Biomedical subjects

X Meng

Publications and source records attributed to X Meng.

At least 109 records · Page 6Linked to original sources

PubMed1999

Tumor necrosis factor-alpha and interleukin-1beta synergistically depress human myocardial function.

OBJECTIVE: Proinflammatory cytokines such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta have been implicated in the pathogenesis of myocardial dysfunction in ischemia-reperfusion injury, sepsis, chronic heart failure, viral myocarditis, and cardiac allograft rejection. Although circulating TNF-alpha and IL-1beta are both often elevated in septic shock, it remains unknown whether TNF-alpha or IL-1beta are the factors induced during sepsis that directly depress human myocardial function, and if so, whether the combination synergistically depresses myocardial function. Furthermore, the mechanism(s) by which these cytokines induce human myocardial depression remain unknown. We hypothesized the following: a) TNF-alpha and IL-1beta directly depress human myocardial function; b) together, TNF-alpha and IL-1beta act synergistically to depress human myocardial function; and c) inhibition of ceramidase or nitric oxide synthase attenuates myocardial depression induced by TNF-alpha or IL-1beta by limiting proximal cytokine signaling or production of myocardial nitric oxide (NO). DESIGN: Prospective, randomized, controlled study. SETTING: Experimental laboratory in a university hospital. SUBJECTS: Freshly obtained human myocardial trabeculae. INTERVENTIONS: Human atrial trabeculae were obtained at the time of cardiac surgery, suspended in organ baths, and field simulated at 1 Hz, and the developed force was recorded. After a 90-min equilibration, TNF-alpha (1.25, 12.5, 125, or 250 pg/mL for 20 mins), IL-1beta (6.25, 12.5, 50, or 200 pg/mL for 20 mins), or TNF-alpha (1.25 pg/mL) plus IL-1beta (6.25 pg/mL) were added to the bath, and function was measured for the subsequent 100 mins after the 20-min exposure. To assess the roles of the sphingomyelin and NO pathways in TNF-alpha and IL-1beta cross-signaling, the ceramidase inhibitor N-oleoyl ethanolamine (1 microM) or the NO synthase inhibitor N(G)-monomethyl-L-arginine (10 microM) was added before TNF-alpha (125 pg/mL) or IL-1beta (50 pg/mL). MEASUREMENTS AND MAIN RESULTS: TNF-alpha and IL-1beta each depressed human myocardial function in a dose-dependent fashion (maximally depressing to 16.2 + 1.9% baseline developed force for TNF-alpha and 25.7 + 6.3% baseline developed force for IL-1beta), affecting systolic relatively more than diastolic performance (each p < .05). However, when combined, TNF-alpha and IL-1beta at concentrations that did not individually result in depression (p > .05 vs. control) resulted in contractile depression (p < .05 vs. control). Inhibition of myocardial sphingosine or NO release abolished the myocardial depressive effects of either TNF-alpha or IL-1beta. CONCLUSIONS: TNF-alpha and IL-1beta separately and synergistically depress human myocardial function. Sphingosine likely participates in the TNF-alpha and IL-1beta signal leading to human myocardial functional depression. Therapeutic strategies to reduce production or signaling of either TNF-alpha or IL-1beta may limit myocardial dysfunction in sepsis.

Amidohydrolases↗

PubMed1999

Reduction of infarct size in the rat heart by LPS preconditioning is associated with expression of angiogenic growth factors and increased capillary density.

Inflammation induces the expression of angiogenic growth factors in tissues, which leads to microvascular growth. Bacterial lipopolysaccharide (LPS) provokes a transient inflammatory response in the heart and induces delayed cardiac resistance to post-ischemic contractile dysfunction. In this study, we examined: 1) the effects of LPS on myocardial expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), 2) whether an increase in the density of myocardial microvessels follows the expression of angiogenic growth factors, and 3) the effect of LPS on myocardial resistance to infarction and its relationship with microvascular growth. Rats were treated with LPS (from Salmonella typhimurium, 0.5 mg/kg i.p.). The expression of bFGF and VEGF in the myocardium was examined at 6 and 12 h after LPS treatment by immunofluorescent staining. Myocardial capillary and arteriole densities were determined 3 days after LPS treatment by morphometry, using immunofluorescent staining of von Willebrand factor (a marker protein of endothelial cells) and alpha-smooth muscle actin (a marker protein of smooth muscle cells). To examine cardiac resistance to infarction, hearts were subjected to 40 min of regional ischemia and 2 h of reperfusion by reversible occlusion of left coronary artery at 3 days after LPS treatment. LPS induced cardiac bFGF and VEGF at 6 and 12 h after treatment. The expression of these growth factors was followed by an increase in myocardial capillary density (2032 +/- 78/mm2 vs. 1617 +/- 47/mm2 in saline control, P < 0.05), but not arteriole density, at 3 days. Meanwhile, infarct size was significantly reduced by LPS preconditioning (infarct/left ventricle 12.3 +/- 1.04% vs. 21.7 +/- 1.65% in saline control, 43% reduction, P < 0.05). These results suggest that LPS preconditioning induces cardiac bFGF and VEGF, and an increase in myocardial capillary density. This increased myocardial capillary density is associated with a reduced infarct size after in vivo regional ischemia-reperfusion.

Animals↗

PubMed1999

Isolation and characterisation of neurotoxigenic Clostridium butyricum from soil in China.

Soil specimens collected from a site around the home of patients with food-borne type E. botulism probably caused by neurotoxigenic Clostridium butyricum in Guanyun, Jiangsu province, China, were examined for the presence of neurotoxigenic C. butyricum. Five lakeside sites of Weishan lake, in an area near to the sites where the type E. botulism outbreaks caused by neurotoxigenic C. butyricum occurred were also surveyed. Type E toxin-producing C. butyricum was isolated from soil from four sites including the site in Guanyun. Polymerase chain reaction assay demonstrated the presence of the type E toxin gene in all the toxigenic isolates. The biochemical properties of the isolates from the Guanyun soil and the lakeside soil were identical except for inulin fermentation and starch hydrolysis properties. These results indicate that neurotoxigenic C. butyricum has its principal habitat in soil.

Bacterial Toxins↗

PubMed1999

Adrenergic induction of bimodal myocardial protection: signal transduction and cardiac gene reprogramming.

This study tested the hypothesis that in vivo norepinephrine (NE) treatment induces bimodal cardiac functional protection against ischemia and examined the roles of alpha1-adrenoceptors, protein kinase C (PKC), and cardiac gene expression in cardiac protection. Rats were treated with NE (25 micrograms/kg iv). Cardiac functional resistance to ischemia-reperfusion (25/40 min) injury was examined 30 min and 1, 4, and 24 h after NE treatment with the Langendorff technique, and effects of alpha1-adrenoceptor antagonism and PKC inhibition on the protection were determined. Northern analysis was performed to examine cardiac expression of mRNAs encoding alpha-actin and myosin heavy chain (MHC) isoforms. Immunofluorescent staining was performed to localize PKC-betaI in the ventricular myocardium. NE treatment improved postischemic functional recovery at 30 min, 4 h, and 24 h but not at 1 h. Pretreatment with prazosin or chelerythrine abolished both the early adaptive response at 30 min and the delayed adaptive response at 24 h. NE treatment induced intranuclear translocation of PKC-betaI in cardiac myocytes at 10 min and increased skeletal alpha-actin and beta-MHC mRNAs in the myocardium at 4-24 h. These results demonstrate that in vivo NE treatment induces bimodal myocardial functional adaptation to ischemia in a rat model. alpha1-Adrenoceptors and PKC appear to be involved in signal transduction for inducing both the early and delayed adaptive responses. The delayed adaptive response is associated with the expression of cardiac genes encoding fetal contractile proteins, and PKC-betaI may transduce the signal for reprogramming of cardiac gene expression.

Actins↗

PubMed1999

Early kidney TNF-alpha expression mediates neutrophil infiltration and injury after renal ischemia-reperfusion.

The purpose of this study was to determine whether isolated renal ischemia and reperfusion (I/R) induces renal tumor necrosis factor (TNF) mRNA production, TNF protein expression, or TNF bioactivity and, if so, whether local/early TNF production acts as mediator of ischemia-induced, neutrophil-mediated renal injury. After rats were anesthetized, varying periods of renal ischemia, with or without reperfusion, were induced. Kidney mRNA content (RT-PCR), TNF protein expression (ELISA), TNF bioactivity (WEHI-164 cell clone cytotoxicity assay), and neutrophil infiltration [myeloperoxidase (MPO) assay] were determined. In other animals, renal MPO and serum creatinine were assessed after TNF was neutralized [binding protein (TNF-BP)]. Thirty minutes of ischemia induced renal TNF mRNA. TNF protein expression and bioactivity peaked after 1 h ischemia and 2 h reperfusion, whereas neutrophil infiltration peaked at 4 h reperfusion. TNF-BP neutralized TNF bioactivity, reduced neutrophil infiltration, and protected postischemic function. These results constitute the initial demonstration that 1) early renal tissue TNF expression contributes to neutrophil infiltration and injury after I/R and 2) TNF-BP may offer a new adjunctive therapy in renal preservation prior to planned ischemic insults.

Animals↗

PubMed1999

Attenuation of renal ischemia-reperfusion injury in inducible nitric oxide synthase knockout mice.

Renal ischemia-reperfusion (I/R) injury was investigated in inducible nitric oxide synthase (iNOS) knockout mice. After a 26-min bilateral renal pedicle clamp, serum creatinine concentrations (in mg/dl) in wild-type mice after a 24-h reperfusion were 0.25 +/- 0.03 in sham-operated controls and 2.3 +/- 0.38 in ischemic mice (P < 0. 01); after 48 h, concentrations (in mg/dl) were 0.25 +/- 0.03 in controls and 2.0 +/- 0.18 in ischemic mice (P < 0.01). iNOS knockout mice demonstrated an attenuation of serum creatinine concentration after renal I/R injury. Serum creatinine concentrations (mg/dl) after a 24-h reperfusion were 2.3 +/- 0.22 in wild-type ischemic and 1.21 +/- 0.25 in iNOS knockout ischemic mice (P < 0.05); after 48 h, concentrations were 2.0 +/- 0.18 in wild-type ischemic and 0.96 +/- 0.25 in iNOS knockout ischemic mice (P < 0.01). Histological scoring of acute tubular necrosis in iNOS knockout mice was decreased compared with that in wild-type controls (0.88 +/- 0.2 vs. 3.3 +/- 0. 3, P < 0.05). iNOS protein in the renal cortex of wild-type mice subjected to renal I/R injury was undetectable up to 48 h. However, a strong upregulation of heat shock protein 72 expression was observed in renal cortex of iNOS knockout mice under basal conditions. In conclusion, kidneys of iNOS knockout mice were protected against ischemic acute renal failure. This protective effect may be related to a compensatory upregulation of heat shock protein 72.

Animals↗

PubMed1999

Inhibition of PARP prevents oxidant-induced necrosis but not apoptosis in LLC-PK1 cells.

Oxidant-induced cell injury has been implicated in the pathogenesis of several forms of acute renal failure. The present studies examined whether activation of poly(ADP-ribose)polymerase (PARP) by oxidant-induced DNA damage contributes to oxidant injury of renal epithelial cells. H2O2 exposure resulted in an increase in PARP activity and decreases in cell ATP and NAD content. These changes were significantly inhibited by 10 mM 3-aminobenzamide (3-ABA), a PARP inhibitor. In contrast, H2O2-induced DNA damage was not prevented by 3-ABA. Exposure of LLC-PK(1) cells to 1 mM H2O2 for 2 h induced necrotic cell death as measured by increased lactate dehydrogenase (LDH) release. 3-ABA completely prevented the H2O2-induced LDH release. Live/dead fluorescent staining confirmed the protection by 3-ABA. These results are consistent with the view that oxidant-induced DNA damage activates PARP and that the subsequent ATP and NAD depletion contribute to necrotic cell death. Of note, although protected from necrosis, cells treated with H2O2 and 3-ABA underwent apoptosis as evidenced by DNA fragmentation and bis-benzimide staining. In conclusion, activation of PARP contributes to oxidant-induced ATP depletion and necrosis in LLC-PK1 cells. However, PARP inhibition may target cells toward an apoptotic form of cell death.

Animals↗

PubMed1999

Liposomal delivery of heat-shock protein 72 into the heart prevents endotoxin-induced myocardial contractile dysfunction.

BACKGROUND: The purposes of this study were to (1) determine whether functional heat-shock protein 72 (HSP-72) may be delivered into the heart, (2) determine whether HSP-72 itself is protective against endotoxin (lipopolysaccharide [LPS]-induced cardiodepression, and (3) compare relative protection and time courses required for protection for thermally induced HSP-72 versus liposomally introduced HSP-72. METHODS: HSP-72 was introduced (liposomal HSP-72) or induced (heat shock, 42 degrees C x 15 minutes, 24 hours before) in rat heart before LPS administration (0.5 mg/kg intraperitoneal or ex vivo coronary infusion). Western blot analysis for HSP-72 was used to confirm its expression. Left ventricular developed pressure (Langendorff) was used as an index of cardiac function. RESULTS: Direct intracoronary perfusion of liposomal HSP-72 delivered functioning HSP-72 into the myocardium. LPS induced cardiodepression; however, heat shock pretreatment abolished LPS-induced contractile dysfunction. A direct connection was found between HSP-72 and protection derived from liposomal transfer experiments that similarly reduced LPS-induced cardiodepression. CONCLUSIONS: (1) HSP-72 prevents LPS-induced myocardial contractile dysfunction, (2) liposomal transfer of HSP-72 into the myocardium provides the first direct mechanistic connection between myocardial HSP-72 and protection against LPS, (3) HSP-72 induction requires 24 hours and liposomal transfer of HSP-72 requires 90 minutes, and (4) HSP-72 may offer a clinically acceptable means of protecting the heart.

Animals↗

PubMed1999

[Relationship between methylation status of gamma-glutamyl transpeptidase (GGT) genes and abnormal expression of its enzyme proteins in tissues of human hepatomas].

OBJECTIVE: To explore the mechanism of expression and alteration of gamma-glutamyl transpeptidase (GGT) genes during the development of human hepatomas. METHODS: The total GGT protein and total RNA were purified in human hepatomas, adjacent paracancerous and distal cancerous tissues. The specific activities of total GGT, membrane-combine GGT and soluble GGT were investigated, the GGT gene of 5'-NC region was amplified by using a nest RT-PCR assay, and methylation status of GGT gene M3 site were analyzed in the present study. RESULTS: An increasing tendency (P < 0.05) of total RNA concentrations was found from cancer in distal cancerous tissues; the specific activities (U/g) of total GGT, membrane-combine GGT and soluble GGT were significantly higher (P < 0.05) in hepatomas than those in adjacent paracancerous or and that distal cancerous tissues; and the frequencies of amplified fragment and hypomethylated M3 site of GGT 5'-NC region genes were 100% and 75% in hepatomas, 85% and 55% in adjacent cancerous, and 75% and 50% in distal cancerous tissues, respectively. CONCLUSIONS: The present data suggest that the abnormal expression of GGT proteins in hepatomas was related to hypomethylation status of GGT genes, and that the fragment analysis of the GGT genes might be a sensitive assay to monitor the hepatic cell canceration.

Carcinoma, Hepatocellular↗

PubMed1999

[Injury to urinary system in the operation of gynecology].

OBJECTIVE: To reduce the incidence of urinary system injury from gynecology operations. METHODS: 26 cases with urinary system injuries during gynecological operations were analyzed from 1982 to 1996. RESULTS: The incidence of urinary injury in this hospital was 0.34%, and injuries occurred most frequently in the bladder. Among the 17 cases of bladder injuries, 12 occurred during tubal ligation, 3 during cesarean section, and 2 in myomectomy. There were 9 cases of ureter injuries, 6 cases occurred in radical hysterectomy for cervical cancer, 1 in cesarean section with hysterectomy, and 2 in hysterectomy with broad ligament leiomyoma. CONCLUSIONS: This study demonstrated that it was important to be familiar with the regional anatomy, to operate strictly and separate those organs carefully in preventing the urinary system injury.

Adult↗

PubMed1999

[Estimation and clinical significance of serum soluble tumor necrosis factor receptors in patients with acute leukemia].

OBJECTIVE: To explore the levels of serum soluble tumor necrosis factor receptors (sTNFRs) and the relationships with clinical situation, chemotherapeutic effects and TNF alpha level in acute leukemia (AL) patients. METHODS: Serum levels of sTNFRs and TNF alpha were measured by ELISA. Six parameters which might influence therapeutic effects were analyzed by logistic regression. RESULTS: 1. Serum levels of sTNFR I and sTNFR II in 31 untreated AL patients were significantly higher than those in controls(P < 0.001 and < 0.005, respectively) and in bone marrow remission (BMR) AL patients (P < 0.005 and < 0.05, respectively). The levels of sTNFR I and sTNFR II in 11 relapsed/refractory AL patients were significantly higher than those in controls(P < 0.001 and < 0.05, respectively), but were not different from those in untreated AL patients. 2. Serum levels of sTNFR I in untreated ALL patients were significantly higher than those in ANLL patients (P < 0.05). 3. High levels of serum sTNFR I were more common in patients with high leukocyte count(> or = 100 x 10(9)/L) and high levels of serum sTNFR II were more common in patients with splenomegaly. Levels of the two sTNFRs positively correlated with blasts in peripheral blood (P < 0.001 and < 0.05, respectively). 4. The level of TNF alpha in 31 untreated AL patients significantly correlated with the level of sTNFR I (r = 0.440, P < 0.05). 5. The BMR rate was higher in those serum sTNFR I level < 2.0 micrograms/L than in those > or = 2.0 micrograms/L(P < 0.05). 6. Of the six parameters which might influence AL therapeutic effects, only sTNFR I < 2.0 micrograms/L was predictive. CONCLUSION: High levels of serum sTNFRs in AL patients might be derived from leukemic cells and predicted an adverse prognosis.

Adolescent↗

PubMed1999

[Cloning of differentiation-related genes induced by all-trans retinoic acid(ATRA) from human lung cancer GLC-82 cell line].

OBJECTIVE: Cloning of differentiation-related genes induced by all-trans retinoic acid(ATRA) from human lung cancer GLC-82 cell line. METHODS: The difference in gene expression between human lung cancer cell GLC-82 before and 8, 24 hours and 4 days after ATRA treatment was investigated by an improved protocol of mRNA differential display with SYBR Green I, a highly sensitive fluorescent stain. RESULTS: There were obvious differences in gene expression between human lung cancer cells before and after ATRA treatment. The expression of some genes was decreased or inhibited while that of other genes was activated temporarily or persistently. Three cDNA fragments of the differentially expressed genes were cloned and analyzed. CONCLUSION: ATRA plays an important role in regulating the differentiation-related gene expression. ATRA-induced differentiation is a complex process with multiple genes involved.

Base Sequence↗

PubMed1999

[Effects of combined treatment with flouride and 1,25-dihydroxyvitamin D3 on the histomorphometry and biomechanical properties of bone in ovariectomized rats].

OBJECTIVE: To evaluate the potential use of a combination of bone formation-promoting agents and a differentiation agent of osteoblast as a treatment for postmenopausal osteoporosis. METHODS: The effects of combined and separate administration of flouride-Ca (0.45 ng F- + 13.56 mg Ca2+)/(kg.d) and 1,25-dihydroxyvitamin D (135 pmol/d s.c.) on the histomorphometric parameters in the secondary spongiosa of proximal tibia and biomechanical properties of femoral midshaft in ovariectomized (OVX) rats were examined. RESULTS: Compared with OVX group, OVX + fluoride-Ca group (O + F) and OVX + Fluoride-Ca + 1,25-dihydroxyvitamin D(O + F + D) two kinds of treatment caused 42%, 91% more in %Tb Ar, 14%, 38% more in Tb Wi, 24%, 46% more in Tb N, 26%, 43% less in Tb Sp, 45%, 42% less in % E Pm, 30%, 60% more in Ob Pm respectively. W Wi in O + F treatment had no significant difference from OVX group but 15.2% in O + F + D treatment compared with sham group, MLT increased 16% in O + F group, had no difference from O + F + D. No significant alteration of mechanical strength at femoral midshaft in both O + F and O + F + D was found. There were significant difference in above changes of parameters except % Ob Pm. CONCLUSIONS: Above two treatments could prevent the loss of cancellous bone, reduced the resorption caused by OVX, protected trabecular microarchitecture, and stimulated bone formation significantly. The combined treatment avoided potential mineralization defect caused by F-Ca treatment alone, and two treatments maintained mechanical strength at femoral midshaft.

Animals↗

PubMed1999

Over-expression of the RAB5 gene in human lung adenocarcinoma cells with high metastatic potential.

The objective of this study is to better understand the molecular mechanism of tumor invasion and metastasis, and isolating tumor metastasis-related genes. Two human lung adenocarcinoma cell lines AGZY-83a and Anip973 were studied. Anip973 was derived from AGZY-83a, but it manifested a very much higher metastatic potential than the parent line. Differential cDNA fragments were isolated by using the techniques of mRNA differential display, and analyzed by means of molecular cloning and sequencing. The expression of RAB5A gene in clinical samples of non-small cell lung cancer was determined by RT-PCR. There were significant differences between AGZY-83a and Anip973 in gene expression. Part of the differential cDNA fragments were cloned and sequenced. We found that there was over-expression of RAB5A gene in the Anip973 cell line. And there was over-expression of RAB5A gene in those samples of clinical lung cancer showing metastasis. In conclusion, the expression or over-expression of RAB5A gene was associated with the metastatic phenotype of Anip973. Probably, over-expression of RAB5A gene in non-small lung cancer may serve as a diagnostic marker for metastasis.

Adenocarcinoma↗

PubMed1998

A novel human gene, WSTF, is deleted in Williams syndrome.

Williams syndrome (WS) is a developmental disorder caused by deletion of multiple genes at chromosome 7q11.23. Here, we report the identification and characterization of a novel gene, WSTF, that maps to the common WS deletion region. WSTF encodes a novel protein of 1425 amino acids with unknown function. It contains one PHD-type zinc finger motif followed by a bromodomain. Both motifs are found in many transcription regulators, suggesting that WSTF may function as a transcription factor. WSTF is ubiquitously expressed in both adult and fetal tissues. The WSTF gene consists of 20 exons spanning about 80 kb. Fluorescence in situ hybridization analysis shows that WSTF is deleted in 50/50 WS individuals. Hemizygous deletion of WSTF may contribute to WS.

Amino Acid Sequence↗

PubMed1998

Osteoblast recruitment and bone formation enhanced by cell matrix-associated heparin-binding growth-associated molecule (HB-GAM).

Bone has an enormous capacity for growth, regeneration, and remodeling. This capacity is largely due to induction of osteoblasts that are recruited to the site of bone formation. The recruitment of osteoblasts has not been fully elucidated, though the immediate environment of the cells is likely to play a role via cell- matrix interactions. We show here that heparin-binding growth-associated molecule (HB-GAM), an extracellular matrix-associated protein that enhances migratory responses in neurons, is prominently expressed in the cell matrices that act as target substrates for bone formation. Intriguingly, N-syndecan, which acts as a receptor for HB-GAM, is expressed by osteoblasts/osteoblast precursors, whose ultrastructural phenotypes suggest active cell motility. The hypothesis that HB-GAM/N-syndecan interaction mediates osteoblast recruitment, as inferred from developmental studies, was tested using osteoblast-type cells that express N-syndecan abundantly. These cells migrate rapidly to HB-GAM in a haptotactic transfilter assay and in a migration assay where HB-GAM patterns were created on culture wells. The mechanism of migration is similar to that previously described for the HB-GAM-induced migratory response of neurons. Our hypothesis that HB-GAM/N-syndecan interaction participates in regulation of osteoblast recruitment was tested using two different in vivo models: an adjuvant-induced arthritic model and a transgenic model. In the adjuvant-induced injury model, the expression of HB-GAM and of N-syndecan is strongly upregulated in the periosteum accompanying the regenerative response of bone. In the transgenic model, the HB-GAM expression is maintained in mesenchymal tissues with the highest expression in the periosteum. The HB-GAM transgenic mice develop a phenotype characterized by an increased bone thickness. HB-GAM may thus play an important role in bone formation, probably by mediating recruitment and attachment of osteoblasts/osteoblast precursors to the appropriate substrates for deposition of new bone.

Animals↗

PubMed1998

Induction of keratinocyte proliferation and lymphocytic infiltration by in vivo introduction of the IL-6 gene into keratinocytes and possibility of keratinocyte gene therapy for inflammatory skin diseases using IL-6 mutant genes.

To understand biological function of IL-6 in the skin in vivo, we constructed a vector that strongly expressed human IL-6 in keratinocytes and introduced it into rat keratinocytes in vivo by the naked DNA method. The overexpression of IL-6 induced macroscopic erythema and histologically evident keratinocyte proliferation and lymphocytic infiltration in the treated area of rat skin. Since previous studies using IL-6 transgenic mice have not shown skin inflammation of these mice, our result provides the first evidence that IL-6 is related to the pathogenesis of inflammatory skin diseases. ELISA suggested that a certain degree of transgenic IL-6 expression in keratinocytes was required for inducing skin inflammation. Cytokine profile in rat keratinocytes after the gene introduction was examined by reverse transcriptase-PCR assay and revealed that gene expression of rat IL-1alpha and TNF-alpha showed no marked change until 24 h, whereas that of rat IL-6 and TGF-alpha increased with time. We then introduced and expressed the IL-6 mutant genes, which were designed to behave as IL-6Ralpha antagonists, and found that their ability to induce erythema was lower than that of the wild-type gene. Furthermore, preintroduction of some mutant genes delayed the erythema induced by postintroduction of the wild-type IL-6 gene, suggesting that the mutant forms of IL-6 prevent wild-type IL-6 from binding to IL-6Ralpha. This result indicates that keratinocyte gene therapy may be possible for inflammatory skin diseases using IL-6 mutant genes.

Animals↗

PubMed1998

Ischemic preconditioning decreases postischemic myocardial tumor necrosis factor-alpha production. Potential ultimate effector mechanism of preconditioning.

BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is an autocrine contributor to myocardial dysfunction and cardiomyocyte death in ischemia-reperfusion (I/R) injury, sepsis, chronic heart failure, and cardiac allograft rejection. Cardiac resident macrophages, infiltrating leukocytes, and cardiomyocytes themselves produce TNF-alpha. Although adenosine reduces macrophage TNF-alpha production and protects myocardium against I/R, it remains unknown whether ischemic preconditioning, which is mediated by adenosine, decreases postischemic myocardial TNF-alpha production. METHODS AND RESULTS: Isolated rat hearts were crystalloid perfused with the Langendorff method and subjected to global, normothermic I/R (20/40 minutes), with or without prior transient ischemic preconditioning (5 minutes) or adenosine pretreatment. Postischemic cardiac TNF-alpha (ELISA) and function were determined (Langendorff). I/R increased cardiac TNF-alpha and impaired myocardial function. Ischemic preconditioning or adenosine decreased myocardial TNF-alpha and improved postischemic functional recovery. Sequestration of myocardial TNF-alpha (TNF binding protein) during the I/R experiments similarly improved postischemic myocardial function. CONCLUSIONS: This study constitutes the initial demonstration that in addition to its other beneficial effects, preconditioning decreases postischemic myocardial TNF-alpha, an autocrine contributor to postischemic myocardial dysfunction. Reduced myocardial TNF-alpha production may represent the distal effector mechanism of preconditioning.

Animals↗