Biomedical subjects

C J Jen

Publications and source records attributed to C J Jen.

At least 19 recordsLinked to original sources

PubMed2005

Compulsive exercise acutely upregulates rat hippocampal brain-derived neurotrophic factor.

This study was to examine the effects of treadmill exercise on the expression of brain-derived neurotrophic factor (BDNF) in rat hippocampus. After 1-wk treadmill familiarization, animals in exercise groups received a 4-wk exercise training or an acute exercise. They were sacrificed 2 h or 2 d after exercise and their hippocampal BDNF mRNA and protein levels were determined. We demonstrated that 1) hippocampal BDNF mRNA and protein levels were both elevated in response to exercise training at 2 h after the last run but not after 2 d; 2) an acute moderate exercise (1 or 3 d) increased BDNF protein levels; 3) acute severe exercise increased BDNF protein and mRNA levels in animals under a familiarization regimen, while suppressed the BDNF mRNA level in rats without treadmill familiarization, paralleling the stress effect of immobilization/water exposure. We conclude that compulsive treadmill exercise with pre-familiarization acutely upregulates rat hippocampal BDNF gene expression.

Animals↗

PubMed2001

C57BL/6 and BALB/c bronchoalveolar macrophages respond differently to exercise.

Macrophages from prototypical Th1 strains (e.g., C57BL/6) and Th2 strains (e.g., BALB/c) are classified as M-1 and M-2 phenotypes. We investigated the different phagocytic responses between M-1 and M-2 bronchoalveolar macrophages (BAMs) under resting and two various exercise conditions. At rest, M-1 BAMs showed higher phagocytic capacity of unopsonized particles, higher expression of MARCO (macrophage receptor with collagenous structure), and higher generation of NO than M-2 BAMs. Severe exercise, but not moderate exercise, significantly enhanced both phagocytosis of unopsonized particles and expression of MARCO in M-2 BAMs. In contrast, M-1 BAMs were unaffected by either exercise protocol. The phagocytosis of unopsonized particles was largely mediated by MARCO, especially in M-1 BAMs. Secreted products from cultured M-2 BAMs isolated after severe exercise, but not those from M-1 BAMs, enhanced BAM phagocytosis. The cultured M-1 BAMs secreted phagocytosis inhibitors, and this effect could be blocked by NO antagonists. Moreover, the extent of phagocytosis suppression induced by M-1 BAM-secreted products correlated with their production of nitrite/nitrate. Exogenous NO donors as well as NO derivatives, nitrite and nitrate, suppressed the BAM phagocytosis. We propose that while the severe exercise-enhanced phagocytosis in M-2 BAMs was largely mediated by MARCO up-regulation and secretion of stimulators, the lack of exercise effect in M-1 BAMs could be partially due to the constitutive secretion of NO-related suppressors. In conclusion, genetically different mice use different strategies in regulating BAM activity under resting conditions and in response to various exercise paradigms.

Animals↗

PubMed2001

Endothelial [Ca2+]i is an integrating signal for the vascular tone in rat aortae.

BACKGROUND: Although various endothelium-dependent relaxing factors (endothelial autacoids) are released upon the elevation of endothelial cytosolic free Ca2+ concentration (EC [Ca2+]i), the quantitative relationship between EC [Ca2+]i and vascular tone remains to be established. Moreover, whether the basal release of endothelial autacoids is modulated by basal EC [Ca2+]i is still unclear. We assessed these issues by using a novel method that allows simultaneous recording of EC [Ca2+]i and vascular displacement in dissected rat aortic segments. RESULTS: Receptor-dependent (acetylcholine) or independent (ionomycin) agonists caused immediate EC [Ca2+]i elevation followed by vasorelaxation in preparations pre-contracted with phenylephrine. Low doses of agonists induced small EC [Ca2+]i elevations (about 100 nmol/L) and concomitant half-maximal vasorelaxation. At high doses, agonists elevated EC [Ca2+]i to micromol/L range with little additional vasodilatation. When EC [Ca2+]i was plotted against the vasorelaxation, the curves were almost identical for both acetylcholine and ionomycin treatments, in the presence or absence of various endothelial autacoid inhibitors. Calcium-free solution reduced basal EC [Ca2+]i and induced a drastic vasoconstriction. Endothelial autacoid inhibitors reduced EC [Ca2+]i changes and abolished both agonist-induced vasodilatation and calcium-free solution-induced vessel contraction. When the EC [Ca2+]i was completely chelated by 40 micromol/L BAPTA, the acetylcholine-evoked vasorelaxation could be abolished as well. However, when the EC [Ca2+]i was partially chelated by 20 micromol/L BAPTA, the acetylcholine-evoked vasorelaxation was almost unaffected. CONCLUSIONS: These results indicate that vascular tone is modulated by subtle changes of EC [Ca2+]i level, which seems to serve as an integrating signal in both basal and stimulated states.

Animals↗

PubMed2001

Expression in Pichia pastoris and characterization by circular dichroism and NMR of rhodostomin.

Rhodostomin (Rho) is a snake venom protein isolated from Calloselasma rhodostoma. Rho is a disintegrin that inhibits platelet aggregation by blocking the binding of fibrinogen to the integrin alpha(IIb)beta3 of platelets. Rho produced in Escherichia coli inhibited platelet aggregation with a K(I) value of 263 nM. Although functional, Rho produced in E. coli is misfolded based on our 2D and 3D NMR studies. In order to correct the folding problem, Rho was expressed in Pichia pastoris. The recombinant Rho expressed in P. pastoris inhibited platelet aggregation with a resulting K(I) value of 70 nM. This is the same potency as that of native Rho. CD analysis showed that the secondary structures of Rho are pH-independent and contain 3.5-7.9% alpha-helix, 48.2-50.5% beta-structures, and 42.3-47% coil. The sequential assignment and structure analysis of Rho were obtained using 2D and 3D 15N-edited NMR spectra. These results provide the first direct evidence that highly disulfide-bonded disintegrin can be expressed in P. pastoris with the correct fold. This evidence may serve as the basis for exploring the structure and function relationships as well as the dynamics of disintegrin and its variants.

Amino Acid Sequence↗

PubMed2001

Severe exercise enhances phagocytosis by murine bronchoalveolar macrophages.

Because physical activity affects the immune competency of individuals by an unknown mechanism, we investigated the effect of acute exercise on phagocytosis of bronchoalveolar macrophages (BAMs). Male BALB/c mice, 7-9 weeks old, ran on a treadmill to exhaustion (severe exercise, SE) or at a final speed of 17 m/min for 30 min (moderate exercise, ME). Although both exercise protocols induced differential leukocytosis, 95% leukocytes from lung lavages of both groups were BAMs. The BAM phagocytic capacity of nonopsonized beads increased immediately after SE but not after ME, gradually returning to the basal level after 4 h. SE upregulates the macrophage scavenger receptors (SR-A type I/II and MARCO), CR3, and ICAM-1, but not Fc gammaR. Although the blocking effect of MARCO antibody was most pronounced, that of ICAM-1 antibody was totally reversed by cross-linking CR3. Our results showed that SE, but not ME, activated BAMs and that the enhanced nonopsonized phagocytosis was mainly mediated by scavenger receptors and ICAM-1/CR3.

Animals↗

PubMed2000

Endothelial [Ca(2+)](i) signaling during transmigration of polymorphonuclear leukocytes.

Vascular endothelium plays an important role in regulating the transendothelial migration of polymorphonuclear leukocytes (PMNs). In this study, the intracellular calcium ion ([Ca(2+)](i)) signaling of endothelial cells (ECs) during PMN transmigration was examined at the single-cell level. Human umbilical vein ECs were cultured on a thin layer of collagen gel. The ECs were labeled with fura-2, immersed in formyl-Met-Leu-Phe, and subsequently perfused with fresh buffer to establish a gradient of chemoattractant across the EC monolayer. The entire process of PMN rolling on, adhering to, and transmigrating across the EC monolayer was recorded under both phase-contrast and fluorescence optics. The data showed the following: (1) At high concentration (approximately 3 x 10(6)/mL), both PMN suspension and its supernatant stimulated frequent EC [Ca(2+)](i) elevations across the monolayer; (2) when used at lower concentration (approximately 5 x 10(5)/mL) to avoid the interference of soluble factors, PMN transmigration, but not rolling or adhesion, was accompanied by EC [Ca(2+)](i) elevation; (3) the latter EC [Ca(2+)](i) elevation occurred simultaneously in ECs adjacent to the transmigration site, but not in those that were not in direct contact with the transmigrating PMNs; (4) this EC [Ca(2+)](i) elevation was an initial and required event for PMN transmigration; and (5) PMNs pretreated with 5,5'-dimethyl-1, 2-bis(2-aminophenoxy)ethane-N, N, N', N'-tetraacetic acid transmigrated with the accompanying EC [Ca(2+)](i) elevation, but they became elongated in the collagen gel. In conclusion, PMNs induce adjacent EC [Ca(2+)](i) signaling, which apparently mediates the "gating" step for their subsequent transmigration. (Blood. 2000;96:3816-3822)

Calcium↗

PubMed2000

Effects of Welsh onion extracts on human platelet function in vitro.

Welsh onion has been consumed for prevention of cardiovascular disorders. However, its underlying mechanisms are still unclear. This study investigated whether Welsh onion extracts can alter human platelet function (ie, platelet adhesion, aggregation, and thromboxane release). To clarify the underlying mechanisms, we also measured the intracellular calcium ([Ca2+]i) and cyclic nucleotide levels in platelets. Our results showed that 1) boiled extracts directly induced platelet aggregation in a dose-dependent manner; 2) raw extracts inhibited platelet adhesion and ADP-evoked platelet aggregation, while boiled extracts enhanced them; 3) raw green extract suppressed ADP-stimulated platelet [Ca2+]i elevation and thromboxane production, whereas boiled green extract enhanced them; 4) raw green extract elevated platelet cAMP level, whereas boiled green extract had no effect on cAMP level. Furthermore, the boiled green extract, but not the raw extract, induced pronounced platelet morphological changes. In conclusion, raw extracts of Welsh onion inhibit platelet function in vitro while boiled extracts activate platelets.

Blood Platelets↗

PubMed2000

Chronic consumption of raw but not boiled Welsh onion juice inhibits rat platelet function.

Welsh onion has been consumed for prevention of cardiovascular disorders. To study if it has antithrombotic effects, 9-wk-old male Sprague-Dawley rats were studied. Some rats were fed raw or boiled Welsh onion juice (2 g. kg(-1). d(-1)) for 4 wk, and the remaining acted as the control. Before and after feeding, their systolic blood pressure was measured by a tail-cuff method. Two days after the treatment period, tail bleeding time, platelet function (including platelet aggregation and adhesion), plasma levels of prostaglandins, and platelet cyclic nucleotide levels were determined. In comparison to the control, raw Welsh onion juice consumption significantly (1) lowered resting systolic blood pressure; (2) prolonged the bleeding time; (3) diminished platelet adhesion on a fibrinogen-coated surface, ADP-evoked platelet aggregation and ADP-stimulated thromboxane release; (4) elevated the concentration of cyclic AMP, but not cyclic GMP, in platelets; (5) increased the plasma level of 6-keto-prostaglandin F(1alpha), the stable prostacyclin metabolite, but not the plasma nitrite level. On the contrary, boiled Welsh onion juice consumption was totally ineffective. In conclusion, consuming raw Welsh onion juice, but not boiled juice, has blood pressure lowering and antithrombotic effects in rats. These effects may be mediated by PGI(2)-cAMP pathway.

6-Ketoprostaglandin F1 alpha↗

PubMed2000

Heterogeneity of [Ca(2+)](i) signaling in intact rat aortic endothelium.

Most existing knowledge about [Ca(2+)](i) signaling in vascular endothelium has been based on studies using endothelial cells cultured in vitro. To examine how endothelial cells behave in situ, we have developed a method to monitor single-cell [Ca(2+)](i) from Fura-2-loaded rat aortic segments. Fluorescence ratio images from large numbers of endothelial cells were acquired by using a flow chamber mounted on a dual-wavelength fluorescence microscope. Our results showed that either acetylcholine or histamine reversibly activated the vascular endothelium by eliciting M(3) or H(1) receptor-mediated [Ca(2+)](i) increases, respectively. The acetylcholine-evoked endothelial [Ca(2+)](i) elevation at the branch site (intercostal orifice) was much more pronounced than that at the non-branch area. However, endothelium at the branch site was relatively insensitive to histamine. Both acetylcholine-sensitive and histamine-sensitive endothelial cells were arranged in belts aligned along flow lines and were intercalated with each other. Data analyzed from 400 endothelial cells located at the non-branch site showed drastically heterogeneous [Ca(2+)](i) responses to a fixed concentration of either acetylcholine or histamine, differing by two orders of magnitude in individual cells. As a conclusion, vascular endothelial cells appear to have their own characteristic [Ca(2+)](i) 'fingerprint' to various agonists and they may function coordinately in situ.

Acetylcholine↗

PubMed2000

Effects of chronic exercise on calcium signaling in rat vascular endothelium.

Chronic exercise enhances endothelium-dependent vasodilating responses. To investigate whether this is due to a change in endothelial Ca(2+) signaling, we examined intracellular Ca(2+) concentration ([Ca(2+)](i)) level in rat aortic endothelium in response to acetylcholine (ACh) or ATP. Four-week-old male Wistar rats were divided into control and exercise groups. The exercised animals ran on a treadmill at a moderate intensity for 60 min/day, 5 day/wk, for 10 wk. Rat aortas were then excised and loaded with fura 2. After the aortas were mounted on a flow chamber, these specimens were observed under an epifluorescence microscope equipped with ratio-imaging capability. Our results showed that 1) chronic exercise increased both ACh- and ATP-induced [Ca(2+)](i) responses; 2) ACh induced heterogeneous [Ca(2+)](i) elevation in individual endothelial cells; and 3) the exercise effect on ACh-evoked endothelial [Ca(2+)](i) elevation was inhibited by the Ca(2+) influx blocker SKF-96365, by a Ca(2+)-free buffer, or by high concentrations of extracellular K(+). We conclude that chronic exercise increases ACh-induced [Ca(2+)](i) elevation in rat aortic endothelium in situ, possibly by facilitating Ca(2+) influx.

Acetylcholine↗

PubMed2000

Invited review: effects of flow on vascular endothelial intracellular calcium signaling of rat aortas ex vivo.

To study the effects of flow on in situ endothelial intracellular calcium concentration ([Ca(2+)](i)) signaling, rat aortic rings were loaded with fura 2, mounted on a tissue flow chamber, and divided into control and flow-pretreated groups. The latter was perfused with buffer at a shear stress of 50 dyns/cm(2) for 1 h. Endothelial [Ca(2+)](i) responses to ACh or shear stresses were determined by ratio image analysis. Moreover, ACh-induced [Ca(2+)](i) elevation responses were measured in a calcium-free buffer, or in the presence of SKF-96365, to elucidate the role of calcium influx in the flow effects. Our results showed that 1) ACh increased endothelial [Ca(2+)](i) in a dose-dependent manner, and these responses were incremented by flow-pretreatment; 2) the differences in ACh-induced [Ca(2+)](i) elevation between control and flow-pretreated groups were abolished by SKF-96365 or by Ca(2+)-free buffer; and 3) in the presence of 10(-5) M ATP, shear stress induced dose-dependent [Ca(2+)](i) elevation responses that were not altered by flow-pretreatment. In conclusion, flow-pretreatment augments the ACh-induced endothelial calcium influx in rat aortas ex vivo.

Acetylcholine↗

PubMed1999

Chronic exercise enhances vascular responses to clonidine in rats by increasing endothelial alpha2-adrenergic receptor affinity.

Chronic exercise increases endothelium-dependent vasodilating responses. To investigate whether endothelial alpha2-adrenergic receptor upregulation is involved in the enhancement of clonidine-induced vasorelaxation by chronic exercise, 4-week-old male Wistar rats were used. They were divided into control and exercise groups. The trained animals ran on a treadmill at a moderate intensity for 60 min per day, 5 days per week for 10 weeks in total. Resting heart rates were measured by a tail-cuff method to confirm training effects. After training, rings of the thoracic aorta were prepared to evaluate vasodilating responses to clonidine, an alpha2 agonist. Released endothelium-derived relaxing factors were pharmacologically identified by treatment of N(omega)-nitro-L-arginine, a nitric oxide (NO) synthase inhibitor, or tetraethylammonium chloride, an endothelium-derived hyperpolarization factor (EDHF) inhibitor. Receptor binding assays were performed by using 3H-labeled clonidine as a tracer. We found that chronic exercise enhanced vascular responses to clonidine by stimulating the release of both NO and EDHF. It also increased the binding affinity of endothelial cell alpha2 receptor without changing the number of binding sites. Therefore, the elevated vasorelaxing responses to clonidine after chronic exercise may be partially resulted from an increase in endothelial alpha2 receptor binding affinity.

Adrenergic alpha-Agonists↗

PubMed1999

Acute exercise enhances receptor-mediated endothelium-dependent vasodilation by receptor upregulation.

The effects of acute exercise on receptor-mediated endothelium-dependent vasodilation and its possible mechanisms were investigated in the presence of indomethacin. Male Wistar rats (16-20 weeks old) were divided into control and exercise groups. The exercise group ran on a drum exerciser until exhaustion, followed by immediate decapitation. Acetylcholine (ACh)- or clonidine (CLO)-induced vasodilating responses in thoracic aortae of the control and exercise groups were compared. Receptor-binding assays were performed to determine whether there were any upregulations of endothelial receptors after acute exercise. Our results indicated that acute exercise induced the following effects: (1) the dose-response curves of ACh and CLO shifted to the left; (2) the high-affinity M3 binding sites increased in number but not in affinity; (3) the alpha2 binding sites decreased in number but increased in affinity. We conclude that acute exercise enhances receptor-mediated vasodilation responses, at least in part, by regulating either endothelial receptor number or receptor affinity.

Acetylcholine↗

PubMed1997

Effects of age and hypertension on endothelium-dependent vasodilating responses.

To investigate whether the release of endothelium-derived relaxing factors (EDRF) was affected during the development of hypertension or by age, male spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) in the age of 4, 8, 12, 24, 36 or 48 weeks old were used for this study. The thoracic aorta and superior mesenteric arteries of these animals with different ages were excised after general anesthesia. In addition to the measurement of basal EDRF release, vascular responses to acetylcholine (ACh) were assessed with or without the treatment of various inhibitors (such as N omega-nitro-L-arginine, SQ29548 or 3-amino-1,2,4-triazole) to clarify the possible mechanisms for changes of ACh-evoked vasorelaxation. We found that 1) the basal release of EDRF was declined during the development of hypertension, especially in the mesenteric arteries; 2) ACh-induced vasorelaxation in the thoracic aorta was mainly due to the stimulated release of nitric oxide, whereas the effect of endothelium-derived hyperpolarizing factor was more prominent in the mesenteric arteries than in thoracic aortae; 3) high concentrations of ACh stimulated the release of endothelium-derived contracting factors in the thoracic aorta of SHR and WKY of 24 weeks or older, and in the mesenteric arteries of 48-week-old SHR. In conclusion, basal release of EDRF decreases before hypertension is well established, and the impairment of ACh-evoked vasorelaxation in adult SHR is mainly due to the release of contracting factors.

Acetylcholine↗

PubMed1997

Effects of chronic exercise and deconditioning on platelet function in women.

To investigate the effects of chronic exercise and deconditioning on platelet function in women, 16 healthy sedentary women were divided into control and exercise groups. The exercise group cycled on an ergometer at 50% maximal oxygen consumption for 30 min/day, 5 days/wk, for two consecutive menstrual cycles and then were deconditioned for three menstrual cycles. During this period, platelet adhesiveness on a fibrinogen-coated surface, ADP-induced platelet aggregation and intracellular calcium concentration elevation, guanosine 3',5'-cyclic monophosphate (cGMP) content in platelets, and plasma nitric oxide metabolite levels were measured before and immediately after a progressive exercise test in the midfollicular phase. Our results indicated that, after exercise training, 1) resting heart rates and blood pressures were reduced, and exercise performance was improved; 2) resting platelet function was decreased, whereas plasma nitrite and nitrate levels and platelet cGMP contents were enhanced; and 3) the potentiation of platelet function by acute strenuous exercise was decreased, whereas the increases in plasma nitrite and nitrate levels and platelet cGMP contents were enhanced by acute exercise. Furthermore, deconditioning reversed these training effects. This implies that training-induced platelet functional changes in women in the midfollicular phase may be mediated by nitric oxide.

Adult↗

PubMed1997

Effects of short-term exercise on female platelet function during different phases of the menstrual cycle.

Previous studies have shown that premenopausal women have a low incidence of cardiovascular diseases, and that acute exercise affects male platelet function in an intensity-dependent manner. To investigate whether acute exercise affects female platelet function differently from males, sixteen sedentary women in the midfollicular phase or midluteal phase received strenuous or moderate exercise on a bicycle ergometer. Before and immediately after exercise, platelet adhesiveness, adenosine diphosphate-induced platelet aggregation and intracellular calcium concentration elevation, platelet cAMP and cGMP contents, urinary 11-dehydro-TXB2 and 6-keto-prostaglandin F1 alpha levels, and plasma nitric oxide metabolite level were determined. Our results showed no differences in exercise performance and in resting platelet function between two menstrual phases, with little change in urinary eicosanoid metabolites and platelet cAMP levels under all experimental conditions. In addition, for women in the midfollicular phase, (1) strenuous exercise increased platelet adhesiveness, adenosine-diphosphate-induced platelet aggregation, and intracellular calcium concentration elevation, whereas moderate exercise suppressed them; (2) moderate exercise enhanced plasma nitric oxide metabolite and platelet cGMP levels. In contrast, none of these platelet functions was affected by acute exercise in the midluteal phase. Therefore, we conclude that acute exercise affects female platelet function in an intensity-dependent manner in the midfollicular phase but not in the midluteal phase. The irresponsiveness of platelets to acute exercise in the luteal phase may partially explain why premenopausal women have a lower incidence of cardiovascular diseases than men.

Adult↗

PubMed1997

Perturbation of platelet adhesion to endothelial cells by plasminogen activation in vitro.

To investigate whether the endothelium-platelet interactions may be altered by plasminogen activation, cultured human umbilical vein endothelial cells (ECs) were treated with tissue-type plasminogen activator (t-PA) in the presence of plasminogen, and platelet adhesion to ECs was subsequently measured by using a tapered flow chamber. Our results demonstrated that platelets adhered more readily to t-PA treated EC monolayer than to the control monolayer at all shear stress levels tested. This phenomenon was treatment time-dependent and dose-dependent, and it could be blocked by adding plasmin inhibitors, such as epsilon-amino caproic acid and aprotinin. Adherent platelets on t-PA treated EC monolayer underwent more severe shape change than those on the control monolayer. While the extracellular matrix directly treated with t-PA attracted less platelets than the control matrix did, platelet adhesion to the matrix that was produced by t-PA-treated ECs was unaltered. These data suggest that t-PA treatment on ECs compromised antiplatelet-adhesion capability on their apical surface without altering the reactivity of their extracellular matrix towards platelets.

Blood Platelets↗

PubMed1996

Changes in cytosolic calcium concentrations and cell morphology in single platelets adhered to fibrinogen-coated surface under flow.

Changes in intracellular calcium concentration [Ca2+]i of fura-2-loaded human platelet during its adhesion to a fibrinogen-coated surface were studied, using a flow chamber mounted on an epifluorescence microscope equipped with digital-ratio imaging. Adherent platelets were individually mapped under a scanning electron microscope to establish the possible correlation between adhesion-associated shape alterations and [Ca2+]i changes. We found that 1) there was no immediate [Ca2+]i elevation on platelet adhesion; 2) [Ca2+]i changes varied drastically platelets with a lag time ranging 10 to 200 s, averaging about 1 minute; 3) the pattern of [Ca2+]i changes varied drastically among individual adherent platelets; 4) the degree of [Ca2+]i elevation appeared to correlate with the extent of morphology change, with the vast majority ( > 90%) of spread platelets showed detectable [Ca2+]i changes; 5) neither morphological nor [Ca2+]i changes correlated with the lag time; 6) platelets treated with dimethyl-BAPTA (15 mumol/L) underwent normal shape change without [Ca2+]i elevation; 7) cytochalasin D (10 mumol/L) inhibited both shape change and [Ca2+]i elevation; 8) colchicine (1 mmol/L) was ineffective in both regards. We conclude that although platelet adhesion-associated shape changes may be accompanied with heterogeneous [Ca2+]i changes that are microfilament-dependent, [Ca2+]i changes do not happen immediately after platelet-surface contact and they are not required for adherent platelets to undergo postcontact morphological changes.

Blood Platelets↗