4/NF-kappaB signaling pathway activation in a murine model of partial hepatic IRI.
METHODSWild-type mice (WT, C3H/HeN) or TLR4 mutant mice (C3H/HeJ) were subjected to 45 minutes of partial hepatic ischemia followed by 1 hour, 3 hours of reperfusion. Sham group accepted the same procedure without the obstruction of blood supply. At the end of reperfusion, the compromise of liver function and the histological change of liver sections were measured as the severity of liver injury. The level of endotoxin in the portal vein was measured by limulus assay. NF-kappaB activation was determined by electrophoretic mobility shift assay (EMSA). The levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in systemic blood after hepatic IRI were assessed by enzyme-linked immunosorbent assay (ELISA).
RESULTSThe compromise of liver function and the morphological injuries in mutant mice were relieved more markedly than those in WT mice after partial hepatic IRI. NF-kappaB activation in WT mice was stronger than that in TLR4 mutant mice, and both were stronger than those in the sham operated mice (P < 0.01). Endotoxin in each group was undetectable. The levels of TNF-alpha and IL-1beta in systemic blood were elevated in both strains, but lower in the sham operated group. These mediators were significantly decreased in TLR4 mutant mice compared with those in WT mice (P < 0.01).
CONCLUSIONSThe TLR4/NF-kappaB signaling pathway may mediate hepatic IRI triggered by endogenous danger signals. Inhibition of the TLR4/NF-kappaB pathway may be a potential therapeutic target for attenuating ischemia/reperfusion-induced tissue damage in some clinical settings.
Alanine Transaminase ; blood ; Animals ; Interleukin-1beta ; biosynthesis ; Liver ; blood supply ; Mice ; Mice, Inbred C3H ; NF-kappa B ; physiology ; Reperfusion Injury ; etiology ; Signal Transduction ; physiology ; Toll-Like Receptor 4 ; physiology ; Tumor Necrosis Factor-alpha ; biosynthesis
9.Role of cell-surface nucleolin in lipopolysaccharide-stimulated expression and secretion of TNF-alpha and IL-1beta.
Li FANG ; Kang-kai WANG ; Lei JIANG ; Bi-mei JIANG ; Xing WEI ; Lan SONG ; Gong-hua DENG ; Xian-zhong XIAO
Journal of Central South University(Medical Sciences) 2008;33(11):999-1004
OBJECTIVE:
To explore the role of cell-surface nucleolin in lipopolysaccharide (LPS)-stimulated expression and secretion of TNF-alpha and IL-1beta in human THP-1 monocytes.
METHODS:
Immuno-fluorescence assay and Western blot were used to identify the expression of nucleolin on the surface of THP-1 monocytes. Inactivation of nucleolin was induced by anti-nucleolin monoclonal antibody blockage, and the expression and secretion of TNF-alpha and IL-1beta were observed by using reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immuno-sorbent assay (ELISA)respectively in LPS-mediated human THP-1 monocyte inflammatory model.
RESULTS:
Immuno-fluorescence showed that nucleolin was localized on the cell surface of THP-1 monocytes as indicated by dotted red fluorescence. Western blot assay indicated that nucleolin existed in the cell membrane fractions. RT-PCR assay showed that the expressions of TNF-alpha and IL-1beta mRNA significantly increased at 2 h and 3 h after the treatment with 1000 microg/L LPS. After 1 h pretreatment with anti-nucleolin antibody, the levels of TNF-alpha and IL-1beta mRNA decreased compared with an anti-nucleolin antibody untreated group and an irrelevant IgG+LPS group (P<0.05). ELISA assay showed that the pretreatment with anti-nucleolin antibody inhibited significantly the secretion of LPS-induced levels of TNF-alpha and IL-1beta after 4, 12 and 24 h treatment with 1000 microg/L LPS.
CONCLUSION
Nucleolin expresses on the cell surface of THP-1 monocytes and involves in the LPS-mediated expression and secretion of TNF-alpha and IL-1beta.
Cell Line
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Cell Membrane
;
metabolism
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Humans
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Interleukin-1beta
;
biosynthesis
;
metabolism
;
Lipopolysaccharides
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pharmacology
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Monocytes
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cytology
;
metabolism
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Phosphoproteins
;
metabolism
;
physiology
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RNA-Binding Proteins
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metabolism
;
physiology
;
Tumor Necrosis Factor-alpha
;
biosynthesis
;
metabolism
10.Telmisartan reduced cerebral edema by inhibiting NLRP3 inflammasome in mice with cold brain injury.
Xin WEI ; Chen-Chen HU ; Ya-Li ZHANG ; Shang-Long YAO ; Wei-Ke MAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2016;36(4):576-583
The aim of this study was to investigate the possible beneficial role of telmisartan in cerebral edema after traumatic brain injury (TBI) and the potential mechanisms related to the nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) pyrin domain-containing 3 (NLRP3) inflammasome activation. TBI model was established by cold-induced brain injury. Male C57BL/6 mice were randomly assigned into 3, 6, 12, 24, 48 and 72 h survival groups to investigate cerebral edema development with time and received 0, 5, 10, 20 and 40 mg/kg telmisartan by oral gavage, 1 h prior to TBI to determine the efficient anti-edemic dose. The therapeutic window was identified by post-treating 30 min, 1 h, 2 h and 4 h after TBI. Blood-brain barrier (BBB) integrity, the neurological function and histological injury were assessed, at the same time, the mRNA and protein expression levels of NLRP3 inflammasome, IL-1β and IL-18 concentrations in peri-contused brain tissue were measured 24 h post TBI. The results showed that the traumatic cerebral edema occurred from 6 h, reached the peak at 24 h and recovered to the baseline 72 h after TBI. A single oral dose of 5, 10 and 20 mg/kg telmisartan could reduce cerebral edema. Post-treatment up to 2 h effectively limited the edema development. Furthermore, prophylactic administration of telmisartan markedly inhibited BBB impairment, NLRP3, apoptotic speck-containing protein (ASC) and Caspase-1 activation, as well as IL-1β and IL-18 maturation, subsequently improved the neurological outcomes. In conclusion, telmisartan can reduce traumatic cerebral edema by inhibiting the NLRP3 inflammasome-regulated IL-1β and IL-18 accumulation.
Animals
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Benzimidazoles
;
administration & dosage
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Benzoates
;
administration & dosage
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Blood-Brain Barrier
;
drug effects
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Brain Edema
;
drug therapy
;
genetics
;
pathology
;
Brain Injuries, Traumatic
;
drug therapy
;
genetics
;
pathology
;
Caspase 1
;
biosynthesis
;
Gene Expression Regulation
;
drug effects
;
Humans
;
Inflammasomes
;
adverse effects
;
genetics
;
Interleukin-18
;
biosynthesis
;
Interleukin-1beta
;
biosynthesis
;
Male
;
Mice
;
NLR Family, Pyrin Domain-Containing 3 Protein
;
biosynthesis
;
genetics
;
Signal Transduction
;
drug effects