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Biomedical subjects

E B Chang

Publications and source records attributed to E B Chang.

At least 73 records · Page 4Linked to original sources
PubMed1991

Cystic fibrosis transmembrane regulator mRNA expression relative to ion-nutrient transport in spontaneously differentiating human intestinal CaCo-2 epithelial cells.

The relative abundance of cystic fibrosis transmembrane regulator (CFTR) message at various stages of postconfluence development was compared with evolving cellular ion-nutrient transport properties of CaCo-2 human intestinal cells. Initially these cells demonstrate electrogenic Cl secretion manifested by secretagogue-induced changes in short-circuit current. Over time, however, the secretory characteristics of CaCo-2 monolayers diminish, and brush border hydrolase activities and glucose-dependent and amiloride-sensitive Na transport increase. With a polymerase chain reaction-derived cDNA probe to CFTR exon 13, two distinct mRNA transcripts of 6.5 and 4.3 kb were found. No significant differences in their abundance were noted in cells at 6 and 28 days after confluence. These data suggest two possible interpretations for the role of CFTR protein. First, if CFTR is membrane Cl channel, its protein expression or activity could be differentially regulated during CaCo-2 cell development. Alternatively, CFTR may not be a Cl channel but may serve as an important regulatory membrane protein.

Amiloride↗
PubMed1990

Phorbol ester-stimulated secretion in chicken ileum: role of arachidonic acid metabolism.

Phorbol 12,13-dibutyrate, an activator of protein kinase C, stimulates electrogenic anion secretion in chicken ileum. This effect can be inhibited by the cyclooxygenase inhibitor piroxicam, suggesting arachidonic acid metabolites as mediators. Phorbol 12,13-dibutyrate stimulates increases in prostaglandin E2 release and mucosal 3',5'-cyclic adenosine monophosphate content in intact mucosa. Stimulation of prostaglandin E2 release appears to come mainly from the subepithelial compartment and not from epithelial cells. Unlike its effect on anion secretion, phorbol 12,13-dibutyrate inhibition of sodium uptake by sodium-hydrogen exchange in isolated mature villus enterocytes is not inhibited by piroxicam. This study concludes that phorbol ester effects on anion secretion in chickens appear to be mediated by arachidonic acid metabolites most likely produced by cells of the lamina propria and submucosa, whereas phorbol ester effects on inhibiting upper villus cell sodium-hydrogen exchange may be a direct effect of phorbol esters.

Animals↗
PubMed1990

Interleukins 1 and 3 stimulate anion secretion in chicken intestine.

Interleukin 1 or 3 added to the serosal side of chicken small intestine transiently increases short-circuit current. Replacement of bathing-medium Cl and HCO3 with gluconate and HEPES abolished the short-circuit current increase, consistent with these cytokines stimulating electrogenic anion secretion. Cytokine-stimulated short-circuit current changes were inhibited by preincubation with piroxicam (10(-5) M), an inhibitor of arachidonic acid cyclooxygenase, suggesting prostaglandin formation as an intermediate step for cytokine stimulation of short-circuit current. In intact mucosal strips, interleukin 1 and 3 stimulated prostaglandin E2 release and elevated tissue 3',5'-cyclic adenosine monophosphate concentration. When prostaglandin E2 release from epithelial and subepithelial fractions of the mucosa by interleukin 1 was determined, increases were found only from the subepithelium. The secretory actions of cytokines appear to be mediated by arachidonic acid metabolites most likely produced by cells of the lamina propria and submucosa and may play a role in inflammatory processes in which intestinal secretion is enhanced.

Action Potentials↗
PubMed1990

Calcium mediated neurohumoral inhibition of chicken enterocyte Na influx: role of phosphatidylinositol metabolites.

Carbachol (CCH), serotonin (5HT), divalent ionophore A23187, cAMP, and certain neuropeptides, i.e. substance P (SP), inhibit the initial rate of uptake (influx) of 22Na into isolated chicken villus enterocytes. All these agents also increase cytosolic Ca. However, the increases stimulated by CCH, 5HT, and cAMP are not blocked by chelation of extracellular Ca, whereas those of A23187 and SP are. Only CCH and 5HT stimulate hydrolysis of membrane phosphoinositides to form inositol phosphates. CCH and 5HT also stimulate incorporation of [32P]-PO4 into membrane polyphosphoinositides. These studies suggest that at least three mechanisms exist to increase cytosolic Ca in chicken enterocytes and thereby inhibit Na influx. Certain neurohumoral agents such as SP open a plasma membrane permeability for Ca, permitting extracellular Ca to enter the cell down its electrochemical gradient. These agents do not stimulate phosphatidylinositol breakdown. CCH and 5HT stimulate phosphatidylinositol breakdown and via the formation of inositol trisphosphate release Ca from intracellular stores. A third mechanism exists for cAMP which mobilizes Ca from intracellular stores, but does not involve the metabolism of membrane phosphatidylinositols.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1990

Chemotactic peptide effects on intestinal electrolyte transport.

The bacterial-derived chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (fMLP) increases short-circuit current (Isc) and arachidonic acid metabolism (AAM) in rabbit ileum and distal colon. Serosal (s) or mucosal (m) addition of fMLP transiently increases Isc. Half-maximally effective dose and maximal increases in Isc were 32 nM and 84 microA/cm2 in ileum and 234 nM and 80 microA/cm2 in colon, respectively. Piroxicam, a cyclooxygenase inhibitor, diminished the Isc response by 97% in colon and 69% in ileum. Changes in Isc were dependent on Cl- and HCO3- in the bathing media. In ileum, fMLP inhibited m-to-s 36Cl- fluxes and stimulated s-to-m 36Cl- fluxes. These changes in Cl- flux were also inhibited by piroxicam. fMLP stimulated prostaglandin E2 (PGE2) release in intact tissue and in isolated subepithelial components. Increased tissue adenosine 3',5'-cyclic monophosphate levels were detected in intact tissue but not in isolated components. Previous desensitization of ileum to PGE1 inhibited fMLP stimulation of Isc in ileum by 88%. Desensitization to fMLP in ileum failed to alter the effect of PGE1 (10 microM) on Isc. In isolated microsomal membranes of ileal enterocytes, fMLP binding sites could not be demonstrated, suggesting that fMLPs action was initially mediated via stimulation of nonepithelial cell cyclooxygenase activity. The above results indicate that fMLP stimulates net secretion in both ileum and colon probably by the activation of AAM.

1-Methyl-3-isobutylxanthine↗
PubMed1990

Inhibition of Na/H exchange in avian intestine by atrial natriuretic factor.

Effects of 8-bromo-cGMP (8-Br-cGMP) and synthetic rat atriopeptin III (APIII) on sodium absorption by isolated chicken villus enterocytes and intact chicken ileal mucosa were determined. In isolated cells, both agents significantly decreased initial rates of influx of 22Na and caused a persistent decrease in intracellular pH (pHi); effects that are not additive to those caused by amiloride (10(-3) M). The ED50 for APIII was 0.3 nM. In intact mucosa, both 8-Br-cGMP (10(-4) M) and 5-(N-methyl-N-isobutyl)amiloride (MIBA) (10(-5) M) reduced JNams and JNa.net, their effects were not additive. APIII (10(-7) M) significantly increased cellular cGMP but not cAMP. Both 8-Br-cGMP (10(-4) M) and APIII (10(-7) M) stimulated a persistent increase in cytosolic calcium (Cai), which could be prevented by pretreating the cells with the cytosolic calcium buffering agent MAPTAM or with H-8, an inhibitor of cyclic nucleotide-dependent protein kinases. Furthermore, pretreatment of cells with H-8 or the calmodulin inhibitor, calmidazolium (CM), prevented the effects of 8-Br-cGMP and APIII on pHi. However, the pHi response to subsequent addition of the calcium-ionophore ionomycin was blocked only by CM and not by H-8. These data suggest that APIII and 8-Br-cGMP inhibit amiloride-sensitive Na/H exchange by increasing Cai, an event requiring activation of cGMP-dependent protein kinase.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Inhibition of cholera-toxin-stimulated intestinal secretion by CGS 9343B in rats: a specific calmodulin inhibitor.

In this study, the effect of CGS 9343B on cholera-toxin-stimulated intestinal secretion in rats was determined using in vivo isolated loops. This recently developed compound is a potent and specific inhibitor of calmodulin, but not of protein kinase C. At a luminal dose of 15 mg/kg, CGS 9343B has little effect on basal intestinal absorption but completely inhibited the secretory effects of cholera toxin. The less specific calmodulin inhibitor, trifluoperazine, has a similar antisecretory effect, but unlike with CGS 9343B, severe toxicity was noted at luminal doses of 7.5 mg/kg. In animals where two intestinal loops were created, one with cholera toxin alone and the other with CGS 9343B and cholera toxin, significant inhibition of secretion was observed in both loops, consistent with a systemic effect of this compound. Finally, both CGS 9343B and trifluoperazine inhibited choleratoxin stimulated increases in mucosal cyclic AMP content, whereas basal levels were unaffected. We conclude that CGS 9343B significantly inhibits choleratoxin-stimulated intestinal secretions, possibly by inhibition of calmodulin-dependent adenylate cyclase activity. Its lack of major toxicity at therapeutic doses makes this compound potentially useful for the treatment of enterotoxigenic diarrheal diseases.

Animals↗
PubMed1988

Effect of disodium azodisalicylate on electrolyte transport in rabbit ileum and colon in vitro. Comparison with sulfasalazine and 5-aminosalicylic acid.

Azodisalicylate, used to treat ulcerative colitis, causes diarrhea in up to 12.5% of patients. We compared the in vitro effects of azodisalicylate, sulfasalazine, and 5-aminosalicylic acid on rabbit intestinal electrolyte transport. Distal ileal mucosae mounted in Ussing chambers were exposed to varying concentrations of the drugs. Mucosal addition of azodisalicylate (greater than 5 mM) caused the greatest anion-dependent increase in short-circuit current of 83 microA/cm2 (ED50 = 0.3 mM). Isotope flux measurements suggest that azodisalicylate may stimulate predominantly electrogenic HCO3 secretion and induces net NaCl secretion. In contrast, serosal addition of azodisalicylate and sulfasalazine (greater than 5 mM) decreased short-circuit current, and 5-aminosalicylic acid had no effect. Azodisalicylate had no effect on ion transport in distal colon. The effects of azodisalicylate in ileum were not inhibited with piroxicam (an inhibitor of cyclooxygenase). Mucosal cyclic adenosine monophosphate and cyclic guanosine monophosphate levels were unchanged after ileal exposure to azodisalicylate. Azodisalicylate appears to be a mechanistically unusual secretagogue, possibly explaining the increased incidence of diarrhea seen in patients taking the drug.

Aminosalicylic Acids↗
PubMed1988

In vitro effects of the long-acting somatostatin analogue SMS 201-995 on electrolyte transport by the rabbit ileum.

We have investigated the in vitro properties of SMS 201-995, a long-acting somatostatin analogue, on electrolyte transport in rabbit ileum. Similar to native somatostatin, serosal addition of this compound inhibits electrogenic anion secretion and stimulates neutral sodium and chloride absorption. Both compounds have similar maximal effects on ion transport; however, the ED50 of SMS 201-995 (2.4 X 10(-10) M) was 60 times less than that for somatostatin. In addition, unlike somatostatin, no inherent tachyphylaxis was observed in response to SMS 201-995. The antisecretory profile of SMS 201-995 was also compared with that of epinephrine. Unlike treatment with epinephrine, pretreatment of tissues with SMS 201-995 did not directly inhibit electrogenic anion secretion stimulated by vasoactive intestinal polypeptide, calcium ionophore A23187, and bethanechol. In contrast, this agent blocked vasoactive intestinal polypeptide and bethanechol inhibition of net sodium absorption. We conclude that SMS 201-995 has several unique in vitro properties that may explain its greater biologic activity compared with that of somatostatin. Its effects on secretagogue-stimulated electrogenic anion secretion and electroneutral NaCl absorption appear to differ.

Absorption↗
PubMed1988

Differentiation of rat small intestinal epithelial cells by extracellular matrix.

The role of extracellular matrix as a determinant of intestinal cell maturation was explored by growing a normal, but immature, rat small intestinal cell line (IEC-6) on basement membrane extract from Engelbreth-Holm-Swarm (EHS) sarcoma cells (ECM). Grown on plastic or glass, these cells are relatively immature and proliferate rapidly. In contrast, cells on ECM attached more rapidly, stopped proliferating, and rapidly organized into multicellular complex structures. Ultrastructurally, cells grown on ECM displayed significantly more mitochondria, rough endoplasmic reticulum, apical microvilli, and complex golgi apparatus, consistent with greater maturity and synthetic activity. By indirect immunofluorescence, sucrase, alkaline phosphatase, and cellular apolipoprotein B were present in cells grown on ECM only. In contrast to cells grown on glass, these cells also demonstrated Na-dependent glucose absorption, a function unique to mature villus cells (7). We conclude that the basement membrane may be a key determinant of intestinal epithelial cell maturation. The development of a mature villuslike intestinal cell in vitro may have wide application for future studies of induction and regulation of intestinal maturation and function.

Animals↗