Capsaicin Induces Cystatin S-like Substances in Submandibular Saliva of the Rat

Abstract

Irritating dietary substances such as tannin and papain have been reported to alter the morphology of salivary glands and their secretions. Such alterations can be one line of protection from toxic or irritating substances in food. We investigated the effects of dietary capsaicin (a pungent ingredient of hot red pepper) on the rat submandibular gland and its secretions. Several groups of animals were offered either control diets or diets containing capsaicin (from 0.0001 to 0.1%) for seven days. Higher concentrations suppressed food consumption for two days, after which only the highest concentration continued to reduce intake. The relative weight of the salivary glands in capsaicin-diet groups increased in a dose-dependent fashion, and new proteins appeared in the submandibular saliva. Chromatographic and electrophoretic properties of these proteins were identical or similar to those of isoproterenol-induced proteins. After affinity chromatography of the new protein fraction on a Cm-papain Sepharose 4B column, SDS-electrophoresis of the eluate revealed three major bands (15,500, 16,500, and 28,000 kDa). Hydrolysis of N-benzoyl-D,L-arginine-p-nitroanilide by papain (a cysteine protease) decreased in the presence of the new protein fraction, suggesting that these proteins have cystatin-like activity (inhibition of cysteine protease). Denervation of the glossopharyngeal nerve suppressed induction of these proteins. The results suggest that dietary capsaicin induces cystatin S-like substances in submandibular saliva by stimulating the reflex arc involving the glossopharyngeal nerve. These proteins likely facilitate ingestion of diets containing the irritating substance.

Keywords

capsaicin cystatin S-like substances submandibular saliva

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References

Ainsworth A. , Hall P. , Wall PD , Allt G. , Mackenzie ML , Gibson S. , Polak JM (1981). Effects of capsaicin applied locally to adult peripheral nerve. II. Anatomy and enzyme and peptide chemistry of peripheral nerve and spinal cord. Pain 11:379-388.

Anastasi A. , Brown MA , Kembhavi AA , Nicklin MJ , Sayers CA , Sunter DC , et al. (1983). Cystatin, a protein inhibitor of cysteine proteinases. Improved purification from egg white, characterization, and detection in chicken serum. Biochem j 211:129-138.

Beil W. , Staar U. , Sewing K. -F (1992). Pantoprazole: a novel H+ /K+-ATPase inhibitor with an improved pH stability. Eur J Pharmacol 218:265-271.

Bobek LA , Levine MJ (1992). Cystatins-Inhibitors of cysteine proteinases. Crit Rev Oral Biol Med 3:307-332.

Duner-Engstrom M. , Fredholm BB , Larsson O. , Lundberg JM , Saria A. (1986). Autonomic mechanisms underlying capsaicin induced oral sensations and salivation in man. J Physiol 373:87-96.

Fazekas A. , Vindisch K. , Posch E. , Gyorfi A. (1990). Experimentally-induced neurogenic inflammation in the rat oral mucosa. J Periodont Res 25:276-282.

Glendinning JI (1992). Effect of salivary proline-rich proteins on ingestive responses to tannic acid in mice. Chem Senses 17:1-12.

Green I. (1989). Capsaicin sensitization and desensitization on the tongue produced by brief exposure to a low concentration. Neurosci Lett 107:173-178.

Kawada T. , Watanabe T. , Takaishi T. , Tanaka T. , Iwai K. ( 1986). Capsaicin-induced β-adrenergic action on energy metabolism in rats: influence of capsaicin on oxygen consumption, the respiratory quotient, and substrate utilization. Proc Soc Exp Biol Med 183:250-256.

10.

Ki P. , Negulesco JA , Murnane M. (1982). Decreased total serum, myocardial and aortic cholesterol levels following capsaicin treatment. IRCS Med Sci Libr Compend 10:446-447.

11.

Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.

12.

Lowry OH , Rosebrough NJ , Farr AL , Randall RJ (1951). Protein measurement with the folin phenol reagent . J Biol Chem 193:265-275.

13.

Lundberg JM (1989). Peptidergic control of the autonomic regulation system in the orofacial region. Proc Finn Dent Soc 85:239-250.

14.

Matsuo T. , Yoshioka M. , Suzuki M. (1996). Capsaicin in diet does not affect glycogen contents in the liver and skeletal muscle of rats before and after exercise. J Nutr Sci Vitaminol 42:249-256.

15.

Mehansho H. , Carlson DM (1983). Induction of protein and glycoprotein synthesis in rat submandibular glands by isoproterenol. J Biol Chem 258:6616-6620.

16.

Mehansho H. , Hagerman A. , Clements S. , Butler L. , Rogler J. , Carlson DM (1983). Modulation of proline-rich protein biosynthesis in rat parotid glands by sorghums with high tannin levels. Proc Natl Acad Sci USA 80:3948-3952.

17.

Mehansho H. , Clements S. , Sheares BT , Smith S. , Carlson DM (1985). Induction of proline-rich glycoprotein synthesis in mouse salivary glands by isoproterenol and by tannins. J Biol Chem 260:4418-4423.

18.

Naito Y. , Suzuki I. , Hasegawa S. (1992). Induction of cystatin S in rat submandibular glands by papain. Comp Biochem Physiol 102(B):861-865.

19.

Ninomiya Y. , Kajiura H. , Naito Y. , Mochizuki K. , Katsukawa H. , Torii K. (1994). Glossopharyngeal denervation alters responses to nutrients and toxic substances. Physiol Behav 56:1179-1184.

20.

Nishimura T. , Ishibashi K. , Abe K. (1991). Isolation of three forms of cystatin from submandibular saliva of isoproterenol-treated rats, its properties and kinetic data. Biochem Biophys Acta 1077:346-354.

21.

Sambaiah K. , Satyaranayana MN (1982). Lipotrope-like activity of red pepper. J Food Sci Technol 19:30-31.

22.

Shaw PA , Cox JL , Barka T. , Naito Y. (1988). Cloning and sequencing of cDNA encoding a rat salivary cysteine protease inhibitor inducible by p-adrenergic agonist. J Biol Chem 263:18133-18137.

23.

Shaw PA , Barka T. , Woodin A. , Schacter BS , Cox JL (1990). Expression and induction by β-adrenergic agonists of the cystatin S gene in submandibular glands of developing rats. Biochem J 265:115-120.