The effect of glucocorticoids on the down-regulation of glucocorticoidreceptor (GR) mRNA was studied in intact rats.GR mRNA was characterized byNorthern blot hybridization and quantitated by dot blot hybridization usi...The effect of glucocorticoids on the down-regulation of glucocorticoidreceptor (GR) mRNA was studied in intact rats.GR mRNA was characterized byNorthern blot hybridization and quantitated by dot blot hybridization using a hu-man GR cDNA fragment as a probe.Administration of hydrocortisone (F) inpolyvinyl alcohol (PVA) resulted in a rapid increase in plasma glucocorticoidswhich maintained at stress levels (20 to 40μg/dl) for about 3 d.HepaticGR mRNA decreased significantly to 73.5±6.3% of control values 6h followingF treatment,after which the decline of GR mRNA was gradual,reaching a mini-mum of 44.0±5.0% of control levels 3d after the treatment.The effect of F onthe down-regulation of hepatic GR mRNA lasted up to 11 d.In contrast,F treat-ment had no effect on GR mRNA in rat brain.These results are consistent withthe changes in GR in rats as reported previously,except that even though thehepatic cytosol GR decreased markedly,no significant changes in hepatic GRmRNA were found 1h after F treatment,strongly suggesting that thedown-regulation of GR by its ligands in vivo occurs at both transcriptional andposttranscriptional levels and is of tissue-specific fashion.展开更多
A simple and rapid method was established to study vascular permeability by in vitroperfused endothelial cell monolayers cultured on micropore filter membrane.It can be used todetermine filtration coefficient (K<s...A simple and rapid method was established to study vascular permeability by in vitroperfused endothelial cell monolayers cultured on micropore filter membrane.It can be used todetermine filtration coefficient (K<sub>f</sub>) to small molecules and osmotic reflection coefficient (σ) toproteins of the endothelial monolayer.Hanks’ balanced salt solution (HBSS) or 5g/L albuminin HBSS was used to perfuse the confluent endothelial monolayer at the hydrostatic pressure of2.45kPa (25cm H<sub>2</sub>O).Control K<sub>f</sub> values were 10.1±0.75 and 3.6±0.75μl·min<sup>-1</sup>·cm<sup>-2</sup>·kPa(-1)(±,n=3) respectively for the perfusion of HBSS and albumin HBSS,suggesting that al-bumin may decrease endothelial monolayer permeability to water and small molecules.After ex-posure of endothelial monolayer to 10<sup>-8</sup>mol/L platelet-activating factor (PAF) for 30min,K<sub>f</sub>values increased to 193.1% and 133.3% respectively.Protein clearance rate (μl.min<sup>-1</sup>·cm<sup>-2</sup>)and osmotic reflection coefficient of the control endothelial monolayer were 8.0±3.22 and 0.37±0.09 respectively and those of the PAF treated endothelial monolayer 12.2±2.95μl·min<sup>-1</sup>·cm<sup>-2</sup> and 0.18±0.06,revealing increased permeability to albumin.Computer-assisted imageprocessing demonstrated that PAF treatment decreased cell area while increased cell form factorand intercellular space,suggesting that endothelial cells retracted and rounded,which may bean important mechanism of PAF-induced increase of vascular permeability.展开更多
We studied the functional changes of pulmonary surfactant (PS) in acutelung injury models produced by endotoxin injection (E.coli O<sub>55</sub>B<sub>5</sub>) in rats.The sur-face properties ...We studied the functional changes of pulmonary surfactant (PS) in acutelung injury models produced by endotoxin injection (E.coli O<sub>55</sub>B<sub>5</sub>) in rats.The sur-face properties of the lung lavage liquid and the total phospholipids (TPL) ex-tracted from it were assessed on a modified Wilhelmy film balance.γ-A isothermof the lavage liquid revealed an increase in minimum surface tension and a de-crease in hysteresis area,recruitment index and stability index,whereas that ofTPL extracted from it did not show any change except for hysteresis area.Thesurface activity correlates positively with the TPL content but negatively with thetotal protein content in the lavage liquid.The findings indicated that there was adysfunction of PS in rats with the lung injury induced by endotoxin,suggestingthat the function deficiency of PS might be caused by decreased phospholipidsand increased proteins in the alveoli.展开更多
The high-affinity glucocorticoid binding sites(HAGS)and the low-affinity glucocor-ticoid binding sites(LAGS)with steroid specificity were demonstrated in cerebral cytosol ofrats by using the radioligand binding as...The high-affinity glucocorticoid binding sites(HAGS)and the low-affinity glucocor-ticoid binding sites(LAGS)with steroid specificity were demonstrated in cerebral cytosol ofrats by using the radioligand binding assay.The equilibrium dissocation constant(Kd)of HAGSand LAGS were(2.78+0.71)×10<sup>-8</sup>mol/L and(2.12±1.06)×10<sup>-6</sup>mol/L respectively as esti-mated by Scatchard and Pseudoseatchard analysis.Glucocorticoid receptors(GR)in the trau-matized(left)hemisphere cytosol were decreased more significantly than those in both the con-trol(right)hemisphere cytosol at 6 h postinjury and normal brain tissue(P【0.05),but Kd ofGR showed no significant changes.GR of liver cytosol at 6h postinjury were more markedly de-creased than normal hepatic cytosol,but Kd of GR underwent no significant changes.These da-ta demonstrate that high-dose glucocorticoid(GC)used in the treatment of traumatic brain ede-ma might maintain target-cell reactions by increasing the production of GC receptor complexesand is most likely to be mediated by LAGS.展开更多
Abstract Objective:To investigate whether hPTH1-34 regulate the synthesis of fibronectin(FN) from cul-trued rat mesangial cells and its possible mechanism.Methods:(1) MCs seeded at a density of 1×10^4 per well in...Abstract Objective:To investigate whether hPTH1-34 regulate the synthesis of fibronectin(FN) from cul-trued rat mesangial cells and its possible mechanism.Methods:(1) MCs seeded at a density of 1×10^4 per well in 24-well plates were treated with medium containing various concentrations of hPTH1-34(10^-12mol/l-10^-8mol/l)for 6h,12h,24h and 48h,control cells were treated with vehicle only.The FN levels (in the supernatant)were measured by ELISA assay.(2) MCs were co-cultured with 10ng/l of anti-TGF-βanti-body and various concentrations of hPTH1-34(10^-12mol/l-10^-6mol/l).Forty-eight hours later, FN were tested by ELISA.(4)MCs were co-cultured with 10ng/l of anti-TGF-β antibody and 10^-8 mol/l hPTH1-34 for 6h,12h,24h and 48h and then FN were tested.Results(1)hPTH1-34 stimulated FN synthesis in a dose-and time-dependent way with a peak at 10^-8 mol/l(P<0.01).(2)Anti-TGF-β antibody inhibited the stimu-lation effect of hPTH1-34 on synthesis of FN in cultured rat mesangial cells(P<0.05).Conclusion:hPTH1-34 up-regulates FN synthesis in cultured rat mesangial cells via TGF-β ,suggesting that PTH may play an im-portant role in deteriorating the residual renal function at the early stage of chroic renal disease.展开更多
The changes of glucocorticoid receptor (GR) and the reactivity of target cells toglucocorticoids (GC) were studied experimentally.The results showed that GC resistance,which wascaused mainly by the decrease of GR,...The changes of glucocorticoid receptor (GR) and the reactivity of target cells toglucocorticoids (GC) were studied experimentally.The results showed that GC resistance,which wascaused mainly by the decrease of GR,developed in burned and shocked rats and dogs.Thesechanges may exacerbate the shock state,and even lead to the development of multiple organfailuue.The protection of GC against intestinal ischemic shock of dogs was demonstrated after theconcentration of dexamethasone (Dex) in plasma was elevated to 10<sup>-6</sup> mol/L by iv injection of 5mgDex/kg body weight.展开更多
The effects of hydrogen peroxide(H<sub>2</sub>O<sub>2</sub>)on endothelial-polymorphonuclear leuko-cyte(EC-PMN)adhesion and their mechanisms were studied in cultured bovine pulmonaryartery ...The effects of hydrogen peroxide(H<sub>2</sub>O<sub>2</sub>)on endothelial-polymorphonuclear leuko-cyte(EC-PMN)adhesion and their mechanisms were studied in cultured bovine pulmonaryartery endothelial monolayers in vitro.H<sub>2</sub>O<sub>2</sub> at various concentrations(10<sup>-3</sup>,10<sup>-2</sup>,10<sup>-1</sup>mol/Lrespectively)stimulated EC-dependent PMN adhesion,of which 10<sup>-2</sup>mol/L H<sub>2</sub>O<sub>2</sub> was the mostpotent one,increasing adhesion to 2.3 times that of the control.Pretreatment of PMNs with SRI63-441,a platelet-activating factor(PAF)receptor antagonist,had no inhibition effect on H<sub>2</sub>O<sub>2</sub>induced EC-PMN adhesion.Pretreatment of ECs with SRI 63-441 before H<sub>2</sub>O<sub>2</sub> exposure signifi-cantly decreased PMN adherence to ECs.Pretreatment of ECs with phospholipase A<sub>2</sub> inhibitorp-bromophenacyl-bromide or calmodulin antagonist chlorpromazine and calcium ion chelate EG-TA obviously decreased H<sub>2</sub>O<sub>2</sub> induced increment of EC-PMN adhesion.These results suggestthat H<sub>2</sub>O<sub>2</sub> may activate ECs,causing the inflow of extracellular calcium or the release of calciumfrom intracellular deposits.Increased intracellar Ca<sup>2+</sup>may bind with calmodulin to activate phos-pholipase A<sub>2</sub>,thus initiating PAF synthesis and promoting EC-PMN adhesion.展开更多
Acute fulminant pancreatitis was produced in dogs by injection of autobile into the mainpancreatic duct.After injection the phospholipase A<sub>2</sub>(PLA<sub>2</sub>)activities in serum,lun...Acute fulminant pancreatitis was produced in dogs by injection of autobile into the mainpancreatic duct.After injection the phospholipase A<sub>2</sub>(PLA<sub>2</sub>)activities in serum,lung lymph andbronchoalveolar lavage fluid(BAL)were elevated significantly,lung lymph flow and pulmonarytransvascular potein clearance increased progressively,protein content and cell numbers in BAL inthe experimental animals were significantly higher than those in the control animals.Furthermore thelung index,wet to dry lung weight ratio,extravascular lung water to bloodless dry lung weight ra-tio,extravascuar lung water to bloodless dry lung weight ratio increased significantly as comparedto control animals.Pretreatment with PLA<sub>2</sub> inhibitor,chloroquine,blocked the changes mentionedabove.This experiment suggests:1.PLA<sub>2</sub> activity in lung lymph fluid as well as in serum andBAL is elevated in acute hemorrhagic pancreatitis.2.Elevated PLA<sub>2</sub> activity may increase thepulmonary vascular permeability.3.PLA<sub>2</sub> is the major factor leading to pulmonary edema in acutehemorrhagic pancreatitis.4.Phagocytes contribute to the lung injury induced by PLA<sub>2</sub> to some ex-tent.展开更多
The homogeneity between the high-and low-affinity glucocorticoid receptors(GR_H and GR_L)was testified by immunoaffinity chromatography using Mab N250(recognizing the immunogenic domain at the N terminal of GR_H)and M...The homogeneity between the high-and low-affinity glucocorticoid receptors(GR_H and GR_L)was testified by immunoaffinity chromatography using Mab N250(recognizing the immunogenic domain at the N terminal of GR_H)and Mab BuGR1(recognizing the DNA binding domain of GR_H).The specific binding peak of 0.98μmol/L[~3H]triamcinolone acetonide(TA)was higher than that of 52.50nmol/L[~3H]TA.The re-sult suggests that the antigenic determinants of GR_L are similar to those of GR_H.ThecDNA of rat liver GR_H was introduced into GR_H-and GR_L-negative mouse fibroblast cellline E82.A3 by calcium phosphate coprecipitation.A number of clones which expressGR_H were selected with G418(400μg/ml).The results of radioligand binding assay indicatethat GR_H gene is expressed successfully and GR_L also may be encoded by GR_H gene.展开更多
Using a model to study vascular permeability under hydrostatically perfused bovine pulmonary artery endothelial cell (EC) monolayers and a software to automatically analyse cell morphological parameters in a computer ...Using a model to study vascular permeability under hydrostatically perfused bovine pulmonary artery endothelial cell (EC) monolayers and a software to automatically analyse cell morphological parameters in a computer image workstation, the effects of isoproterenol (IPN) on platelet-activating factor (PAF)-induced changes in EC monolayer permeability and cell morphological parameters were studied. Albumin has the fortifying effect on endothelial barrier function. After treatment of EC monolayer with 10-8mol/L PAF, trans-monolayer permeability increased, cell surface area decreased, and intercellular space enlarged. As pretreatment with 10-4mol/L IPN, PAF-induced EC permeability increment and morphological changes were blocked. The results suggest that EC contraction and intercellular gap expansion are important mechanisms for PAF-induced high vascular permeability. IPN inhibits the effects of PAF via stabilization of EC morphology and prevention of intercellular gap formation.展开更多
Objective To investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910...Objective To investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910 cells.Methods Activation of the ERK1/2 and p38 was detected by Western blotting using the antibodies against the total ERK1/2 and p38 mitogen-activated protein kinases (MAPKs) protein and the phosphorylated forms of them. Results Dex could suppress the activation of ERK1/2, while enhance the activation of p38 rapidly and strongly in a dose- and time- dependent manner. Neither effect could be blocked by RU486, the antagonist of glucocorticoid receptor (GR).Conclusion Dex has rapid effects on the activation of ERK1/2 and p38, and these effects are not mediated by GR.展开更多
Immune cells, particularly macrophages, play critical roles in the hypoxia-induced inflammatory response. The small GTPase RhoB is usually rapidly induced by a variety of stimuli and has been described as an important...Immune cells, particularly macrophages, play critical roles in the hypoxia-induced inflammatory response. The small GTPase RhoB is usually rapidly induced by a variety of stimuli and has been described as an important regulator of cytoskeletal organization and vesicle and membrane receptor trafficking. However, it is unknown whether RhoB is involved in the hypoxia-induced inflammatory response. Here, we investigated the effect of hypoxia on the expression of RhoB and the mechanism and significance of RhoB expression in macrophages. We found that hypoxia significantly upregulated the expression of RhoB in RAW264.7 cells, mouse peritoneal macrophages, and the spleen of rats. Hypoxia-induced expression of RhoB was significantly blocked by a specific inhibitor of hypoxia-inducible factor-1α (HIF-1α), c-Jun N-terminal kinase (JNK), or extracellular-signal regulated protein kinase (ERK), indicating that hypoxia-activated HIF-1α, JNK, and ERK are involved in the upregulation of RhoB by hypoxia. Knockdown of RhoB expression not only significantly suppressed basal production of interleukin-1 beta (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) in normoxia but also more markedly decreased the hypoxia-stimulated production of these cytokines. Furthermore, we showed that RhoB increased nuclear factor-kappa B (NF-κB) activity, and the inhibition of NF-κB transcriptional activity significantly decreased the RhoB-increased mRNA levels of IL-1β, IL-6, and TNF-α. Finally, we demonstrated that RhoB enhanced cell adhesion and inhibited cell migration in normoxia and hypoxia. Taken together, these results suggest that RhoB plays an important role in the hypoxia-induced activation of macrophages and the inflammatory response.展开更多
Previously,it has been found that glucocorticoid receptor(GR)binding activity decreasedrapidly during heat shock response in HOS-8603,a human osteosarcorna cell line.In this study,Therelationship between the induction...Previously,it has been found that glucocorticoid receptor(GR)binding activity decreasedrapidly during heat shock response in HOS-8603,a human osteosarcorna cell line.In this study,Therelationship between the induction of heat shock proteins(HSPs)and the decrease in GR was furtherstudied in the same cell line.It was found that even though quercetin could specifically inhibit the ex-pression of hsp90α and hsp70 mRNA,it could not prevent GR from the decrease in response to the heatshock treatment.This represents the first reported evidence that the induction of HSPs and the decrease inGR during heat shock response were 2 independent biological events.The results of the present study furthershowed that although the heat shock treatment alone had no effects on alkaline phosphatase(AKP)activity,itcould completely block the induction of AKP activity in HOS-8603 cells by dexamethasone(Dex),a syntheticglucocorticoid.These results demonstrate that the heat shock-induced alteration in GR was accompanied by adecrease in GR functional activity.Furthermore,when the induction of HSPs was inhibited by the treatmentof cells with quercetin,the stimulatory effects of Dex on AKP activity could still be inhibited completely bythe heat shock treatment.The results of this part,on the basis of GR functional activity,further demonstratethat quercetin could not inhibit the heat shock-induced decrease in GR,even though it could inhibit the induc-tion of HSPs.To clarify further the effects of quercetin alone on GR binding activity in HOS-8603 cells,theregulation of GR by quercetin was also studied.It was found for the first time that quercetin coulddown-regulate GR in a time-dependent manner significantly,and that the down-regulation of GR by quercetinin HOS-8603 cells paralelled with a decrease in glucocorticoid-mediated functional responses,suggesting thatthe down-regulation of GR by quercetin is of biological significance.展开更多
The changes of glucocorticoid receptor (GR) during the heat shock response have been studied using a human osteosarcoma cell line (HOS-8603) as the model. The expression of the heat shock protein 70 (hsp70) mRNA in HO...The changes of glucocorticoid receptor (GR) during the heat shock response have been studied using a human osteosarcoma cell line (HOS-8603) as the model. The expression of the heat shock protein 70 (hsp70) mRNA in HOS-8603 cells has been enhanced markedly after a heat treatment at 43 ℃ for 30 min. A mild thermal pretreatment (42℃ for 1 h) protects the HOS-8603 cells against a subsequent heat challenge (46℃). This induced thermotolerance is reflected by the increase of cell viability of HOS-8603 cells. The GR binding activity in HOS-8603 cells decreased rapidly after the heat treatment at 43℃; only 42. 61% of controls were detected 60 min after the heat treatment. However, there was no significant change in the dissociation constant value (Kd). These results indicate that the heat shock induce not only the heat shock mRNA expression, but also the rapid reduction in GR binding activity, suggesting that there might be a functional relationship between GR action and the heat shock response.展开更多
文摘The effect of glucocorticoids on the down-regulation of glucocorticoidreceptor (GR) mRNA was studied in intact rats.GR mRNA was characterized byNorthern blot hybridization and quantitated by dot blot hybridization using a hu-man GR cDNA fragment as a probe.Administration of hydrocortisone (F) inpolyvinyl alcohol (PVA) resulted in a rapid increase in plasma glucocorticoidswhich maintained at stress levels (20 to 40μg/dl) for about 3 d.HepaticGR mRNA decreased significantly to 73.5±6.3% of control values 6h followingF treatment,after which the decline of GR mRNA was gradual,reaching a mini-mum of 44.0±5.0% of control levels 3d after the treatment.The effect of F onthe down-regulation of hepatic GR mRNA lasted up to 11 d.In contrast,F treat-ment had no effect on GR mRNA in rat brain.These results are consistent withthe changes in GR in rats as reported previously,except that even though thehepatic cytosol GR decreased markedly,no significant changes in hepatic GRmRNA were found 1h after F treatment,strongly suggesting that thedown-regulation of GR by its ligands in vivo occurs at both transcriptional andposttranscriptional levels and is of tissue-specific fashion.
文摘A simple and rapid method was established to study vascular permeability by in vitroperfused endothelial cell monolayers cultured on micropore filter membrane.It can be used todetermine filtration coefficient (K<sub>f</sub>) to small molecules and osmotic reflection coefficient (σ) toproteins of the endothelial monolayer.Hanks’ balanced salt solution (HBSS) or 5g/L albuminin HBSS was used to perfuse the confluent endothelial monolayer at the hydrostatic pressure of2.45kPa (25cm H<sub>2</sub>O).Control K<sub>f</sub> values were 10.1±0.75 and 3.6±0.75μl·min<sup>-1</sup>·cm<sup>-2</sup>·kPa(-1)(±,n=3) respectively for the perfusion of HBSS and albumin HBSS,suggesting that al-bumin may decrease endothelial monolayer permeability to water and small molecules.After ex-posure of endothelial monolayer to 10<sup>-8</sup>mol/L platelet-activating factor (PAF) for 30min,K<sub>f</sub>values increased to 193.1% and 133.3% respectively.Protein clearance rate (μl.min<sup>-1</sup>·cm<sup>-2</sup>)and osmotic reflection coefficient of the control endothelial monolayer were 8.0±3.22 and 0.37±0.09 respectively and those of the PAF treated endothelial monolayer 12.2±2.95μl·min<sup>-1</sup>·cm<sup>-2</sup> and 0.18±0.06,revealing increased permeability to albumin.Computer-assisted imageprocessing demonstrated that PAF treatment decreased cell area while increased cell form factorand intercellular space,suggesting that endothelial cells retracted and rounded,which may bean important mechanism of PAF-induced increase of vascular permeability.
文摘We studied the functional changes of pulmonary surfactant (PS) in acutelung injury models produced by endotoxin injection (E.coli O<sub>55</sub>B<sub>5</sub>) in rats.The sur-face properties of the lung lavage liquid and the total phospholipids (TPL) ex-tracted from it were assessed on a modified Wilhelmy film balance.γ-A isothermof the lavage liquid revealed an increase in minimum surface tension and a de-crease in hysteresis area,recruitment index and stability index,whereas that ofTPL extracted from it did not show any change except for hysteresis area.Thesurface activity correlates positively with the TPL content but negatively with thetotal protein content in the lavage liquid.The findings indicated that there was adysfunction of PS in rats with the lung injury induced by endotoxin,suggestingthat the function deficiency of PS might be caused by decreased phospholipidsand increased proteins in the alveoli.
文摘The high-affinity glucocorticoid binding sites(HAGS)and the low-affinity glucocor-ticoid binding sites(LAGS)with steroid specificity were demonstrated in cerebral cytosol ofrats by using the radioligand binding assay.The equilibrium dissocation constant(Kd)of HAGSand LAGS were(2.78+0.71)×10<sup>-8</sup>mol/L and(2.12±1.06)×10<sup>-6</sup>mol/L respectively as esti-mated by Scatchard and Pseudoseatchard analysis.Glucocorticoid receptors(GR)in the trau-matized(left)hemisphere cytosol were decreased more significantly than those in both the con-trol(right)hemisphere cytosol at 6 h postinjury and normal brain tissue(P【0.05),but Kd ofGR showed no significant changes.GR of liver cytosol at 6h postinjury were more markedly de-creased than normal hepatic cytosol,but Kd of GR underwent no significant changes.These da-ta demonstrate that high-dose glucocorticoid(GC)used in the treatment of traumatic brain ede-ma might maintain target-cell reactions by increasing the production of GC receptor complexesand is most likely to be mediated by LAGS.
文摘Abstract Objective:To investigate whether hPTH1-34 regulate the synthesis of fibronectin(FN) from cul-trued rat mesangial cells and its possible mechanism.Methods:(1) MCs seeded at a density of 1×10^4 per well in 24-well plates were treated with medium containing various concentrations of hPTH1-34(10^-12mol/l-10^-8mol/l)for 6h,12h,24h and 48h,control cells were treated with vehicle only.The FN levels (in the supernatant)were measured by ELISA assay.(2) MCs were co-cultured with 10ng/l of anti-TGF-βanti-body and various concentrations of hPTH1-34(10^-12mol/l-10^-6mol/l).Forty-eight hours later, FN were tested by ELISA.(4)MCs were co-cultured with 10ng/l of anti-TGF-β antibody and 10^-8 mol/l hPTH1-34 for 6h,12h,24h and 48h and then FN were tested.Results(1)hPTH1-34 stimulated FN synthesis in a dose-and time-dependent way with a peak at 10^-8 mol/l(P<0.01).(2)Anti-TGF-β antibody inhibited the stimu-lation effect of hPTH1-34 on synthesis of FN in cultured rat mesangial cells(P<0.05).Conclusion:hPTH1-34 up-regulates FN synthesis in cultured rat mesangial cells via TGF-β ,suggesting that PTH may play an im-portant role in deteriorating the residual renal function at the early stage of chroic renal disease.
文摘The changes of glucocorticoid receptor (GR) and the reactivity of target cells toglucocorticoids (GC) were studied experimentally.The results showed that GC resistance,which wascaused mainly by the decrease of GR,developed in burned and shocked rats and dogs.Thesechanges may exacerbate the shock state,and even lead to the development of multiple organfailuue.The protection of GC against intestinal ischemic shock of dogs was demonstrated after theconcentration of dexamethasone (Dex) in plasma was elevated to 10<sup>-6</sup> mol/L by iv injection of 5mgDex/kg body weight.
文摘The effects of hydrogen peroxide(H<sub>2</sub>O<sub>2</sub>)on endothelial-polymorphonuclear leuko-cyte(EC-PMN)adhesion and their mechanisms were studied in cultured bovine pulmonaryartery endothelial monolayers in vitro.H<sub>2</sub>O<sub>2</sub> at various concentrations(10<sup>-3</sup>,10<sup>-2</sup>,10<sup>-1</sup>mol/Lrespectively)stimulated EC-dependent PMN adhesion,of which 10<sup>-2</sup>mol/L H<sub>2</sub>O<sub>2</sub> was the mostpotent one,increasing adhesion to 2.3 times that of the control.Pretreatment of PMNs with SRI63-441,a platelet-activating factor(PAF)receptor antagonist,had no inhibition effect on H<sub>2</sub>O<sub>2</sub>induced EC-PMN adhesion.Pretreatment of ECs with SRI 63-441 before H<sub>2</sub>O<sub>2</sub> exposure signifi-cantly decreased PMN adherence to ECs.Pretreatment of ECs with phospholipase A<sub>2</sub> inhibitorp-bromophenacyl-bromide or calmodulin antagonist chlorpromazine and calcium ion chelate EG-TA obviously decreased H<sub>2</sub>O<sub>2</sub> induced increment of EC-PMN adhesion.These results suggestthat H<sub>2</sub>O<sub>2</sub> may activate ECs,causing the inflow of extracellular calcium or the release of calciumfrom intracellular deposits.Increased intracellar Ca<sup>2+</sup>may bind with calmodulin to activate phos-pholipase A<sub>2</sub>,thus initiating PAF synthesis and promoting EC-PMN adhesion.
文摘Acute fulminant pancreatitis was produced in dogs by injection of autobile into the mainpancreatic duct.After injection the phospholipase A<sub>2</sub>(PLA<sub>2</sub>)activities in serum,lung lymph andbronchoalveolar lavage fluid(BAL)were elevated significantly,lung lymph flow and pulmonarytransvascular potein clearance increased progressively,protein content and cell numbers in BAL inthe experimental animals were significantly higher than those in the control animals.Furthermore thelung index,wet to dry lung weight ratio,extravascular lung water to bloodless dry lung weight ra-tio,extravascuar lung water to bloodless dry lung weight ratio increased significantly as comparedto control animals.Pretreatment with PLA<sub>2</sub> inhibitor,chloroquine,blocked the changes mentionedabove.This experiment suggests:1.PLA<sub>2</sub> activity in lung lymph fluid as well as in serum andBAL is elevated in acute hemorrhagic pancreatitis.2.Elevated PLA<sub>2</sub> activity may increase thepulmonary vascular permeability.3.PLA<sub>2</sub> is the major factor leading to pulmonary edema in acutehemorrhagic pancreatitis.4.Phagocytes contribute to the lung injury induced by PLA<sub>2</sub> to some ex-tent.
文摘The homogeneity between the high-and low-affinity glucocorticoid receptors(GR_H and GR_L)was testified by immunoaffinity chromatography using Mab N250(recognizing the immunogenic domain at the N terminal of GR_H)and Mab BuGR1(recognizing the DNA binding domain of GR_H).The specific binding peak of 0.98μmol/L[~3H]triamcinolone acetonide(TA)was higher than that of 52.50nmol/L[~3H]TA.The re-sult suggests that the antigenic determinants of GR_L are similar to those of GR_H.ThecDNA of rat liver GR_H was introduced into GR_H-and GR_L-negative mouse fibroblast cellline E82.A3 by calcium phosphate coprecipitation.A number of clones which expressGR_H were selected with G418(400μg/ml).The results of radioligand binding assay indicatethat GR_H gene is expressed successfully and GR_L also may be encoded by GR_H gene.
基金Project supported by the National Natural Science Foundation of China.
文摘Using a model to study vascular permeability under hydrostatically perfused bovine pulmonary artery endothelial cell (EC) monolayers and a software to automatically analyse cell morphological parameters in a computer image workstation, the effects of isoproterenol (IPN) on platelet-activating factor (PAF)-induced changes in EC monolayer permeability and cell morphological parameters were studied. Albumin has the fortifying effect on endothelial barrier function. After treatment of EC monolayer with 10-8mol/L PAF, trans-monolayer permeability increased, cell surface area decreased, and intercellular space enlarged. As pretreatment with 10-4mol/L IPN, PAF-induced EC permeability increment and morphological changes were blocked. The results suggest that EC contraction and intercellular gap expansion are important mechanisms for PAF-induced high vascular permeability. IPN inhibits the effects of PAF via stabilization of EC morphology and prevention of intercellular gap formation.
文摘Objective To investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910 cells.Methods Activation of the ERK1/2 and p38 was detected by Western blotting using the antibodies against the total ERK1/2 and p38 mitogen-activated protein kinases (MAPKs) protein and the phosphorylated forms of them. Results Dex could suppress the activation of ERK1/2, while enhance the activation of p38 rapidly and strongly in a dose- and time- dependent manner. Neither effect could be blocked by RU486, the antagonist of glucocorticoid receptor (GR).Conclusion Dex has rapid effects on the activation of ERK1/2 and p38, and these effects are not mediated by GR.
文摘Immune cells, particularly macrophages, play critical roles in the hypoxia-induced inflammatory response. The small GTPase RhoB is usually rapidly induced by a variety of stimuli and has been described as an important regulator of cytoskeletal organization and vesicle and membrane receptor trafficking. However, it is unknown whether RhoB is involved in the hypoxia-induced inflammatory response. Here, we investigated the effect of hypoxia on the expression of RhoB and the mechanism and significance of RhoB expression in macrophages. We found that hypoxia significantly upregulated the expression of RhoB in RAW264.7 cells, mouse peritoneal macrophages, and the spleen of rats. Hypoxia-induced expression of RhoB was significantly blocked by a specific inhibitor of hypoxia-inducible factor-1α (HIF-1α), c-Jun N-terminal kinase (JNK), or extracellular-signal regulated protein kinase (ERK), indicating that hypoxia-activated HIF-1α, JNK, and ERK are involved in the upregulation of RhoB by hypoxia. Knockdown of RhoB expression not only significantly suppressed basal production of interleukin-1 beta (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) in normoxia but also more markedly decreased the hypoxia-stimulated production of these cytokines. Furthermore, we showed that RhoB increased nuclear factor-kappa B (NF-κB) activity, and the inhibition of NF-κB transcriptional activity significantly decreased the RhoB-increased mRNA levels of IL-1β, IL-6, and TNF-α. Finally, we demonstrated that RhoB enhanced cell adhesion and inhibited cell migration in normoxia and hypoxia. Taken together, these results suggest that RhoB plays an important role in the hypoxia-induced activation of macrophages and the inflammatory response.
文摘Previously,it has been found that glucocorticoid receptor(GR)binding activity decreasedrapidly during heat shock response in HOS-8603,a human osteosarcorna cell line.In this study,Therelationship between the induction of heat shock proteins(HSPs)and the decrease in GR was furtherstudied in the same cell line.It was found that even though quercetin could specifically inhibit the ex-pression of hsp90α and hsp70 mRNA,it could not prevent GR from the decrease in response to the heatshock treatment.This represents the first reported evidence that the induction of HSPs and the decrease inGR during heat shock response were 2 independent biological events.The results of the present study furthershowed that although the heat shock treatment alone had no effects on alkaline phosphatase(AKP)activity,itcould completely block the induction of AKP activity in HOS-8603 cells by dexamethasone(Dex),a syntheticglucocorticoid.These results demonstrate that the heat shock-induced alteration in GR was accompanied by adecrease in GR functional activity.Furthermore,when the induction of HSPs was inhibited by the treatmentof cells with quercetin,the stimulatory effects of Dex on AKP activity could still be inhibited completely bythe heat shock treatment.The results of this part,on the basis of GR functional activity,further demonstratethat quercetin could not inhibit the heat shock-induced decrease in GR,even though it could inhibit the induc-tion of HSPs.To clarify further the effects of quercetin alone on GR binding activity in HOS-8603 cells,theregulation of GR by quercetin was also studied.It was found for the first time that quercetin coulddown-regulate GR in a time-dependent manner significantly,and that the down-regulation of GR by quercetinin HOS-8603 cells paralelled with a decrease in glucocorticoid-mediated functional responses,suggesting thatthe down-regulation of GR by quercetin is of biological significance.
文摘The changes of glucocorticoid receptor (GR) during the heat shock response have been studied using a human osteosarcoma cell line (HOS-8603) as the model. The expression of the heat shock protein 70 (hsp70) mRNA in HOS-8603 cells has been enhanced markedly after a heat treatment at 43 ℃ for 30 min. A mild thermal pretreatment (42℃ for 1 h) protects the HOS-8603 cells against a subsequent heat challenge (46℃). This induced thermotolerance is reflected by the increase of cell viability of HOS-8603 cells. The GR binding activity in HOS-8603 cells decreased rapidly after the heat treatment at 43℃; only 42. 61% of controls were detected 60 min after the heat treatment. However, there was no significant change in the dissociation constant value (Kd). These results indicate that the heat shock induce not only the heat shock mRNA expression, but also the rapid reduction in GR binding activity, suggesting that there might be a functional relationship between GR action and the heat shock response.