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1.
Hum Exp Toxicol ; 40(12): 2048-2062, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34053323

ABSTRACT

PURPOSE: The mechanism of cytotoxicity of silibinin on two human hepatocellular carcinoma (HCC) cell lines, HepG2 (p53 wild-type) and Hep3B cells (p53 null), is examined in relation with the induction of autophagy and phosphorylation of AMP-activated protein kinase (p-AMPK). MATERIALS AND METHODS: Levels of apoptosis in relation to the levels of autophagy and those of glycolysis-related proteins, glucose transporter 1/4 (Glut1/4) and hexokinase-II (HK2), in HepG2 and Hep3B cells were examined. RESULTS: Silibinin-induced apoptosis was incomplete for HCC cell death in that up-regulated autophagy and/or reduced level of glycolysis, which are induced by silibinin treatment, antagonized silibinin-induced apoptosis. Inhibition of autophagy with 3-methyl adenine (3MA) or blocking of AMP-activated protein kinase (AMPK) activation with Compound C (CC) enhanced silibinin-induced apoptosis. The results confirm that AMPK involved in autophagy as well as in glycolysis remaining with silibinin is responsible for attenuation of silibinin-induced apoptosis. Blocking of AMPK or autophagy contributes to the enhancement of silibinin's cytotoxicity to HepG2 and Hep3B cells. CONCLUSION: This study shows that incomplete apoptosis of HCC by silibinin treatment becomes complete by repression of autophagy and/or glycolysis.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Carcinoma, Hepatocellular/drug therapy , Liver Neoplasms/drug therapy , Silymarin/pharmacology , AMP-Activated Protein Kinases/metabolism , Apoptosis/drug effects , Autophagy/drug effects , Carcinoma, Hepatocellular/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Glycolysis/drug effects , Humans , Liver Neoplasms/metabolism , Phosphorylation/drug effects , Poly(ADP-ribose) Polymerases/metabolism
2.
J Pharmacol Exp Ther ; 331(1): 319-26, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19638571

ABSTRACT

In Alzheimer's disease (AD), the deposition of amyloid peptides is invariably associated with oxidative stress and inflammatory responses. Silibinin (silybin), a flavonoid derived from the herb milk thistle, has potent anti-inflammatory and antioxidant activities. However, it remains unclear whether silibinin improves amyloid beta (Abeta) peptide-induced neurotoxicity. In this study, we examined the effect of silibinin on the fear-conditioning memory deficits, inflammatory response, and oxidative stress induced by the intracerebroventricular injection of Abeta peptide(25-35) (Abeta(25-35)) in mice. Mice were treated with silibinin (2, 20, and 200 mg/kg p.o., once a day for 8 days) from the day of the Abeta(25-35) injection (day 0). Memory function was evaluated in cued and contextual fear-conditioning tests (day 6). Nitrotyrosine levels in the hippocampus and amygdala were examined (day 8). The mRNA expression of inducible nitric-oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-alpha) in the hippocampus and amygdala was measured 2 h after the Abeta(25-35) injection. We found that silibinin significantly attenuated memory deficits caused by Abeta(25-35) in the cued and contextual fear-conditioning test. Silibinin significantly inhibited the increase in nitrotyrosine levels in the hippocampus and amygdala induced by Abeta(25-35). Nitrotyrosine levels in these regions were negatively correlated with memory performance. Moreover, real-time RT-PCR revealed that silibinin inhibited the overexpression of iNOS and TNF-alpha mRNA in the hippocampus and amygdala induced by Abeta(25-35). These findings suggest that silibinin (i) attenuates memory impairment through amelioration of oxidative stress and inflammatory response induced by Abeta(25-35) and (ii) may be a potential candidate for an AD medication.


Subject(s)
Amyloid beta-Peptides/toxicity , Memory Disorders/metabolism , Memory Disorders/prevention & control , Nitric Oxide Synthase Type II/biosynthesis , Peptide Fragments/toxicity , Tumor Necrosis Factor-alpha/biosynthesis , Animals , Antioxidants/pharmacology , Antioxidants/therapeutic use , Drug Synergism , Inflammation Mediators/therapeutic use , Male , Memory Disorders/chemically induced , Memory Disorders/enzymology , Mice , Mice, Inbred ICR , Nitric Oxide Synthase Type II/antagonists & inhibitors , Nitric Oxide Synthase Type II/genetics , RNA, Messenger/biosynthesis , Silybin , Silymarin/pharmacology , Silymarin/therapeutic use , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Tumor Necrosis Factor-alpha/genetics , Tyrosine/analogs & derivatives , Tyrosine/biosynthesis , Up-Regulation/drug effects , Up-Regulation/genetics
3.
Br J Pharmacol ; 157(7): 1270-7, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19552690

ABSTRACT

BACKGROUND AND PURPOSE: Accumulated evidence suggests that oxidative stress is involved in amyloid beta (Abeta)-induced cognitive dysfunction. Silibinin (silybin), a flavonoid derived from the herb milk thistle (Silybum marianum), has been shown to have antioxidative properties; however, it remains unclear whether silibinin improves Abeta-induced neurotoxicity. In the present study, we examined the effect of silibinin on the memory impairment and accumulation of oxidative stress induced by Abeta(25-35) in mice. EXPERIMENTAL APPROACH: Aggregated Abeta(25-35) (3 nmol) was intracerebroventricularly administered to mice. Treatment with silibinin (2, 20 and 200 mg.kg(-1), once a day, p.o.) was started immediately after the injection of Abeta(25-35). Locomotor activity was evaluated 6 days after the Abeta(25-35) treatment, and cognitive function was evaluated in a Y-maze and novel object recognition tests 6-11 days after the Abeta(25-35) treatment. The levels of lipid peroxidation (malondialdehyde) and antioxidant (glutathione) in the hippocampus were measured 7 days after the Abeta(25-35) injection. KEY RESULTS: Silibinin prevented the memory impairment induced by Abeta(25-35) in the Y-maze and novel object recognition tests. Repeated treatment with silibinin attenuated the Abeta(25-35)-induced accumulation of malondialdehyde and depletion of glutathione in the hippocampus. CONCLUSIONS AND IMPLICATIONS: Silibinin prevents memory impairment and oxidative damage induced by Abeta(25-35) and may be a potential therapeutic agent for Alzheimer's disease.


Subject(s)
Amyloid beta-Peptides/physiology , Memory Disorders/drug therapy , Neuroprotective Agents/pharmacology , Oxidative Stress/drug effects , Peptide Fragments/physiology , Alzheimer Disease/drug therapy , Amyloid beta-Peptides/toxicity , Animals , Exploratory Behavior/drug effects , Glutathione/metabolism , Hippocampus/metabolism , Lipid Peroxidation/drug effects , Male , Malondialdehyde/metabolism , Maze Learning/drug effects , Memory Disorders/chemically induced , Mice , Mice, Inbred ICR , Motor Activity/drug effects , Neuroprotective Agents/therapeutic use , Peptide Fragments/toxicity , Recognition, Psychology/drug effects , Silybin , Silymarin/pharmacology , Silymarin/therapeutic use
4.
J Asian Nat Prod Res ; 9(3-5): 245-52, 2007.
Article in English | MEDLINE | ID: mdl-17566917

ABSTRACT

Silymarin, derived from the milk thistle plant, Silybum marianum, has been traditionally used in the treatment of liver disease. Our previous study demonstrated that silymarin has an anti-apoptotic effect against UV irradiation. In this study, SIRT1, a human deacetylase that was reported to promote cell survival, was activated by silymarin (5 x 10(- 4) mol/L) in UV-irradiated human malignant melanoma, A375-S2 cells, followed by down-regulated expression of Bax and decreased release of cytochrome c. Cleavage of procaspase-3 and digestion of its substrates, the inhibitor of caspase-activated DNase (ICAD) and poly(ADP-ribose) polymerase (PARP), were also reduced. Consistent with its protective effect on UV-induced apoptosis, silymarin (5 x 10(- 4) mol/L) also increased G(2)/M phase arrest, possibly providing a prolonged time for efficient DNA repair. Consequently, that silymarin protected A375-S2 cell against UV-induced apoptosis was partially through SIRT1 pathway and modulation of the cell cycle distribution.


Subject(s)
Apoptosis/radiation effects , Cell Cycle/drug effects , Cytoprotection , Silymarin/pharmacology , Sirtuins/physiology , Cell Line, Tumor , Humans , Sirtuin 1 , Ultraviolet Rays
5.
J Asian Nat Prod Res ; 8(4): 335-43, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16864444

ABSTRACT

Dracorhodin perchlorate, an anthocyanin red pigment, induces human premyelocytic leukemia HL-60 cell death through apoptotic pathway. Caspase -1, -3, -8, -9, and -10 inhibitors partially reversed the cell death induced by dracorhodin perchlorate. Caspase-3 and -8 were activated followed to the degradation of caspase-3 substrates, inhibitor of caspase-activated DNase (ICAD) and poly-(ADP-ribose) polymerase (PARP). Dracorhodin perchlorate up-regulated the expression ratio of mitochondrial proteins, Bax/Bcl-XL. The cell death was accompanied with phosphorylation of ERK, JNK and p38 MAPK and partially reduced by MEK inhibitor (PD98059), JNK MAPK inhibitor (SP600125) and p38 MAPK inhibitor (SB 203580). Taken together, dracorhodin perchlorate-induced apoptosis in HL-60 cells via up-regulation of Bax, activation of caspases and ERK/p38/JNK MAPKs.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Benzopyrans/pharmacology , Antineoplastic Agents, Phytogenic/chemistry , Benzopyrans/chemistry , Caspases/metabolism , Dose-Response Relationship, Drug , Enzyme Activation , HL-60 Cells , Humans , MAP Kinase Kinase 4/metabolism , Mitogen-Activated Protein Kinase Kinases/metabolism , Molecular Structure , bcl-2-Associated X Protein/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism
6.
Trans R Soc Trop Med Hyg ; 99(8): 625-30, 2005 Aug.
Article in English | MEDLINE | ID: mdl-15927218

ABSTRACT

Neurocysticercosis (NCC) is one of the major causes of neurological disease in China. ELISA and immunoblotting using glycoproteins purified by preparative isoelectric focusing were used to detect human cysticercosis in Tongliao area, Inner Mongolia, China in 1998. Approximately 89% (39 of 44 inpatients and outpatients with suspected NCC at Tongliao City Hospital) were residents of Inner Mongolia. About 53% were male and 77% were of working age (18-59 years), and 32% were farmers. Immunoblotting and ELISA showed a high correlation. Of the 44 patients, 31 positive by cerebral computed tomography (CT) scan were confirmed serologically to have cysticercosis. In the ELISA, patients with no lesions by CT scan had lower OD values, similar to those of normal serum. These findings confirm that both ELISA and immunoblotting assays are sufficiently sensitive to detect asymptomatic or symptomatic cysticercosis patients.


Subject(s)
Cysticercosis/diagnosis , Cysticercosis/epidemiology , Neurocysticercosis/diagnosis , Taenia solium/immunology , Adolescent , Adult , Age Distribution , Animals , Antibodies, Helminth/blood , China/epidemiology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoblotting , Male , Middle Aged , Neurocysticercosis/complications , Neurocysticercosis/epidemiology , Sensitivity and Specificity , Sex Distribution
7.
Pharmazie ; 59(4): 294-6, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15125576

ABSTRACT

A new steroidal saponin, dioscoreside E (1), and a known compound, protodioscin (2), were isolated from an ethanol extract of the rhizomes of Dioscorea panthaica. The structure of 1 was established as 3-O-[bis-alpha-L-rhamnopyranosyl-(1 --> 2 and 1 --> 4)-beta-D-glucopyranosyl]-26-O-beta-D-glucopyranosyl-20(R)-methoxy-25(R)-furosta-5,22(23)-diene-3beta,26-diol, on the basis of spectral and chemical evidence. Compounds 1 and 2 showed cytotoxic activity against a panel of tumor cell lines.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Dioscorea/chemistry , Saponins/chemistry , Saponins/isolation & purification , Saponins/pharmacology , Steroids/chemistry , Steroids/isolation & purification , Steroids/pharmacology , Animals , Cell Line, Tumor , Drug Screening Assays, Antitumor , Humans , Hydrolysis , Magnetic Resonance Spectroscopy , Mice , Models, Molecular , Molecular Conformation , Spectrometry, Mass, Fast Atom Bombardment
8.
Planta Med ; 67(9): 853-7, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11745024

ABSTRACT

Two new steroidal saponins, dioscoresides C (1) and D (2), along with a new natural product, pregnadienolone 3-O-beta-gracillimatriose (3), and two known compounds, pregnadienolone 3-O-beta-chacotrioside (4) and pseudoprotodioscin (5), were isolated from the rhizomes of Dioscorea panthaica Prain et Burkill. On the basis of extensive NMR studies and chemical evidence, dioscoresides C and D were determined to be 26-O-beta-D-glucopyranosyl-3 beta,26-dihydroxy-23(S)-methoxy-25(R)-furosta-5,20(22)-dien-3-O-alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-rhamnopyranosyl-(1-->4)]-beta-D-glucopyranoside and 26-O-beta-D-glucopyranosyl-3 beta,26-dihydroxy-20,22-seco-25(R)-furosta-5-en-20,22-dine-3-O-alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-rhamnopyranosyl-(1--> 4)]-beta-D-glucopyranoside. These compounds showed mild cytotoxicity against the cancer cell lines, A375, L929, and HeLa, in a dose-dependent manner.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Dioscorea , Rhizome/chemistry , Saponins/pharmacology , Steroids , Animals , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Cell Division/drug effects , Cell Line , Drugs, Chinese Herbal , HeLa Cells , Humans , Magnetic Resonance Spectroscopy , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Saponins/chemistry , Saponins/isolation & purification , Tumor Cells, Cultured
9.
J Clin Microbiol ; 38(12): 4439-44, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11101577

ABSTRACT

Neurocysticercosis (NCC) caused by infection with the larvae of Taenia solium is an important cause of neurological disease worldwide. In order to establish an enzyme-linked immunosorbent assay (ELISA) for this infection using recombinant proteins, we carried out molecular cloning and identified four candidates as diagnostic antigens (designated Ag1, Ag1V1, Ag2, and Ag2V1). Except for Ag2V1, these clones could encode a 7-kDa polypeptide, and Ag2V1 could encode a 10-kDa polypeptide. All of the clones were very similar. Except for Ag2V1, recombinant proteins were successfully expressed using an Escherichia coli expression system. Immunoblot analysis of NCC patient sera detected recombinant proteins, but because reactivity to recombinant Ag1 was too weak, Ag1 was not suitable as an immunodiagnostic antigen. So, Ag1V1 and Ag2 were chosen as ELISA antigens, and the Ag1V1/Ag2 chimeric protein was expressed. Of 49 serum samples from NCC patients confirmed to be seropositive by immunoblot analysis, 44 (89.7%) were positive by ELISA. No assays of serum samples from patients with other parasitic infections recognized the Ag1V1/Ag2 chimeric protein. The Ag1V1/Ag2 chimeric protein obtained in this study had a high value for differential immunodiagnosis.


Subject(s)
Antigens, Helminth/genetics , Neurocysticercosis/diagnosis , Recombinant Fusion Proteins/immunology , Taenia/immunology , Amino Acid Sequence , Animals , Antigens, Helminth/immunology , Blotting, Southern , Cloning, Molecular , Enzyme-Linked Immunosorbent Assay , Molecular Sequence Data , Molecular Weight
10.
Immunopharmacology ; 49(3): 307-16, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10996028

ABSTRACT

The effect of T-cell-independent B cell mitogen, a pectic polysaccharide, bupleuran 2IIc, from a medicinal herb, the roots of Bupleurum falcatum L. on interleukin 6 (IL-6) production of murine B cells and B cell lines was investigated in order to clarify the mechanism of enhanced immunoglobulin (Ig) secretion from B cells. Bupleuran 2IIc enhanced IgM secretion from highly purified murine normal B cells. When normal B cells from murine spleen were cultured with bupleuran 2IIc in the presence of anti-IL-6 neutralizing antibody, the enhanced IgM secretion by bupleuran 2IIc was reduced. When B cells were stimulated with bupleuran 2IIc, their IL-6 secretion and the transcription of IL-6 mRNA were enhanced. The enhanced IL-6 secretion by bupleuran 2IIc was also observed in B cell line, Y16 cell. When bupleuran 2IIc was digested with endo-alpha-(1-->4)-D-polygalacturonase, the resulting enzyme resistant carbohydrate portion, "ramified" region (PG-1) composed of rhamnogalacturonan core containing neutral sugar side chains showed potent IL-6 secretion-enhancing activity. These results suggest that the "ramified" region of bupleuran 2IIc stimulates the secretion of IL-6 as the active site, and the resulting IL-6 may partially contribute the enhancement of IgM secretion as an autocrine and/or paracrine mechanism.


Subject(s)
Adjuvants, Immunologic/pharmacology , B-Lymphocytes/drug effects , Drugs, Chinese Herbal/pharmacology , Interleukin-6/biosynthesis , Pectins/pharmacology , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Adjuvants, Immunologic/chemistry , Animals , B-Lymphocytes/metabolism , Bupleurum , Cell Line , Drugs, Chinese Herbal/chemistry , Female , Immune Sera/pharmacology , Immunoglobulin M/biosynthesis , Interleukin-6/immunology , Interleukin-6/metabolism , Mice , Mice, Inbred C3H , Pectins/chemistry , Plant Extracts/chemistry , Plant Roots/chemistry , Structure-Activity Relationship , Tumor Cells, Cultured
11.
Acta Pharmacol Sin ; 21(12): 1179-85, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11603297

ABSTRACT

AIM: To compare the effects of water-soluble polysaccharides, FI0-b, and its formic acid-modified derivative, FI0-b-H, on production of human proinflammatory cytokines. METHODS: The polysaccharides were modified by formic acid. Cytokine production was quantitated by radioimmunoassay. mRNA for cytokines was measured by semi-quantitative RT-PCR. RESULTS: FI0-b and FI0-b-H 4, 40, and 400 mg/L significantly downregulated interleukin-1 alpha (IL-1 alpha) production by THP-1 cells induced by lypopolysaccharide (LPS) 1 or 10 mg/L and phorbol myristate acetate (PMA) 200 nmol/L. At lower stimulation with LPS 10 mg/L and PMA 200 nmol/L, both polysaccharides significantly upregulated tumor necrosis factor alpha (TNF alpha) production by THP-1 cells. However, at higher stimulation with LPS 100 mg/L and PMA 200 nmol/L, they downregulated TNF alpha production. FI0-b-H downregulated interleukin-8 (IL-8) production by THP-1 cells at a lower-dose of LPS 1 mg/L and PMA 200 nmol/L, but upregulated IL-8 production at a higher-dose of LPS 10 mg/L and PMA 200 nmol/L. Production of cytokines (IL-1 alpha and TNF alpha) was transcriptionally or post-transcriptionally regulated by FI0-b and FI0-b-H. CONCLUSION: The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different stimulatory conditions in a dose-dependent manner. Compared with FI0-b, FI0-b-H has more marked effects on human proinflammatory cytokine production.


Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Interleukin-1/biosynthesis , Mycelium/chemistry , Polysaccharides/pharmacology , Tumor Necrosis Factor-alpha/biosynthesis , Adult , Cell Separation , Humans , Interleukin-1/genetics , Interleukin-6/biosynthesis , Interleukin-8/biosynthesis , Leukemia, Myeloid/metabolism , Leukemia, Myeloid/pathology , Leukocytes, Mononuclear/metabolism , Polysaccharides/administration & dosage , Polysaccharides/isolation & purification , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Reishi , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/genetics
12.
Acta Pharmacol Sin ; 21(12): 1186-92, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11603298

ABSTRACT

AIM: To study the effects of water-soluble polysaccharides. FI0-c, and its sulfated derivative, FI0-c-S, on production of human proinflammatory cytokines, interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor alpha (TNF alpha). METHODS: The herbal polysaccharides were modified by chlorosulfornic acid in dimethyl sulfoxide (Me2SO). Cytokine production was measured by radioimmunoassay, mRNA for the cytokines was measured by semi-quantitative RT-PCR. RESULTS: FI0-c 4 mg/L itself induced IL-1 alpha production by THP-1 cells without stimulants, such as lipopolysaccharides (LPS) and phorbol myristate acetate (PMA). On the other hand, FI0-c and FI0-c-S inhibited the IL-1 alpha production by THP-1 cells with these stimulants. FI0-c and FI0-c-S significantly upregulated TNF alpha production by THP-1 cells without stimulants or at a low dose of LPS 10 mg/L and PMA 200 nmol/L, whereas these polysaccharides markedly downregulated the TNF alpha production by a high dose of LPS 100 mg/L and PMA. Human peripheral blood mononuclear cells (PBMC) responded to FI0-c and FI0-c-S in IL-1 alpha and TNF alpha production in a fashion similar to THP-1 cell responses. FI0-c 4 mg/L downregulated high-dose LPS- and PMA-induced IL-1 alpha or TNF alpha mRNA and their protein production by THP-1 cells. CONCLUSION: The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different cell stimulatory conditions. Chemical modification of this polysaccharide changed the intensity of regulatory effect on cytokine production.


Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Interleukin-1/biosynthesis , Mycelium/chemistry , Polysaccharides/pharmacology , Tumor Necrosis Factor-alpha/biosynthesis , Adult , Cell Separation , Humans , Interleukin-1/genetics , Leukemia, Myeloid/metabolism , Leukemia, Myeloid/pathology , Leukocytes, Mononuclear/metabolism , Polysaccharides/administration & dosage , Polysaccharides/isolation & purification , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Reishi , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/genetics
13.
Acta Pharmacol Sin ; 21(9): 792-6, 2000 Sep.
Article in English | MEDLINE | ID: mdl-11501159

ABSTRACT

AIM: To compare the effect of ginsenoside Rg1 and its metabolite Rh1 on proinflammatory cytokines and their mRNA expression by THP-1 cells. METHODS: Human peripheral blood mononuclear cells (PBMC) were incubated with Rg1 and Rh1 at concentrations of 0.1, 1, 10, and 100 mg/L, and the cell proliferation was measured 24 h after incubation. Radioimmunoassay (RIA) was used to detect the production of proinflammatory cytokines, TNF alpha, IL-1 alpha, and IL-8. TNF alpha mRNA level was detected by reverse transcription polymerase chain reaction (RT-PCR) after administration of Rg1 and Rh1. RESULTS: Rg1 and Rh1 (at concentration of 0.1, 1, 10, 100 mg/L) had no effect on PBMC proliferation. Rh1 1 mg/L could upregulate the productions of TNF (and IL-8 induced by lipopolysaccharides (LPS) 10 mg/L plus phorbol myristate acetate (PMA) 200 nmol/L, however, Rg1 showed an inhibitory effect on TNF alpha production induced by LPS 100 mg/L. Rg1 1 mg/L and Rh1 100 mg/L enhanced the production of IL-1 alpha level in THP-1 cells in the presence of LPS 10 mg/L. RT-PCR revealed that Rh1 stimulated TNF alpha mRNA expression in suitable stimulatory conditions. CONCLUSION: Rg1 and Rh1 have different effects on the production of cytokines produced THP-1 cells stimulated by LPS and PMA.


Subject(s)
Saponins/metabolism , Saponins/pharmacology , Tumor Necrosis Factor-alpha/biosynthesis , Ginsenosides , Humans , Interleukin-1/biosynthesis , Interleukin-8/biosynthesis , Leukemia, Myelomonocytic, Acute/pathology , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Tumor Cells, Cultured/metabolism , Tumor Necrosis Factor-alpha/genetics
14.
J Helminthol ; 73(4): 363-5, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10654408

ABSTRACT

The establishment of reliable serological methods for cysticercosis in pigs is important for the surveillance, control and prevention of taeniosis/cysticercosis in humans as well as in pigs to prevent economic loss. Both ELISA and immunoblot using glycoproteins (GPs) purified by a single step of preparative iso-electric focusing, which are highly useful for human cysticercosis, have been applied for a serological study in pigs naturally infected with Taenia solium. All sera from pigs showed similar responses to those in human cysticercosis. Therefore, it is expected that both ELISA and immunoblots using GPs would be useful in differentiating infected pigs from uninfected ones.


Subject(s)
Cysticercosis/veterinary , Swine Diseases/diagnosis , Animals , Cysticercosis/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/veterinary , Glycoproteins/immunology , Humans , Immunoblotting/methods , Immunoblotting/veterinary , Swine
15.
Z Naturforsch C J Biosci ; 53(5-6): 352-8, 1998.
Article in English | MEDLINE | ID: mdl-9679326

ABSTRACT

Tissue type (type 2) transglutaminase (TGase, EC 2.3.2.13) has been implicated in various cellular processes including cell death. In order to better understand the role of this enzyme in cell death, human melanocytic A375-S2 cells were treated with sphingosine, a cell-signaling mediator. During the rapid onset of cytotoxicity caused by this lipidic agent, tissue TGase was translocated from the cytoplasm to the cell nuclei. This observation was further remarked in relevance to its previously undescribed activity for DNA degradation. The DNA hydrolytic activity associated with tissue TGase was dependent on Mg2+ in contrast to the Ca2+ requirement for the classical cross-linking activity of TGase, and was inhibited by Zn2+. Based on the results shown here, we propose a novel aspect of tissue TGase in cell death.


Subject(s)
Cell Nucleus/enzymology , Sphingosine/toxicity , Transglutaminases/metabolism , Cell Death/drug effects , Cell Survival/drug effects , Cytoplasm/enzymology , DNA, Neoplasm/metabolism , Humans , Hydrolysis , Melanoma , Substrate Specificity , Tumor Cells, Cultured
16.
J Interferon Cytokine Res ; 16(12): 1079-88, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8974012

ABSTRACT

On the basis of their relative hydropathy and alpha-helical structure, we prepared antibodies to four synthetic peptides with amino acid sequences homolgous to four hydrophilic, extracellular regions of the murine 80 kDa type I interleukin-1 receptor (IL-1RI). Antibodies to each of the four peptides recognized their specific immunogen. Human [125I]-IL-1 alpha or -beta was crosslinked to murine EL4 and D10S cells. Antiserum to peptide 150-166 precipitated the IL-1/IL-1R complex, whereas antibodies to peptide 66-84, 190-200, or 266-285 did not. Antibody to peptide 150-166 did not precipitate the type II IL-1R. Anti-IL-1RI150-166 blocked 71% of the binding of radiolabeled human IL-1 beta to EL4 cells and 50% of the binding to D10S cells. Using affinity-purified anti-IL-1RI150-166, we compared the ability of this antibody to inhibit the binding of murine or human IL-1 alpha to that of murine or human IL-1 beta. At a concentration of 20 ng/ml, affinity-purified anti-IL-1RI150-166 blocked 50% binding of murine IL-1 beta. At 1 microgram/ml, 90% blockage was observed. In contrast, no significant blockade of IL-1 alpha binding was observed at concentrations as high as 3 micrograms/ml of anti-IL-1RI150-166. The selective blockade of IL-1 beta forms was not due to differences in the affinities of these ligands for receptors on these cells. The antibody also blocked the binding of human IL-1 beta but not human IL-1 alpha to EL4 cells. The biologic activity of murine IL-1 beta but not IL-1 alpha on EL4 cells was also inhibited by this antibody. These data suggest (1) that antibody to a specific epitope on the extracellular domain interferes with the binding of IL-1 beta but not IL-1 alpha, (2) the differential inhibition of binding of IL-1 beta but not IL-1 alpha by anti-IL-1RI150-166 also blocks biologic activity, and (3) IL-1 alpha and IL-1 beta may transduce different signals by binding to separate loci on the IL-1RI.


Subject(s)
Antigen-Antibody Reactions , Interleukin-1/metabolism , Peptide Fragments/immunology , Protein Structure, Secondary , Receptors, Interleukin-1/immunology , Amino Acid Sequence , Animals , Binding Sites , Cell Line , Humans , Immunoglobulins/chemistry , Mice , Molecular Sequence Data , Molecular Weight , Precipitin Tests , Receptors, Interleukin-1/chemistry , Receptors, Interleukin-1/metabolism , Solubility , Tumor Cells, Cultured , Water/chemistry
18.
Nihon Ika Daigaku Zasshi ; 62(4): 386-90, 1995 Aug.
Article in English | MEDLINE | ID: mdl-7559927

ABSTRACT

It has been reported that the HMG-CoA reductase inhibitor simvastatin does not always effectively lower plasma LDL. This drug acts to monocytes/macrophages directly and inhibits cholesterol ester accumulation in these cells. However cytokine production in macrophages when simvastatin was administrated has not been described. In this study, we examined whether simvastatin affects cytokine production in human monocyte-derived macrophages. Simvastatin at doses ranging from 10(-9) to 10(-5) M did not affect the synthesis of proinflammatory cytokines (IL-1 beta, IL-6, IL-8) from human peripheral mononuclear cells. In addition, any changes in cytokine-induced cytokine production (IL-1-induced IL-8 synthesis) were not detected after the addition of simvastatin. The present results suggest that simvastatin suppresses foam cell formation in monocyte/macrophage, without affecting the immunological or inflammatory functions of these cells.


Subject(s)
Cytokines/biosynthesis , Enzyme Inhibitors/pharmacology , Hydroxymethylglutaryl-CoA Reductase Inhibitors , Lovastatin/analogs & derivatives , Macrophages/metabolism , Monocytes/metabolism , Cells, Cultured , Cholesterol Esters/metabolism , Humans , Lovastatin/pharmacology , Male , Simvastatin
19.
Lymphokine Cytokine Res ; 11(4): 197-205, 1992 Aug.
Article in English | MEDLINE | ID: mdl-1420598

ABSTRACT

The two types of cell surface receptors for interleukin-1 (IL-1) are glycoproteins that contain N-glycosidic chains on their extracellular portions. To determine the role of glycosylation of the IL-1 receptor type I (IL-1RtI) in the binding and function of IL-1, we used four plant lectins and glycosidase treatment on two different T-cell lines (EL4-6.1 and D10S) and expressing high number of binding sites for IL-1. The lectins wheat germ agglutinin, phytohemagglutinin, and concanavalin A inhibited in a dose-response manner the IL-1-induced proliferation of D10S cells. Binding of IL-1 was blocked and radioactive IL-1 was displaced from these cells by these lectins. Specific sugars (GlcNAc, NeuAc, Gal-GlcNAc-Man, Man, or alpha-MeMan) did not themselves affect IL-1 binding but reversed the blocking effects of the lectins. The two cell lines differed in their responses to the lectin-mediated inhibition of IL-1 binding. Digestion by N-glycosidase significantly decreased the capacity of cells to bind IL-1, and reduced by approximately 20,000 D the M(r) of the IL-1RtI. Neuraminidase and O-glycanase treatment did not alter the binding of IL-1 to D10S or EL4-6.1 cells. This study demonstrates that glycosylation of the extracellular domain of the IL-1RtI is due to N-linked carbohydrates, that the degree of glycosylation can vary in cells of different lineage, and that this N-linked glycosylation appears to be essential for optimal binding and activity of IL-1 to its type I receptor.


Subject(s)
Interleukin-1/metabolism , Receptors, Interleukin-1/metabolism , Animals , Antibodies, Monoclonal , Binding, Competitive , Carbohydrate Conformation , Cell Line , Glycosylation , Interleukin-1/antagonists & inhibitors , Lectins , Mice , Protein Binding , T-Lymphocytes/metabolism
20.
J Infect Dis ; 162(1): 215-23, 1990 Jul.
Article in English | MEDLINE | ID: mdl-2113076

ABSTRACT

Recombinant human interleukin-1 (IL-1), injected into rabbits, induces the synthesis of endogenous IL-1. Also, IL-1 induces its own gene expression and synthesis in human peripheral blood mononuclear cells (PBMC). In this study, tumor necrosis factor-alpha (TNF-alpha) production by PBMC of 40 individuals stimulated with IL-1 alpha or IL-1 beta was determined by specific radioimmunoassay (RIA). After 3 h of PBMC incubation with IL-1, TNF-alpha mRNA was detected. IL-1 alpha stimulated both IL-1 beta and TNF-alpha, but there was no correlation in the amount of TNF-alpha or IL-1 beta synthesized in the PBMC of 29 individuals. IL-1-stimulated adherent cells produced approximately 50% more TNF-alpha than did unfractionated PBMC. Coincubation with interferon-gamma (IFN-gamma) did not change the amount of IL-1-induced TNF-alpha, whereas in the same culture IFN-gamma inhibited (greater than 70%) IL-1-induced IL-1 production. Endogenous pyrogen and TNF-like activity were detected in the sera of rabbits 3.5 h after injection of either IL-1 alpha or -1 beta. These studies demonstrate that IL-1 induced TNF-alpha production in vivo and in vitro.


Subject(s)
Interferon-gamma/pharmacology , Interleukin-1/pharmacology , Leukocytes, Mononuclear/immunology , Tumor Necrosis Factor-alpha/biosynthesis , Animals , Blotting, Northern , Cell Line , Cells, Cultured , Cytotoxicity, Immunologic , Female , Humans , Interleukin-1/genetics , Leukocytes, Mononuclear/drug effects , Lipopolysaccharides/pharmacology , Male , RNA, Messenger/biosynthesis , Rabbits , Radioimmunoassay , Tumor Necrosis Factor-alpha/genetics
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