Biomedical subjects

S Mihara

Publications and source records attributed to S Mihara.

At least 37 records · Page 2Linked to original sources

PubMed1999

Effects of actarit on synovial cell functions in patients with rheumatoid arthritis.

OBJECTIVE: Actarit (4-acetylaminophenylacetic acid), developed in Japan, has been shown to be effective for suppressing disease activity of rheumatoid arthritis (RA). We analyzed effects of actarit on synovial cell functions in patients with RA for insight into the clinical application of this medication. METHODS: RA primary synovial cells were co-cultured with actarit at 10(-4)-10(-7) M. Their subsequent proliferative responses and proinflammatory cytokine and matrix metalloproteinase (MMP) production at the mRNA and protein levels were measured. Effects of actarit on adhesion molecule expression were analyzed by immunofluorescence flow cytometry and cell-cell binding assay. RESULTS: Spontaneous tumor necrosis factor-alpha and interleukin 1beta secretion by primary synovial cells of patients with RA was reduced by actarit at therapeutic concentrations (10(-5)-10(-6) M). In contrast, actarit also suppressed MMP-1 production by the primary synovial cells. In addition, actarit down-regulates CD44 and intercellular adhesion molecule 1 expression on fibroblast-like synovial cell lines, and very late antigen 4 expression on CD14+ macrophage-like synovial cells resulted in the inhibition of lymphocyte adhesion to RA synovial cells. CONCLUSION: The results suggest that actarit acts on RA synovial cells to reduce cell-cell interactions with autologous synovium infiltrating lymphocytes and to inhibit proinflammatory cytokine and MMP production, leading to amelioration of symptoms of RA.

Antirheumatic Agents↗

PubMed1998

Presynaptic calcium channels mediating synaptic transmission in submucosal neurones of the guinea-pig caecum.

1. Intracellular recording techniques were used to examine the voltage-activated calcium channels mediating neurotransmitter release from nerve terminals of extrinsic, sympathetic origin and intrinsic (enteric) origin innervating submucosal neurones of the guinea-pig caecum. 2. The noradrenergic slow inhibitory postsynaptic potential (IPSP) was abolished by superfusion of omega-conotoxin (omega-CTX) GVIA (3-300 nM), with an apparent IC50 of 8.6 nM. Superfusion of omega-CTX MVIIC (500 nM) also suppressed the amplitude of slow IPSPs, but both omega-agatoxin IVA (100 nM) and nicardipine (1-10 microM) were ineffective. The hyperpolarization induced by exogenous noradrenaline was not affected by omega-CTX GVIA (100 nM). 3. In contrast to the slow IPSP, the amplitude of the cholinergic fast excitatory postsynaptic potential (EPSP) was partially inhibited, but not abolished, by omega-CTX GVIA (0.1-1 microM). Furthermore, omega-agatoxin IVA (0.1-1 microM) or omega-CTX MVIIC (0.1-1 microM) also affected the fast EPSP, but nicardipine (1-10 microM) was ineffective. In combination, omega-CTX GVIA (100 nM) and omega-agatoxin IVA (100 nM) inhibited the fast EPSP by 74 +/- 6 %; the residual fast EPSP was not affected by omega-CTX MVIIC (100 nM). The fast EPSP was completely abolished by low Ca2+, high Mg2+ Krebs solution or Krebs solution containing Co2+ (2 mM) and Cd2+ (400 microM). The depolarization induced by exogenous acetylcholine was not affected by either omega-CTX GVIA (100 nM), omega-agatoxin IVA (100 nM) or omega-CTX MVIIC (100 nM). 4. Taken together, these results suggest that, in the submucosal plexus of the guinea-pig caecum, release of noradrenaline from extrinsic nerve terminals is regulated by N-type calcium channels, whereas release of acetylcholine from intrinsic nerve terminals involves several types of calcium channel.

Acetylcholine↗

PubMed1998

Endothelin ET(B) receptors show different binding profiles in intact cells and cell membrane preparations.

We examined the affinity of endothelin-1, endothelin-3 and four endothelin receptor ligands, BQ788 (cis-2,6-dimethylpiperidinocarbonyl-gamma-methyl-Leu-D-Trp(1-CO 2CH3-D-Nle-ONa), SB-209670 ((+)-(1S,2R,3S)-3-(2-carboxymethoxy-4-methoxyphenyl)-1-(3,4-methylenedio xyphenyl)-5-(prop-1-yloxy)indane-2-carboxylic acid), IRL-1620 (succinyl-[Glu9,Ala11,15]endothelin-1(8-21)), and L-749329 (3',4'-methylenedioxy-1-(2-propyl-4-carboxyphenoxy)-N-(4-isopropyl -phenylsulfonyl)-benzene acetamide), for endothelin ET(B) receptors in human and rat heart cells. The affinities of these ligands showed good correlation between both types of living cells and between their membrane preparations (r = 0.861, P < 0.001), but less significant correlation between each of the living cells and its respective membrane preparation (r = 0.569, 0.02 < P < 0.05). These results suggest that there is no species difference in the affinities of these ligands and that destruction of the intact cell membrane structure may lead to changes in binding properties of the endothelin ET(B) receptor.

Animals↗

PubMed1998

Binding characterization of [3H]S-0139, an antagonist of the endothelin ET(A) receptor subtype.

S-0139 (27-O-3-[2-(3-carboxy-acryloylamino)-5-hydroxyphenyl]-acryloylo xy myricerone, sodium salt) is a highly specific nonpeptide endothelin ET(A) receptor antagonist. The binding of [3H]S-0139 was compared to that of [125I]endothelin-1 to characterize the binding of the antagonist in porcine aortic smooth muscle membranes. Scatchard analysis revealed a single class of [3H]S-0139 binding sites with a Kd value of 0.61 +/- 0.10 nM and a Bmax of 0.72 +/- 0.16 pmol/mg protein. These sites were saturable and reversible. [125I]Endothelin-1 also showed binding with high affinity (Kd = 0.12 +/- 0.02 nM) to a homogeneous population of binding sites, whose Bmax (0.71 +/- 0.20 pmol/mg protein) was almost the same as that for [3H]S-0139. In both cases, the binding could be displaced by known endothelin receptor ligands and their IC50 values in each case showed a very close correlation (r = 0.986). The potency of seven endothelin receptor antagonists to displace [3H]S-0139 binding also correlated highly to the potency for inhibiting the endothelin-1-induced increase in cytosolic Ca2+ concentration (r = 0.949). Myriceric acid A showed a more potent functional activity than expected from its binding affinity, but this seemed to result from the different assay conditions, such as incubation time. Together, the results suggest that S-0139 labels only endothelin ET(A) receptor binding sites in porcine aortic smooth muscle.

Animals↗

PubMed1998

Inhibition of Fas/Fas ligand-mediated apoptotic cell death of lymphocytes in vitro by circulating anti-Fas ligand autoantibodies in patients with systemic lupus erythematosus.

OBJECTIVE: The Fas/Fas ligand (FasL) system has been assigned a pivotal role in the establishment and maintenance of peripheral tolerance, and mice having defects in the Fas/FasL system are known to develop lupus-like symptoms. However, it remains unclear whether the Fas/FasL system is involved in the pathogenesis of systemic lupus erythematosus (SLE) in humans. This study examined whether there are circulating anti-FasL autoantibodies in the peripheral blood of patients with SLE that would interfere with Fas/FasL-mediated apoptosis. METHODS: Anti-FasL autoantibodies were detected by Western blot analysis using the recombinant extracellular domain of human FasL as the antigen. Apoptosis of Fas-expressing Jurkat cells, induced by recombinant soluble FasL (sFasL) in the presence of anti-FasL autoantibodies, was assessed by DNA staining with propidium iodide, followed by flow cytometric analysis. Apoptosis of Jurkat cells by cell-bound FasL was assessed by 2-color analysis, involving TUNEL staining with fluorescein isothiocyanate-dUTP and phycoerythrin-labeled anti-CD3 monoclonal antibodies. RESULTS: Among the 21 patients with SLE, 7 had IgG-isotype anti-FasL autoantibodies in their circulating blood. In addition, these autoantibodies inhibited both sFasL-mediated and cell-bound FasL-mediated apoptosis of Fas-expressing Jurkat cells. Thus, it is plausible that anti-FasL autoantibodies in patients with SLE disturb the establishment and maintenance of peripheral tolerance in vivo by inhibiting the Fas/FasL-mediated elimination of autoreactive lymphocytes. CONCLUSION: These results suggest that anti-FasL autoantibodies that inhibit Fas/FasL-mediated apoptosis are involved, at least in part, in immune abnormalities and may possibly be involved in the pathogenesis of SLE.

Adolescent↗

PubMed1998

Mild hypothermia protects rat hippocampal CA1 neurons from irreversible membrane dysfunction induced by experimental ischemia.

In order to examine the effects of hypothermia on the changes in membrane potential induced by experimental ischemia (deprivation of oxygen and glucose), intracellular recordings were made from single CA1 pyramidal neurons in slice preparations of rat hippocampus. Application of ischemic medium caused irreversible changes in membrane potential consisting of an initial hyperpolarization, then a slow depolarization and a rapid depolarization. At temperatures of 35 degrees C and 37 degrees C, once the rapid depolarization occurred, readministration of oxygen and glucose failed to restore the membrane potential, a state referred to as irreversible membrane dysfunction. When the temperature was lowered to between 27 degrees C and 33 degrees C, the membrane potential returned to the control resting membrane potential in 75% of the neurons. The temperature coefficients (Q10) of the latency, the amplitude, and the maximal slope of the rapid depolarization were 2.5, 1.4 and 2.9, respectively. It is concluded that the critical neuroprotective temperature in ischemia-induced membrane dysfunction is found to be 33 degrees C in single CA1 neurons in vitro.

Animals↗

PubMed1998

Nitric oxide contributes to irreversible membrane dysfunction caused by experimental ischemia in rat hippocampal CA1 neurons.

The effects of agents which affect the action of nitric oxide (NO) were studied intracellularly on the ischemia-induced changes in membrane potential of single CA1 pyramidal neurons of the rat hippocampal slice preparations. The N-methyl-D-aspartate (NMDA) receptor antagonists, (+/-)-2-amino-5-phosphonopentanoic acid (AP5, 250 microM) or Co2 (2 mM) restored the membrane potential in more than 80% of the neurons. In about 60% of the neurons, the membrane potential was partially recovered as a result of exposure to the NO synthase inhibitor, NG-nitro-L-arginine (100 microM). The NO scavengers, carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO, 300 microM) and hemoglobin (10 microM) restored the membrane potential in all neurons examined. Superoxide dismutase (50 U/ml) protected about 75% of the neurons from irreversible membrane dysfunction. It is concluded that the release of NO induced by experimental ischemia may result in the irreversible membrane dysfunction, and that a NO scavenger, carboxy-PTIO, prevents the ischemic changes in membrane potential. With respect to ischemic brain damage, the neuroprotection provided by carboxy-PTIO may have clinical relevance in the management of a variety of neurological conditions.

2-Amino-5-phosphonovalerate↗

PubMed1998

Efficacy of ultrasonic mass survey for abdominal cancer.

From August 1983 through March 1995, 204,099 people received ultrasonic mass survey of the abdomen for the first time. Among these examinees, 631 (0.31%) malignant neoplasm cases, such as 201 hepatocellular carcinoma (HCC), 81 gallbladder (GB) cancer, 57 pancreatic cancer, and 169 renal cell carcinoma (RCC), were detected. Three hundred seventy six out of 590 cases (64%), excluding chronic leukemia cases and metastatic liver cancer cases, were surgically resected. The resection rate of HCC, GB cancer, pancreatic cancer, and RCC were 25%, 88%, 49%, and 99%, respectively. The cumulative survival rate of the 376 resected cases was 79.5% at 10 years. The cumulative survival rates of resected cases of HCC, GB cancer, pancreatic cancer and cumulative survival rates of resected cases of HCC, GB cancer, pancreatic cancer and RCC were 34% at ten years, 83% at 10 years, 49% at 7 years, and 99% at 10 years, respectively. Ultrasonic mass survey is dramatically useful for early detection of various kinds of abdominal cancers, especially RCC and GB cancer. From now on, many earlier abdominal cancers will be found by establishing and promoting ultrasonic mass survey systems.

Abdominal Neoplasms↗

PubMed1998

Zinc ions prevent processing of caspase-3 during apoptosis induced by geranylgeraniol in HL-60 cells.

Geranylgeraniol (GGO) at 50 microM induces apoptosis in HL-60 cells. We examined the effects of Zn2+ ions on this process. Treatment of HL-60 cells with Zn2+ ions inhibited subsequent GGO-induced fragmentation of DNA. In a cell-free system that consisted of a specific substrate for caspase-3 and a lysate of HL-60 cells that had been treated with 50 microM GGO, Zn2+ ions at concentrations above 0.1 mM inhibited the activity of caspase-3. The effect of Zn2+ ions on the processing of caspase-3 during GGO-induced apoptosis was investigated by Western blotting, which revealed that an inactive 32-kDa precursor of caspase-3 was cleaved, in response to GGO, to yield an activated 17-kDa enzyme. Treatment of HL-60 cells with Zn2+ ions inhibited the cleavage of the precursor by a protease that was induced by treatment with GGO, and inhibition of this processing was well correlated with the inhibition by Zn2+ ions of caspase-3 activity in the cell-free system. In cell-extracted cytosols, Zn2+ ions inhibited the cleavage of the 32-kDa precursor by caspase-9 (Aapf-3) that was activated by addition of cytochrome c and dATP. These results indicate that inhibition of GGO-induced apoptosis in HL-60 cells by Zn2+ ions might be due to inhibition by Zn2+ ions of the processing of a precursor to caspase-3.

Apoptosis↗

PubMed1998

[Kidney-sparing surgery for recurrent ureteral and bladder cancers in an aged patient with functionally solitary horseshoe kidney].

A 79-year-old woman was admitted with recurrent ureteral and bladder cancers. She had a horseshoe kidney with a non-functioning right renal unit. Fifteen months earlier, multiple urothelial tumors had first developed in the left upper ureter and bladder. Transurethral resection of bladder tumor (TUR-Bt) and partial ureterectomy (2 cm) had been performed. Presently, the recurrent tumors were located at the left lower ureter and bladder. Considering the high age of the patient, TUR-Bt and partial ureterectomy (5 cm) were performed. Besides urothelial cancers, she had been operated for carcinomas of the colon, uterus and stomach. Kidney-sparing therapy has successfully maintained her quality of life.

Aged↗

PubMed1998

Prevalence and conditions of urinary incontinence among the elderly.

In Japan, elderly disorders and diseases have markedly increased in recent years, because of rapid aging and an increasing number of older persons. The situation is creating serious social and community problems. These disorders, particularly dysuria and urinary incontinence (UI), disturb the quality of life (QOL) in latelife. Few reports on UI have been published, but precise investigation into the community level remains to be made. Our presentation is the development, implementation and evaluation of elderly UI in Kumamoto Prefecture. This study includes 2,304 people (male: 856, female: 1,448), over 65 years of age, living in two different communities; one is an urban (K) and the other is a typical rural area (S). The rate of UI was in homebound elderly persons, male: 4.7%, female: 11.3%, and in nursing home residents, male: 16.2%, female: 23.2%. The condition of UI was: almost Urinary Urgency in male (61.5%), and Stress Incontinence (such as, caused by coughing, sneezing, and exercise) in female (46.3%). The influence of UI on the activity of daily life was investigated. Most of the male cases were giving concerns for family and community. In contrast, females hesitated to participate in group excursions and outdoor exercise, and had a tendency to live alone or indoors. However, most persons (81.5%) with UI did not visit a physician. From this investigation, we conclude that a community health care program and public support system are essential for proper understanding and solution of the elderly UI problem.

Activities of Daily Living↗

PubMed1997

In vivo mechanisms for the inhibition of T lymphocyte activation by long-term therapy with tacrolimus (FK-506): experience in patients with Behçet's disease.

OBJECTIVE: To examine the in vivo mechanisms of suppression of T lymphocyte function in patients with Behçet's disease (BD) undergoing long-term treatment with tacrolimus (FK-506). METHODS: Intracellular proteins were analyzed by immunoprecipitation and Western blotting. Messenger RNA expression was studied by a polymerase chain reaction-based technique. RESULTS: Interleukin-2 production was suppressed in patients treated with tacrolimus. This suppression was found to be due to inhibition of interactions between activated calcineurin (Cn) and nuclear factor of activated T cells (NF-AT), inhibition of cleavage of the autoinhibitory domain of the CnA subunit, and inhibition of heterodimer formation by CnA and CnB subunits, resulting in the absence of NF-AT in nuclei of the T cells. We found that T lymphocytes in some BD patients treated with tacrolimus had reduced amounts of FK-506 binding protein (FKBP) in their cytoplasm. CONCLUSION: Tacrolimus reduces the Cn activity of T cells in vivo by the cumulative effects of several distinct mechanisms. It is plausible that reduced amounts of FKBP may be associated with diminished clinical efficacy in some BD patients receiving prolonged treatment with tacrolimus.

Adult↗

PubMed1997

Electrophysiology of neurochemically identified submucosal neurones of the guinea-pig intestine.

A number of electrophysiological studies have shown that neurones in the submucous plexus are endowed with three major types of synaptic potentials in response to nerve stimulation: a fast EPSP, a slow IPSP, and a slow EPSP. Combined electrophysiological and immunohistochemical studies enabled analysis of the types of neurochemically identified neurones which receive each type of synaptic input. This short review briefly summarizes the results obtained from these studies.

Animals↗

PubMed1997

Development of pathogenic anti-DNA antibodies in patients with systemic lupus erythematosus.

The anti-DNA response is a hallmark of systemic lupus erythematosus (SLE). The precise mechanisms leading to anti-DNA antibody (Ab) production remain to be studied. Nonetheless, it is becoming clear that anti-DNA Abs cause inflammatory lesions not only via deposition of circulating immune complexes (IC) consisting of anti-DNA Ab and antigens (Ags), but also via in situ IC formation by cationic anti-DNA Abs. It is intriguing that cationic anti-DNA Abs are encoded by a unique germline Vkappa gene, A30, which encodes an extraordinary cationic light chain, whereas somatic mutations did not induce a cationic shift of electrical charge in human lupus nephritis, suggesting that the usage of a specific germline gene may confer the cationic charge (or pathogenicity) on anti-DNA Abs and that somatic mutations induce the affinity maturation of Abs. Whether cationic anti-DNA Abs will develop depends at least partly on the presence or absence of the germline A30 gene, since patients who lack this gene in the germline Vkappa repertoire did not develop severe lupus nephritis. Receptor editing, a mechanism for changing the affinity of the B cell Ag receptor [surface immunoglobulin (Ig) receptor] to avoid self-reactivity actually seems defective in patients with SLE because normal B cells edited the A30 gene, whereas SLE B cells express A30 mRNA. Thus, along with the importance of somatic mutations, polymorphisms of Ig Vkappa locus, and genetic predisposition, the failure of receptor editing may contribute to the development of pathogenic anti-DNA responses in humans.

Antibodies, Antinuclear↗

PubMed1997

Electrophysiological characteristics of submucosal neurones in the proximal colon of guinea-pigs: comparisons with caecum and descending colon.

A systematic examination has been made of the active and passive electrophysiological properties and synaptic inputs of forty-four randomly impaled submucosal neurones in the proximal colon of the guinea-pig to compare these characteristics directly with those of submucosal neurones in the caecum (n = 70) and descending colon (n = 45). Within each of the three electrophysiological classes of submucosal neurones identified (S, S/AH and AH), no statistically significant regional differences were found with respect to the resting membrane potential, membrane time constant or input resistance between neurones of the proximal colon, descending colon and caecum. Of submucosal neurones from the proximal colon, forty-three of forty-four (98%) received fast excitatory synaptic potentials (fast EPSPs); thirty-nine (91%) were S neurones and the others were S/AH neurones; only one of the forty-four cells (2%) was an AH neurone. An idazoxan-sensitive slow inhibitory postsynaptic potential (slow IPSP) was induced in thirty of forty-three S and S/AH neurones (70%) of the proximal colon, compared with sixty-one of sixty-six caecal neurones (92%) and twelve of forty-one neurones (29%) in the descending colon. The mean (+/- S.E.M.) amplitude of the slow IPSP in proximal colonic neurones was 17 +/- 1 mV (range, 6-30 mV; n = 30), compared with the significantly larger synaptic response (25 +/- 1 mV; range, 7-38 mV; n = 66; P < 0.05) recorded in the caecum; the mean slow IPSP amplitude in the descending colon was significantly smaller (12 +/- 2 mV; range, 5-27 mV; n = 12; P < 0.05) than that in the caecum. In the proximal colon and caecum, only those neurones with a slow IPSP had a hyperpolarizing response to noradrenaline, whereas about 50% of those neurons of the descending colon that lacked a slow IPSP were hyperpolarized by noradrenaline, acting via alpha 2-adrenoceptors. Thus, the electrophysiological characteristics of the submucosal neurones of the proximal colon more closely resemble those of the caecum than those of the descending colon, of which many do not have a functional noradrenergic synaptic input. Furthermore, the results confirm that there are fundamental regional differences in the guinea-pig large intestine with respect to the synaptic organization of submucosal neurones of particular electrophysiological classes.

Adrenergic alpha-Agonists↗

PubMed1997

Mechanisms underlying the rapid depolarization produced by deprivation of oxygen and glucose in rat hippocampal CA1 neurons in vitro.

Intracellular recordings were made to investigate the mechanism, site, and ionic basis of generation of the rapid depolarization induced by superfusion with ischemia-simulating medium in hippocampal CA1 pyramidal neurons of rat tissue slices. Superfusion with ischemia-simulating medium produced a rapid depolarization after approximately 6 min of exposure. When oxygen and glucose were reintroduced, the membrane potential did not repolarize but depolarized further, reaching 0 mV approximately 5 min after reintroduction. Simultaneous recordings of changes in cytoplasmic Ca2+ concentration ([Ca2+]i) and membrane potential recorded from 1-[6-amino-2-(5-carboxy-2-oxazolyl)-5-benzofuranyloxy]-2-(2- amino-5-methylphenoxy)-ethane-N,N,N',N'-tetraacetic acid pentaacetoxymethyl ester (Fura-2/AM) loaded slices revealed a rapid increase in [Ca2+]i in all CA1 layers corresponding to the rapid depolarization of the soma membrane. The result suggests that the rapid depolarization is generated not only in the soma but also in the apical and basal dendrites. Application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), DL-2-amino-4-phosphonobutyric acid, and DL-2-amino-3-phosphonopropionic acid or bicuculline did not affect the amplitude and the maximal slope. Reduction in the concentration of extracellular Ca2+ or addition of CNQX or DL-2-amino-5-phosphonopentanoic acid delayed the onset of the rapid depolarization. The amplitude of the rapid depolarization recorded with Cs acetate electrodes in tetraethylammonium-containing medium had a linear relationship to the membrane potential between -50 and 20 mV. The reversal potential was shifted in the hyperpolarizing direction by a decrease in either [Na+]o or [Ca2+]o, whereas the reversal potential was shifted in the depolarizing direction by a decrease in [Cl-]o or using CsCl electrodes. An increase or decrease in [K+]o did not affect the reversal potential. These results indicate that the rapid depolarization is Na+, Ca2+, and Cl- dependent. The lack of effects of changes in [K+]o is probably due to the accumulation of interstitial K+ before generating the rapid depolarization. Prolonged application of ouabain (30 microM) caused an initial small hyperpolarization, a subsequent slow depolarization, and a rapid depolarization. In summary, the present study has demonstrated that the rapid depolarization is voltage-independent and is probably due to a nonselective increase in permeability to all participating ions, which may occur only in pathological conditions. The underlying conductance change is primarily the result of inhibition of Na,K-ATPase activity in the recorded neuron.

Animals↗

PubMed1997

Mechanisms of KE298, 2-acetylthiomethyl-3-(4-methylbenzoyl) propionic acid, to suppress abnormal synovial cell functions in patients with rheumatoid arthritis.

OBJECTIVE: 2-Acetylthiomethyl-3-(4-methylbenzoyl) propionic acid, KE298, a derivative or propionic acid developed in Japan has been shown to be effective for suppressing disease activity of rheumatoid arthritis (RA) in clinical trials in Japan. It is thus a candidate as a new disease modifying antirheumatic drug (DMARD). We analyzed effects of KE298 on synovial fibroblast-like cells in patients with RA to obtain insight into the clinical application of this medication. METHODS: RA synovial fibroblast-like cells were co-cultured with KE298 at 10(-4)-10(-5) M in the presence or absence of tumor necrosis factor-alpha 2 ng/ml, and their subsequent proliferative responses and proinflammatory cytokine and matrix metalloproteinase (MMP) production at the mRNA and protein levels were measured. Effects of KE298 on MMP-1 gene transcription and AP-1 transcription factor expression of RA synovial cells were studied by chloramphenicol acetyltransferase assay and gel shift assay, respectively. RESULTS: KE298 inhibited proliferation of RA synovial cells, proinflammatory cytokine production, and MMP-1 production mainly by reducing their transcription via downmodulation of AP-1 transcription factor. CONCLUSION: KE298 inhibits aberrant synovial cell functions of patients with RA by downregulating gene transcription, suggesting clinical application and usefulness of this new DMARD.

Antirheumatic Agents↗

PubMed1996

Characterization of a germline Vk gene encoding cationic anti-DNA antibody and role of receptor editing for development of the autoantibody in patients with systemic lupus erythematosus.

We found previously that cationic anti-DNA autoantibodies (autoAbs) have nephritogenic potential and usage of a specific germline Vk gene, A30, has major influences on cationic charge of the autoAb in human lupus nephritis. In the present study, we have characterized A30 germline Vk gene using cosmid cloning technique in patients with SLE. A30 gene locus locates in less than 250 kb from the Ck region, and the cationic anti-DNA mRNA used the upstream Jk2 gene, indicating that cationic anti-DNA mRNA is a product of primary gene rearrangement. By using PCR technique, we found that A30 gene locus in the genome was defective in eight out of nine SLE patients without nephritis. In contrast, all nine patients with lupus nephritis had intact A30 gene. The presence and absence of A30 gene was associated with the development of lupus nephritis or not (P < 0.01, by Fisher's exact test, two-sided). It was thus suggested that absence of functional A30 gene may rescue from developing lupus nephritis in the patients. A30 is reported to be a potentially functional but rarely expressed Vk gene in humans. It is possible that normal B cells edit primarily rearranged A30 gene with autoreactive potentials by receptor editing mechanism for changing the affinity of the B cell Ag receptor to avoid self-reactivity, whereas SLE B cells may have a defect in this mechanism. Indeed, we found that normal B cells edit A30-Jk2 gene in their genome possibly by inversion mechanism, whereas SLE B cells contain rearranged A30-Jk2-Ck gene in the genome and express A30-associated mRNA, suggesting that receptor editing mechanism is also defective in patients with SLE. Our study suggests that polymorphism of Ig Vk locus, and failure of receptor editing may contribute to the development of pathogenic anti-DNA responses in humans.

Adolescent↗