Pepsinogens: physiology, pharmacology pathophysiology and exercise.

Human gastric mucosa contains aspartic proteinases that can be separated electrophoretically on the basis of their physical properties into two major groups: Pepsinogen I (PGA, PGI); and Pepsinogen II (PGC, PGII). Pepsinogens consist of a single polypeptide chain with molecular weight of approximately 42,000 Da. Pepsinogens are mainly synthesized and secreted by the gastric chief cells of the human stomach before being converted into the proteolytic enzyme pepsin, which is crucial for the digestive processes in the stomach. Pepsinogen synthesis and secretion are regulated by positive and negative feed-back mechanisms. In the resting state pepsinogens are stored in granules, which inhibit further synthesis. After appropriate physiological or external chemical stimuli, pepsinogens are secreted in the stomach lumen where hydrochloric acid, secreted by the parietal cells, converts them into the corresponding active enzyme pepsins. The stimulus-secreting coupling mechanisms of pepsinogens appear to include at least two major pathways: one involving cAMP as a mediator, the other involving modification of intracellular Ca(2+)concentration. Physiological or external chemical stimuli acting through the intracellular metabolic adenyl cyclase are more effective in inducing ' de novo ' pepsinogen synthesis than those acting through intracellular Ca(2+). The activation of protein kinase C (PK-C) would appear to be involved in regulatory processes. The measurement of pepsinogens A and C in the serum is considered to be one of the non-invasive biochemical markers for monitoring peptic secretion and obtaining information on the gastric mucosa status of healthy subjects. Recently, pepsinogen measurements have been used as an effective biochemical method for evaluating and monitoring patients with gastrointestinal diseases and for checking the effects of drug treatment. The level of PGA in the serum is always high in normal gastritis, while in atrophic gastritis it is always low. In both cases the PGC level in the serum is high. In most gastrointestinal pathologies the ratio between the PGA/PGC decreases. Various reports concerning hormone and/or enzyme modification as well as gastrointestinal distress in the case of long distance exercise have been reported. It has been suggested that the origin of the gastrointestinal distress experienced by long distance runners is a transient ischaemia of the gastric mucosa; it is also suggested that a hypobaric-hypoxic environment could contribute to induce gastric mucosa necrosis. Interrelation between gastrointestinal distress, hypobaric-hypoxic environment and modifications of PGA and PGC, gastrin and cortisol was evaluated in 13 athletes after a marathon performed at 4300 m. Gastrointestinal symptoms occurred in approximately 40% of the athletes. After the race the athletes showed a significant increase of gastrin and cortisol, while the ratio between PGA/PGC decreased. No relationship was observed between gastrointestinal symptoms and hormonal changes after the race. A control group of five subjects, who had been exposed to the same environmental conditions, showed no gastrointestinal or hormonal alteration. Conversely, control subjects presented a significant decrease of cortisol related to the circadian rhythm. The same incidence of gastrointestinal symptoms at high altitude and at sea level and the absence of pathological alteration of PGA and PGC in the serum of the athletes indicates that running a marathon and living for 6 days at 4300 m does not induce gastric mucosa necrosis. Cortisol and gastrin alteration observed in the athletes at this altitude would seem to be related to an activation of the mesopontine and forebrain structures involved in the behavioural and metabolic integration of the autonomic control and arousal and psychophysical-exercise stress. 2000 Academic Press@p$hr

Antimicrobial peptides derived from pepsinogens in the stomach of the bullfrog, Rana catesbeiana.

Three antimicrobial peptides, which had strong antimicrobial activity against a broad spectrum of microorganisms, were isolated from the stomach of the bullfrog, Rana catesbeiana. Two of the antimicrobial peptides were found to be derived from the N-terminal sequences of pepsinogen A and C prosequences. The amino acid sequences of the new antimicrobial peptides, named bullfrog pepsinogen A-derived antimicrobial peptide (bPaAP) and bullfrog pepsinogen C-derived antimicrobial peptide (bPcAP), were Gly-Val-Val-Lys-Val-Ser-Arg-Leu-Lys-Gly-Glu-Ser-Leu-Arg-Ala-Arg-Leu (MW 1865.5) and Ile-Ile-Lys-Val-Pro-Leu-Lys-Lys-Phe-Lys-Ser-Met-Arg-Glu-Val-Met-Arg-A sp-His-Gly-Ile-Lys-Ala-Pro-Val-Val-Asp-Pro-Ala-Thr-Lys-Tyr (MW 3691.6), respectively. The bPaAP and bPcAP adopted 35% and 42% amphipathic alpha-helical structure in 50% trifluoroethanol, respectively, and were non-hemolytic up to a concentration of 200 microg/ml. Synthesized pepsinogen C prosequences of monkey and human, which had similar structural characteristics as bPaAP and bPcAP, also showed antimicrobial activity at concentrations of 10-200 microg/ml. The third peptide was buforin I, previously found in the stomach of the Asian toad, Bufo bufo gargarizans. These findings strongly suggest that peptides derived from the prosequences of pepsinogens, along with buforin I, may contribute to the antimicrobial function of the gastrointestinal mucosa of vertebrates, including human.

[The importance of the plasma proteins of the blood in maintaining the relative constancy of its hydrolytic properties]. / Znachenie belkov plazmy krovi v obespechenii otnositel'nogo postoianstva ee gidroliticheskikh svoistv.

Effects of hypervolemia, dehydration, activation and inhibition of urine-formation, i. v. administration of amylase and pepsinogen, experimental acute pancreatitis upon amylolytic activity of blood plasma, contents of pepsinogen in it, amylase and pepsinogen within the blood plasma protein factions, and excretion of enzymes with the urine, were studied in dogs. The data obtained suggest an important role of interconnection between amylase and pepsinogen with the plasma proteins in their renal and extrarenal excretion from the organism and in maintenance of a relative constancy of the contents and activity of enzymes in the blood. The affinity of the plasma proteins to their stains can indirectly characterise the transport capacity of the proteins in respect to amylase and pepsinogen.

[The role of the proteins of the blood plasma in the homeostasis of its amylase and pepsinogen]. / Rol' belkov plazmy krovi v gomeostaze ee amilazy i pepsinogena.

Hypervolemia and stimulation of diuresis in dogs was found to decrease the amylolytic activity of the blood plasma and pepsinogen concentration. The number of enzymes bound with blood plasma albumins decreases in hyper-fermentation. The changing connexion of the plasma albumins with the enzymes entails a renal (as well as extrarenal) extraction of the enzymes from the organism and maintenance of their relatively constant concentration and activity in the blood.

Effects of hickory-smoke condensate on development of pepsinogen 1-altered pyloric glands in rats.

Hickory-smoke condensate (HSC) is a popular food flavouring in the USA. Available data have suggested that this food additive has tumour-initiating/promoting potential. Accordingly, a commercial HSC has been investigated for its capacity to promote tumours in the rat glandular stomach using pepsinogen 1 (Pg 1)-altered pyloric glands (PAPG) as the marker of preneoplastic lesions. The development of PAPG initiated by a single intragastric administration of N-methyl-N'-nitroso-N-nitrosoguanidine was significantly increased by feeding rats a diet containing 5% HSC; no effect was observed with lower doses (1.25 or 2.5%) of HSC. The results suggest that HSC has weak tumour-promoting potential in the rat glandular stomach.

Inhibition of aspartic proteinases by propart peptides of human procathepsin D and chicken pepsinogen.

Two propart peptides of aspartic proteinases, the propart peptide of chicken pepsin and human cathepsin D, respectively, were investigated from the point of view of their inhibitory activity for a set of aspartic proteinases. These peptides display a very broad inhibitory spectrum. The strongest inhibition was observed for pepsin A-like proteinases where propart peptides can be used as titrants of active enzymes.

Organ-specific modification of tumor development by low-dose combinations of agents in a rat wide-spectrum carcinogenesis model.

The combined effects of low doses of various carcinogens and carcinogenesis modifiers on tumor development were investigated by using a wide-spectrum organ carcinogenesis model in F344 rats. These agents were administered as three groups: (1) a group of known hepatocarcinogens; (2) a group of nitroso compounds having various target organ specificities; and (3) a group of antioxidants having various inhibiting or enhancing activities depending on the target organ. Doses were used which were generally below the known effective level for the individual chemical. These groups of chemicals were administered with or without prior administration of N-diethylnitrosamine (100 mg/kg body wt., i.p.), N-methylnitrosourea (4 x 20 mg/kg body wt., i.p.) and dihydroxy-di-N-propylnitrosamine (0.1% in drinking water for 2 weeks). The hepatocarcinogen group in combination with various nitroso compounds increased the incidences of liver hyperplastic nodules and hepatocellular carcinomas. In contrast, incidences were clearly reduced when the hepatocarcinogens and/or the nitroso compounds were administered in combination with the antioxidants. For the urinary bladder, the combination with nitroso compounds and antioxidants enhanced cancer development, and the addition of hepatocarcinogens further increased tumorigenesis. For the glandular stomach, additive effects on the numbers of pepsinogen isozyme 1-altered pyloric glands, a putative preneoplastic lesion, were produced by the combination treatment of antioxidants and the nitroso compounds. No synergistic effects on tumor development were seen in other organs. The results of the present study demonstrated that combinations of various compounds at low doses can additively or synergistically exert either enhancing or inhibitory effects on the development of preneoplastic and neoplastic lesions in different organs in a single model having a wide spectrum of organ effects.

Effects of bile acids on development of pepsinogen-altered pyloric glands in rats.

The effects of dietary bile acids on the development of pepsinogenaltered pyloric glands (PAPG) were examined. Male WKY/NCrj rats were given a single dose of 160 mg/kg body wt. of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) by gastric intubation and fed basal diet containing 0.3% sodium taurocholate (Na-TC), 0.3% sodium cholate (Na-C), 0.3% sodium glycocholate (Na-GC), 0.3% sodium tauroglycocholate, 0.3% sodium deoxycholate, 0.1% chenodeoxycholic acid or 0.5% lithocholic acid for 14 weeks. All rats also received 1 ml of saturated NaCl solution 4 times by i.g. intubation. At the end of week 16, the animals were killed, and the number of PAPG per cm of mucosal length was determined immunohistochemically. Na-TC, Na-C and Na-GC significantly increased the number of PAPG over the control value, suggesting that they may have activity to promote gastric carcinogenesis.

A comparative study of gastric secretory stimulants in conscious dogs.

1. In conscious dogs with gastric fistulae, Heidenhain pouches and Thomas duodenal fistulae, pentagastrin was found to be a more potent peak acid and pepsin stimulant in both innervated stomach and vagally denervated pouch than methacholine chloride. 2. Slopes of curves relating response to the logs of molar doses of pentagastrin and methacholine, at peak secretion, did not differ significantly. The maximal pentagastrin stimulated secretion from the pouch was smaller than that for methacholine; from the fistula they did not differ. 3. Ganglionic blockade depressed methacholine stimulated peak acid secretion, but did not affect pentagastrin stimulated acid secretion. Dose-response curves for methacholine-induced acid and pepsin secretion at the perigee did not differ from those obtained with ganglionic blockade. 4. Ganglionic blockade depressed pepsin secretion from the fistula whether stimulated with pentagastrin or methacholine. Pouch pepsin secretion was small and no difference between curves was seen.

Regulation of interleukin-1 production in murine macrophages and human monocytes by a normal physiological constituent.

The in vitro production of large quantities of interleukin-1 (IL-1) in mouse peritoneal exudate macrophages and human peripheral blood monocytes is possible through the use of the proteolytic enzyme pepsin and its zymogen pepsinogen. Equal amounts of IL-1 are generated by pepsin in the absence or presence of polymixin B. The addition of pepsin or pepsinogen had no effect on the proliferation of C3H/HeJ thymocytes to the plant mitogen phytohemagglutinin. Pepsin and pepsinogen are present in significant quantities in immune cells and the plasma. Although little is known concerning the physiological role of pepsin and pepsinogen outside of the gastrointestinal system, it may be proposed that the in vivo production of IL-1 may in part be regulated by the cellular and plasma concentrations of pepsin and pepsinogen.

[Effect of aminazin on hydrolase activity of the blood and their excretion by the kidneys]. / Vliianie aminazina na aktivnost' gidrolaz v krovi i ikh vydelenie pochkami.

Chronic experiments in dogs revealed that 0.5 mg/kg of aminasin did not affect the content of pepsinogen, amylase and lipase in the peripheral blood plasma but somewhat increased the variability of the blood pepsinogen and lipase. 2 mg/kg of aminasin decreased the pepsinogen content in the blood; 5 mg/kg decreased the amylase level as well. Aminasin decreased the renal excretion of pepsinogen in direct proportion to its dosage. 5 mg/kg of aminasin decreased the renal excretion of amylase as well, with no considerable effect on the renal excretion of lipase. Aminasin (5 mg/kg) delayed the period of restoration of the blood level of amylase and pepsinogen administered parenterally, and intensified the disenzymemia caused by pepsinogen administration; the renal excretion of enzymes was decreased. The central nervous mechanisms responding to the above doses of aminasin are concluded to play a major role in maintaining the homeostasis of hydrolytic enzymes in the blood.

Inhibition of pepsin by analogues of pepsinogen-(1-12)-peptide with substitutions in the 4-7 sequence region.

Derivatives of the 1-12 sequence of pig pepsinogen were prepared by solid-phase peptide synthesis. The three derivatives contain substitutions in the 4-7 region of the 1-12 sequence. Glycine was used to replace the hydrophobic residues -Val-Pro-Leu-Val- in pairs. After cleavage and purification, the synthetic peptides were compared with a synthetic peptide of the native sequence, prepared at the same time, with respect to their ability to inhibit the pepsin-catalysed clotting of milk. Inhibitory potency, determined from plots of percentage inhibition versus concentration of synthetic peptide, is inversely correlated with the substitution of glycine residues for the hydrophobic residues. Therefore the equilibrium inhibition of pepsin by these peptides is dominated by the hydrophobic nature of the 4-7 sequence region.

[Anabolic effects of parenterally administered gastric gland hydrolases]. / Anabolicheskie vliianiia parenteral'no vvodimykh gidrolaz pishchevaritel'nykh zhelez.

In rats, i.v. administration of amylase, pepsinogen, and tripsinogen in microdoses increased the incretion of radiomethyonin from the blood into the tissue proteins of some organs. The incretion of pepsinogen stimulated the protein-production function of pancreas, tripsinogen stimulated the function of stomach glands, amylase--the function of the liver and the small intestine mucosa. The labeled pepsinogen 125I concentrated mainly in secretory organs.

Improved pepsin inhibitor derived from activation peptide 1-16 of porcine pepsinogen.

The peptide Leu-Val-Lys-Val-Pro-Leu-Val-Arg-Lys-Lys-Ser-Leu-Arg-Gln-Asn-Leu, a known pepsin inhibitor, is derived from the first 16 amino acids of porcine pepsinogen. It was prepared from the activation mixture and was modified by guanidination of its three lysine residues to form homoarginine residues. The modified peptide is a better pepsin inhibitor than the native peptide; for 50% inhibition of the milk clotting action of pepsin at pH 5.3, the molar ratio of peptide to pepsin required is 9 for the native inhibitor and only 2 for the guanidinated inhibitor. The dissociation constants (k1) of the inhibitor-pepsin complexes are 7 X 10(-8) and 1.4 X 10(-8) M for the native and guanidinated peptides, respectively. The guanidinated peptide is more resistant to digestion by pepsin at pH 3.5. The native and modified peptides partially protect pepsin from inactivation at pH 7. Stepwise removal of the amino-terminal Leu-Val-Har residues from the guanidinated inhibitor by Edman degradation decreases the pepsin-inhibiting activity only slightly at the first step, but markedly at the second and third steps. Thus, all of the amino-terminal sequence except the leucine residue is necessary for full activity.

Chemical modification of a pepsin inhibitor from the activation peptides of pepsinogen.

The peptide comprising the first 16 amino acids of porcine pepsinogen, prepared from the activation mixture, has been modified by guanidination of its three lysine residues to form homoarginine residues. The modified peptide is a better pepsin inhibitor than is the native peptide; for 50% inhibition of the milk-clotting action of pepsin at pH 5.3, the molar ratio of peptide to pepsin required is 9 for the native inhibitor and only 2 for the guanidinated inhibitor. Stepwise removal by Edman degradation of the amino-terminal Leu-Fal-Homoarg residues from the guanidinated inhibitor decreased the activity slightly at the first step and markedly at the second and third steps. Thus, all of the amino-terminal sequence except the leucine residue is necessary for full activity.

A restaining method to restore fluorescence in faded preparations of tissues treated with the indirect immunofluorescence technique.

We report a restainin method for restoring fluorescence in paraffin-embedded tissue sections previously treated with the indirect immunofluorescence technique. Antisera to gastrin and group II pepsinogens were used. Fluorescence was restored in completely faded sections retrived from storage files, as well as in sections that had faded partially either with exposure to fluorescence microscope illumination or after counterstaining with hematoxylin and eosin.