Biomedical subjects

H M Said

Publications and source records attributed to H M Said.

120 records · Page 7Linked to original sources

PubMed1984

Absorption of 5-methyltetrahydrofolate in rat jejunum with intact blood and lymphatic vessels.

UNLABELLED: Transport results from in vitro studies may not be applicable to in vivo situations. In this study, we extended our previous in vitro observations regarding the intestinal transport of 5-methyltetrahydrofolate to in vivo studies in the unanesthetized rat and examined the effect of the unstirred water layer on the absorption process. We used a well defined intestinal perfusion technique. Absorption of 0.5 and 5 microM 5-methyltetrahydrofolate proceeded in a linear manner for 40 min of perfusion at 0.31 and 1.74 nmol/100 cm per min, respectively. Absorption of 0.5 microM 5-methyltetrahydrofolate increased with increasing perfusate flow-rate from 0.5 to 2 to 4 ml/min, indicating an unstirred water layer influence on the absorption rate. Absorption of the substrate was saturable with an apparent Kt of 5.7 microM and Vmax of 3.45 nmol/100 cm per min. Absorption was pH-dependent, and was inhibited by structural analogues. In contrast to the in vitro data, addition of glucose (20 mM) to the perfusate was unnecessary for in vivo absorption to proceed. Unconjugated cholic (5 mM) and deoxycholic (1 mM) acids and the organic anion rose bengal (0.1 mM) inhibited the absorption of 0.5 microM 5-methyltetrahydrofolate when added to the perfusate. CONCLUSIONS: the results of previous in vitro studies of 5-methyltetrahydrofolate intestinal transport are applicable to in vivo situations, except that luminal glucose was found to be unnecessary in the latter. The unstirred water layer modulated the absorption of 5-methyltetrahydrofolate, while unconjugated bile acids and rose bengal inhibited it.

Animals↗

PubMed1984

Inhibitory effect of unconjugated bile acids on the intestinal transport of 5-methyltetrahydrofolate in rat jejunum in vitro.

The effect of the unconjugated bile acids, cholic, deoxycholic, chenodeoxycholic, and ursodeoxycholic acids, and of the conjugated bile acid taurocholic acid on the mucosal-to-serosal transport and tissue uptake of the naturally occurring folate derivative, 5-methyltetrahydrofolate (5-CH3H4PteGlu) was examined in everted sacs of rat jejunum. Each of the unconjugated bile acids examined inhibited the transport and tissue uptake of 5-CH3H4PteGlu in a concentration dependent manner. At low concentrations (0.01-0.1 mM) of cholic and deoxycholic acids, no structural or functional damage to the intestinal mucosa occurred and the transport of 5-CH3H4PteGlu was inhibited competitively with Ki values of 0.114 mM and 0.055 mM for cholic and deoxycholic acids, respectively. The greater inhibition of 5-CH3H4PteGlu transport by unconjugated bile acids at 1 mM can be attributed to observed structural and functional damage to the intestinal mucosa. The addition of 2 mM lecithin to the mucosal medium failed to prevent the inhibitory effect of 0.1 mM deoxycholic acid on the transport of 0.5 microM 5-CH3H4PteGlu. Compared with the effect of unconjugated bile acids, the conjugated bile acid taurocholic acid (0.01-5 mM) showed no effect on the transport and tissue uptake of 5-CH3H4PteGlu. The results of this study show that intestinal transport and tissue uptake of 5-CH3H4PteGlu are inhibited by unconjugated bile acids in a dose-dependent fashion. The clinical and physiological implications of these observations are discussed.

Animals↗

PubMed1984

Does aging affect the intestinal transport of 5-methyltetrahydrofolate?

Aging is associated with increased incidence of folate deficiency. The mechanism responsible for the deficiency is not known but could include a decrease in the intestinal absorptive capacity for folates. Therefore, we examined the intestinal absorptive capacity for folate in young and old rats. We assessed the absorption of the natural folate derivative, 5-methyltetrahydrofolate in 3- and 25-month-old rats, using jejunal everted sacs. The kinetic parameters of 5-methyltetrahydrofolate absorption disclosed apparent Kt of 7.7 and 6.54 microM and Vmax of 0.96 and 1.14 nmol/g initial tissue wet weight/30 min in young and old rats, respectively. These data do not demonstrate an aging-associated decrease in the intestinal transport capacity for 5-methyltetrahydrofolate. Therefore the deficiency of folate found in the elderly is unlikely to be due to intestinal malabsorption of folate monoglutamates. Other mechanisms must play a role in the deficiency of folate seen in the elderly.

Aging↗

PubMed1984

Inhibitory effect of unconjugated bile acids on the enterohepatic circulation of methotrexate.

The effect of unconjugated cholic and deoxycholic acids on intestinal and hepatic transport and bile secretion of methotrexate was studied using everted sacs of rat proximal jejunum and isolated perfused rat liver. Cholic and deoxycholic acids competitively inhibit the mucosal-to-serosal transport of methotrexate (Ki, 0.08 and 0.06 mM, respectively). Cholic and deoxycholic acids also decrease intestinal tissue content of methotrexate in a concentration-dependent manner. Structural and functional damage to the intestinal mucosa does not occur in tissue treated with 0.1 mM and lower concentration of deoxycholic acid as assessed by histological studies, transmural potential difference measurements and the release of the cytoplasmic marker enzyme, lactate dehydrogenase. In the isolated liver, cholic and deoxycholic acids inhibit the uptake, retention and biliary secretion of methotrexate. At 1 mM cholic and deoxycholic acids, 72 and 80% inhibition in liver uptake and 93 and 99% inhibition in bile secretion of 1 microM methotrexate are observed, respectively. These studies demonstrate that unconjugated bile acids inhibit the enterohepatic circulation of methotrexate by impairing its intestinal transport and hepatic uptake and retention and biliary secretion.

Animals↗

PubMed1984

Intestinal absorption of 5-methyltetrahydrofolate in experimental uremia.

Folate deficiency and megaloblastic anemia occur in chronic renal failure. However, the possible role of intestinal malabsorption as a cause of the reported deficiency has not been investigated. Therefore, we examined the intestinal absorption of 5-methyltetrahydrofolate in rats made uremic by subtotal nephrectomy using in vivo perfusion technique and in vitro everted sac technique. The results were compared with those obtained in a group of sham-operated rats with normal renal function. The amount of 5-methyltetrahydrofolate absorbed in vivo was significantly lower in the uremic animals as compared to the control group. In contrast, no significant difference was found in the absorption of 5-methyltetrahydrofolate in vitro in the two groups. To mimic the uremic environment, the in vitro studies were repeated using jejunal sacs from normal animals filled with either buffer solution, or sera from uremic patients before and after dialysis. Their results showed a marked suppression of 5-methyltetrahydrofolate absorption with predialysis sera and a significant improvement with post dialysis sera. We conclude that intestinal absorption of 5-methyltetrahydrofolate is impaired in uremia. The results of the in vitro experiments suggest that the observed transport defect is due to some influence of uremic environment rather than to an acquired intrinsic defect of enterocytes in uremia.

Anemia, Megaloblastic↗

PubMed1983

Cyclic adenosine-3',5'-monophosphate and folate transport in rat jejunum.

The intestinal transport of 5-methyltetrahydrofolate and pteroylmonoglutamate was examined in everted sacs of rat jejunum exposed to compounds which increase intracellular cyclic adenosine-3', 5'-monophosphate. Adenyl cyclase stimulators (hydrocortisone and prostaglandin), phosphodiesterase inhibitors (3-isobutyl-l-methylxanthine, aminophylline and papaverine), and dibutyryl adenosine-3',5'-cyclicmonophosphate added to the mucosal medium inhibit the mucosal-to-serosal transport of physiological concentrations of 5-methyltetrahydrofalate and pteroylmonoglutamate. Transport inhibition is correlated with the ability of these agents to increase cellular cyclic adenosine-3', 5'-monophosphate. The active, carrier-mediated transport system of folate compounds is highly sensitive to the increase in cyclic adenosine-3', 5'-monophosphate level, while the diffusion system is insensitive. These data indicate that the active transport system of folates is modulated by cellular cyclic adenosine-3', 5'-monophosphate.

1-Methyl-3-isobutylxanthine↗

PubMed1983

A pH-dependent, carrier-mediated system for transport of 5-methyltetrahydrofolate in rat jejunum.

The intestinal transport of the naturally occurring folate coenzyme, 5-methyltetrahydrofolate, was studied using everted sacs of rat jejunum. The study provides evidence that intestinal transport of 5-methyltetrahydrofolate is composed of two systems: 1) an active, carrier-mediated system which is demonstrable at low concentrations; and 2) a diffusion system which is demonstrable at high concentrations. The active system is characterized by: 1) saturation kinetics with Km congruent to 0.3 microM; 2) accumulation against a concentration gradient with a serosal-to-mucosal ratio of 1.8; 3) inhibition by metabolic poisons; 4) inhibition by oxidized and reduced folate analogs; 5) temperature dependence; 6) sodium dependence; 7) glucose dependence; and 8) specificity for the jejunum. These features are strongly pH-dependent, and demonstration of active transport of 5-methyltetrahydrofolate requires a buffer pH of 6, glucose in the incubation medium and a substrate concentration of less than 10(-6) M. The diffusion process is characterized by: 1) linear increase in the mucosal-to-serosal transport of 5-methyltetrahydrofolate with increasing mucosal concentration to 10(-6) M and above; 2) energy independence; 3) pH independence; and 4) temperature independence. These studies clarify the mechanism of intestinal transport of 5-methyltetrahydrofolate, show the similarities to transport of other folate compounds and provide a unified concept of intestinal folate transport.

Animals↗

PubMed1981