Parameters of oxidative stress in saliva from patients with aggressive and chronic periodontitis.

OBJECTIVES: Free radicals play an important role in the onset and progression of many diseases. The aim of this study was to investigate the contribution of oxidative stress in the pathology of aggressive (AgP) and chronic (CP) periodontitis and its relation with the clinical periodontal status. METHODS: Eighty subjects were divided into two groups: 20 patients with AgP and 20 patients with CP with their 20 corresponding matched controls, based on clinical attachment loss (CAL), probing pocket depth (PPD), and bleeding on probing (BOP). Saliva reactive oxygen species (ROS), lipid peroxidation, and non-enzymatic antioxidant defences were measured by luminol-dependent chemiluminescence assay, as thiobarbituric acid-reactive substances (TBARs) and total radical-trapping antioxidant potential (TRAP), respectively. Pearson's correlation and multivariate analysis were used to determine the relationship between ROS and TBARs and the clinical parameters. RESULTS: ROS and TBARs were increased in AgP while TRAP was decreased, comparing with CP. In AgP, a strong and positive correlation was observed between ROS and TBARs and they were closely associated with CAL and PPD. DISCUSSION: In AgP, but not in CP, oxidative stress is a high contributor to periodontal pathology and it is closely associated with the clinical periodontal status.

Comparison of salivary levels of mucin and amylase and their relation with clinical parameters obtained from patients with aggressive and chronic periodontal disease.

OBJECTIVE: Salivary mucin and amylase levels are increased in patients with chronic periodontitis (CP). Due to the fact that aggressive periodontitis (AgP) not only differs from chronic periodontitis in terms of its clinical manifestation, the aim of this study was to compare salivary mucin and amylase levels and their relation to the clinical parameters of patients with aggressive periodontitis with that of patients with chronic periodontitis. MATERIAL AND METHODS: Eighty subjects were divided into two groups: 20 patients with AgP and their 20 matched controls and 20 patients with CP and their 20 matched controls, based on clinical attachment loss (CAL), probing pocket depth (PPD) and bleeding on probing (BOP). Whole unstimulated saliva was obtained and mucin, amylase and protein were determined by colorimetric methods. Pearson's correlation analysis was used to determine the relationship between salivary mucin, amylase and protein levels and the clinical parameters. RESULTS: Salivary mucin, amylase and protein levels were increased in patients with AgP and CP but there were no differences between them or between control groups. Pearson's correlation analysis, determined in the entire subjects studied, showed a positive and significant correlation of mucin, amylase and proteins with CAL and PPD and a negative correlation with the flow rate. When Pearson's correlation analysis was carried out in each group separately, Fisher's z transformation showed no significant difference between both groups. CONCLUSION: Comparison of the salivary levels of mucin, amylase and protein and their relationship with clinical parameters of AgP patients with that of CP patients revealed no differences between both groups.

Comparison of salivary levels of mucin and amylase and their relation with clinical parameters obtained from patients with aggressive and chronic periodontal disease

Objective Salivary mucin and amylase levels are increased in patients with chronic periodontitis (CP). Due to the fact that aggressive periodontitis (AgP) not only differs from chronic periodontitis in terms of its clinical manifestation, the aim of this study was to compare salivary mucin and amylase levels and their relation to the clinical parameters of patients with aggressive periodontitis with that of patients with chronic periodontitis. Material and Methods Eighty subjects were divided into two groups: 20 patients with AgP and their 20 matched controls and 20 patients with CP and their 20 matched controls, based on clinical attachment loss (CAL), probing pocket depth (PPD) and bleeding on probing (BOP). Whole unstimulated saliva was obtained and mucin, amylase and protein were determined by colorimetric methods. Pearson's correlation analysis was used to determine the relationship between salivary mucin, amylase and protein levels and the clinical parameters. Results Salivary mucin, amylase and protein levels were increased in patients with AgP and CP but there were no differences between them or between control groups. Pearson's correlation analysis, determined in the entire subjects studied, showed a positive and significant correlation of mucin, amylase and proteins with CAL and PPD and a negative correlation with the flow rate. When Pearson's correlation analysis was carried out in each group separately, Fisher's z transformation showed no significant difference between both groups. Conclusion Comparison of the salivary levels of mucin, amylase and protein and their relationship with clinical parameters of AgP patients with that of CP patients revealed no differences between both groups. .

Association between Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis in subgingival plaque and clinical parameters, in Argentine patients with aggressive periodontitis.

BACKGROUND: Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) have been associated with aggressive (AgP) and chronic periodontitis. OBJECTIVE: The aim of this study was to evaluate the levels of Aa and Pg in gingival crevicular fluid (GCF) of patients with AgP and its relation with clinical parameters. DESIGN: Sixteen females and fourteen males with clinical diagnosis of AgP aged 17-23 years and their match's controls, were included in this study. Clinical recording concerning probing pocket depth, clinical attachment level, plaque index and gingival bleeding index were performed at baseline, 30 and 60 days after baseline. After clinical examination GCF samples were analyzed for Aa and Pg with a real-time polymerase chain reaction technique. Patients group was treated with a combined of mechanical and oral antibiotic therapy (doxycycline 100 mg/day, during 21 days). A multivariate analysis was used to determine the relationship between Aa and Pg counts with clinical parameters. RESULTS: GCF from all subjects was positive for Aa and PG. In controls Pg concentration was higher than Aa (Pg: 42,420 ± 3,034 copies/ml; Aa: 66.6 ± 5.4 copies/ml p < 0.001) while in patients both microbes showed the same concentration (Aa: 559,878 ± 39,698 Pg: 572,321 ± 58,752). A significant and positive correlation was observed between counts of Aa and Pg (R square: 0.7965, p < 0.0001). Female showed more counts/ml. Aa might be closely associated with clinical parameters while Pg did not. At 30 and 60 days Aa counts in patients were similar to controls while Pg counts were equal to baseline. However, in spite of Pg presence a clinical improvement was observed in all patients. CONCLUSIONS: In our population the presence of Aa may be associated with AgP while Pg may be in GCF as an opportunistic pathogen which might caused disease when the ecological balance was favorable.

Total salivary nitrates and nitrites in oral health and periodontal disease.

It is well known that nitrites are increased in saliva from patients with periodontal disease. In the oral cavity, nitrites may derive partly from the reduction of nitrates by oral bacteria. Nitrates have been reported as a defence-related mechanism. Thus, the aim of the present study was to determine the salivary levels of total nitrate and nitrite and their relationship, in unstimulated and stimulated saliva from periodontal healthy subjects, and from patients with chronic periodontal disease. Nitrates and nitrites were determined in saliva from thirty healthy subjects and forty-four patients with periodontal disease. A significant increase in salivary nitrates and nitrites was observed. Nitrates and nitrites concentration was related to clinical attachment level (CAL). A positive and significant Pearson's correlation was found between salivary total nitrates and nitrites. Periodontal treatment induced clinical improvement and decreased nitrates and nitrites. It is concluded that salivary nitrates and nitrites increase, in patients with periodontal disease, could be related to defence mechanisms. The possibility that the salivary glands respond to oral infectious diseases by increasing nitrate secretion should be explored further.

Salivary IL-1ß and PGE2 as biomarkers of periodontal status, before and after periodontal treatment.

AIM: Interleukin-1ß (IL-1ß) and prostaglandin E2 (PGE2 ) are key inflammatory mediators involved in periodontitis. The purpose was to compare their salivary concentrations in relation to periodontal status and their changes after periodontal treatment, to determine their use as non-invasive diagnostic tools. MATERIALS AND METHODS: In this study, 74 subjects grouped in periodontally healthy, mild, moderate and severe periodontitis, according to clinical attachment level (CAL) and probing pocket depth (PPD) served as participants. IL-1ß and PGE2 were determined in unstimulated whole saliva by enzyme-linked-immunosorbent assay (ELISA). RESULTS: Interleukin -1ß increased with the severity of periodontitis with a large effect size in prediction of CAL (η(2)  = 0.35, p = 0.0001). PGE2 showed an increment in mild periodontitis and another in moderate. A significant effect size was also found between PGE2 and PPD (η(2)  = 0.12, p = 0.003). Both mediators decreased after periodontal treatment. With a selected threshold of 212 pg/ml, salivary IL1-ß predicted periodontitis with 78% sensitivity and 100% specificity. With a selected threshold of 121 pg/ml, salivary PGE2 predicted periodontitis with 78% sensitivity and 91% specificity. CONCLUSION: The high sensitivity and specificity of salivary IL-1ß and PGE2 in identifying periodontitis suggest a potential use as biomarkers for diagnosis of periodontitis presence and severity.

Influence of experimental periodontitis on cholinergic stimulation of K⁺ release in rat parotid glands.

In a rat model of experimental periodontitis it was investigated whether the presence of the inflammatory disease induced changes in carbachol-induced fluid secretion in parotid glands, by monitoring potassium release. The potency of carbachol, to induce K⁺ release, was higher in parotid glands from rats with experimental periodontitis. The antagonist with higher affinity for M3 muscarinic acetyl-choline receptor subtype, 4-DAMP (selectivity: M1=M3), was more potent in inhibiting K⁺ release in periodontitis rats while the antagonist with a muscarinic M1-receptor-selective profile (selectivity: M1>M3), pirenzepine, was more potent in control rats. Competition binding assays showed that both, M1 and M3 muscarinic acetyl-choline receptor subtypes are expressed in membranes of parotid glands. The K(i) of 4-DAMP was decreased in parotid glands from rats with experimental periodontitis while the Ki of pirenzepine was increased. The effect of periodontitis was reverted by the inhibition of the cyclooxygenase activity through indomethacin treatment (100 mg/k ip, 4 days). It was concluded that periodontitis could induce changes in muscarinic acetyl-choline receptor subtypes expression with a preferential increase of M3 subtype, resulting in increased K⁺ released in response to carbachol and in a greater potency of 4-DAMP. These findings agree with the fact that a decrease of fluid secretion is not a condition of patients with periodontal disease.

Enhancement of carbachol-induced amylase secretion in parotid glands from rats with experimental periodontitis.

OBJECTIVE: In a previous study we observed that parotid glands from rats with experimental periodontitis showed an increase in basal amylase release as a result of an increase in cAMP accumulation induced by PGE(2) production. The aim of this work was to study whether this change in amylase release influences the secretory effect of carbachol. DESIGN: Experimental periodontitis was induced through placing a black thread around the cervix of the two lower first molars. Experiments were done 22 days after ligature induced periodontitis. Amylase release was evaluated in vitro and determined using a colorimetric method which uses starch as substrate. RESULTS: The effect of carbachol was increased in parotid glands from periodontitis rats. The effect of 10(-6)M carbachol was inhibited by 4-DAMP (10(-6)M), U-73122 (5 × 10(-6)M) and trifluoperazine (5 × 10(-6)M) in both groups. No changes were observed in the binding sites and affinity in parotid membranes from rats with experimental periodontitis. The inhibition of the adenylyl cyclase and the cyclooxygenase induced a right shift of the carbachol concentration-response curve in periodontitis group whilst the opposite effect was observed in control group in the presence of db-cAMP and PGE(2). CONCLUSIONS: Parotid glands from rats with experimental periodontitis release more amylase in response to carbachol suggesting an interaction between Ca(2+) and cAMP in the fusion/exocytosis step of secretory vesicles.

Experimental periodontitis induces a cAMP-dependent increase in amylase activity in parotid glands from male rats.

It is known that subjects with periodontitis show enhanced amylase concentration in saliva. Our purpose was to analyze the release of amylase in parotid glands from rats with experimental periodontitis and controls. We present evidence that periodontitis induces an increase in resting amylase activity and release without changes in isoproterenol-induced amylase secretion. Changes in amylase were reverted by the inhibition of the adenylyl cyclase by SQ 22536, the cyclooxygenase type 1 by FR 122047 and by blocking the vasoactive intestinal peptide (VIP) receptor with VIP 6-28. Parotid glands from rats with periodontitis showed an increase in cAMP levels that was also reverted in the presence of SQ 22536, FR 122047 and VIP 6-28. We concluded that both PGE(2) and VIP are produced in parotid glands from rats with periodontitis and, by activating their own receptors in acinar cells, induce cAMP accumulation leading to an increase in amylase basal secretion.

Increased leukotriene concentration in submandibular glands from rats with experimental periodontitis.

OBJECTIVE AND DESIGN: In the present study, we investigated the relation between the inflammatory mediators such as nitric oxide, prostaglandins, and cysteinyl-leukotrienes with mucin release and the sympathetic system in submandibular glands from rats with experimental periodontitis. MATERIALS OR SUBJECTS: Submandibular glands from rats with experimental periodontitis. TREATMENT: For the first experiment, rats were treated with hydrocortisone sc, 1 mg/kg for 3 days. All other experiments were carried out in isolated submandibular glands from untreated rats. Submandibular glands were treated with cysteinyl-leukotrienes, isoproterenol, NDGA, FPL 55712, L-NMMA, Nio, Nz, AMG, indomethacin, DuP 697 and atenolol. METHODS: Nitric oxide synthase activity, prostaglandin and cysteinyl-leukotriene productions and mucin secretion were determined. The Newman-Keuls statistical test was applied after analysis of variance. RESULTS: In rats with periodontitis hydrocortisone-induced a 36.6% (P < 0.05) decrease in mucin release. Only cysteinyl-leukotriene production was increased in rats with ligature (79.2%, P < 0.001). Either the inhibition of cysteinyl-leukotriene production or the block of leukotriene receptor abolished the increase in mucin secretion by 25.6% (P < 0.05) and 37% (P < 0.01), respectively, in glands from rats with ligature. On the other hand, the presence of cysteinyl-leukotrienes in the incubation medium induced mucin release from submandibular glands. Atenolol diminished by 24% (P < 0.05), the increase in cysteinyl-leukotrienes observed in rats with periodontitis. Besides, isoproterenol induced cysteinyl-leukotriene production in both groups. CONCLUSION: In submandibular glands from rats with periodontitis, the increment in mucin release and cysteinyl-leukotrienes production are related events and both are associated with the sympathetic system.

Mucinas salivales: estructura química, mecanismos de liberación y participación en la defensa no inmunológica de la cavidad bucal / Salivary mucins: chemical structure, release mechanisms and participation in the non-immunological defense of the oral cavity

En este artículo se describen: 1) las características físico-químicas de las mucinas salivales, denominadas MG1 y MG2. 2) El mecanismo de secreción por estimulación simpática y parasimpática. 3) La distinta participación de MG1 y MG2 tanto en la actividad deglutoria como en los mecanismos de defensa de la cavidad bucal, en relación con sus propiedades físico-químicas. 4) El rol de las mucinas salivales en la protección de la mucosa del tracto gastrointestinal. 5) La relación entre las mucinas saliales y las patologías de la cavidad bucal.

Mucinas salivales: estructura química, mecanismos de liberación y participación en la defensa no inmunológica de la cavidad bucal / Salivary mucins: chemical structure, release mechanisms and participation in the non-immunological defense of the oral cavity

En este artículo se describen: 1) las características físico-químicas de las mucinas salivales, denominadas MG1 y MG2. 2) El mecanismo de secreción por estimulación simpática y parasimpática. 3) La distinta participación de MG1 y MG2 tanto en la actividad deglutoria como en los mecanismos de defensa de la cavidad bucal, en relación con sus propiedades físico-químicas. 4) El rol de las mucinas salivales en la protección de la mucosa del tracto gastrointestinal. 5) La relación entre las mucinas saliales y las patologías de la cavidad bucal.(AU)

Beta-adrenoceptor alterations coupled with secretory response and experimental periodontitis in rat submandibular glands.

In this paper we have studied the influence of a well-established rat model of periodontitis on resting and adrenergic-stimulated mucin secretion from rat submandibular glands. The selective beta(1)-receptor subtype agonist, dobutamine, induced mucin secretion while the selective beta(2)-, alpha(1)- and alpha(2)-agonists, soterenol, phenylephrine and clonidine, respectively, did not. In rats subjected to ligature-induced periodontitis mucin release, under unstimulated conditions (basal values), was significantly increased. This increment was abolished in the presence of propranolol and atenolol. Isoproterenol, concentration-dependent, increased mucin release in control and in ligature-induced periodontitis rats. Maximal effect of isoproterenol was decreased in rats with ligature while EC(50) was increased. Neither, the inhibition of NOS by l-NMMA nor the inhibition of COX by indomethacin could revert the effect of ligature on mucin release under unstimulated and isoproterenol-stimulated conditions. The inhibition of adenylyl cyclase by SQ 22536 resulted in a right shift of isoproterenol concentration-response curves in both groups, control and with ligature and returned basal values of rats with ligature to control ones. beta-Receptor population was decreased in submandibular gland membranes from rats with ligature without changes in affinity. Potencies of the beta-receptor antagonists in the competition studies were similar in both groups under study, control and with ligature. We conclude that in rats subjected to ligature-induced periodontitis unstimulated mucin secretion is increased. The increment seems to be due to an activation of the sympathetic system since it is inhibited by the beta-adrenoceptors antagonists and by the inhibition of the adenylyl cyclase. We can speculate that inflammatory mediators from the experimental periodontitis could be involved in the mechanism underlying the activation of the sympathetic system.

Signaling pathways involved in pilocarpine-induced mucin secretion in rat submandibular glands.

We have studied the signaling pathways involved in pilocarpine-induced mucin release in rat submandibular slices. Pilocarpine produced a significant increment of PGE2 levels and a positive (r=0.8870) and significant (p=0.0077) correlation between PGE2 production and mucin released was determined. The participation of PGE2 was confirmed by the use of indomethacin (indo) and of acetyl salicylic acid (ASA), cyclooxygenase inhibitors, which inhibited pilocarpine-induced mucin release. The muscarinic receptors involved in the regulation of mucin release were identified as M1 and M4 by the use of the selective acetylcholine receptors (mAChR) antagonists, pirenzepine, AF-DX 116, 4-DAMP and tropicamide. The secretory process was dependent on both, intracellular and extracellular calcium pools since it was inhibited by thapsigargin and verapamil. Cyclic AMP, nitric oxide synthase and PKC also participated in pilocarpine-induced mucin release. It is concluded that pilocarpine, by activation the M1 and M4 mAChR subtypes induces an increase of intracellular Ca2+ concentration ([Ca2+]I) and elevates cAMP levels, which in turn stimulates COX, PKC and NOS and promotes mucin exocytosis. PGE2 released induces cAMP accumulation which, together with PKC are involved in the PGE2 increased Ca2+/cAMP-regulated exocytosis. Thus, cAMP accumulation induced by cholinergic stimulation is, in part, the result of PGE2 production.

An overview of autonomic regulation of parotid gland activity: influence of orchiectomy.

The parotid gland participates in the digestive process by providing fluid, electrolytes and enzymes that facilitate the onset of digestion. Neurotransmitters, hormones and biologically active peptides regulate its activity. The autonomic system is the main regulatory mechanism of the gland. Sympathetic stimulation induces amylase release through beta(1)-receptor activation and few fluid secretion by alpha(1)-receptor activation. The parasympathetic system controls basal activity of the gland acting on M(1) and M(3) muscarinic acetylcholine receptors and induces the secretion of fluid saliva rich in electrolytes through the modulation of ion channels and the Na(+)-K(+)-ATPase activity. In addition, its activation induces amylase release. The mechanisms involved in amylase secretion by isoproterenol and carbachol, as well as the mechanism of the cholinergic regulation of Na(+)-K(+)-ATPase activity and the changes observed after orchiectomy, are the scope of this review.

Effects of castration on cannabinoid cb receptor expression and on the biological actions of cannabinoid in the parotid gland.

In the present study, we examined whether cannabinoid receptor expression and the effects of receptor stimulation vary as a function of gonadal status in a peripheral tissue, namely the male rat parotid gland. Four groups of male rats were studied: gonadal intact, castrated, castrated testosterone (1 mg/100 g bodyweight) treated and gonadal intact testosterone treated. 2. The results showed that the density of CB(1) receptors decreased after castration and that receptor density was restored to control values after testosterone treatment. This decrement was associated with a decrease of anandamide (10(-10) to 10(-5) mol/L)-induced cAMP accumulation and amylase release without changes in the anandamide-induced inhibition of Na(+)/K(+)-ATPase activity. 3. Castration did not modify either the subtype of cannabinoid receptor involved in the actions of anandamide or drug affinity for the receptor. 4. The mechanism underlying anandamide-induced cAMP accumulation, amylase release and inhibition of Na(+)/K(+)-ATPase activity, namely through the activation of adenylyl cyclase, was the same in control and castrated rats. 5. Basal cAMP accumulation, amylase release and Na(+)/K(+)-ATPase activity were not altered by castration. 6. Castration had no effect on the concentration of total protein. 7. It can be concluded that CB(1) cannabinoid receptor expression is regulated by testosterone in male rat parotid gland and this has functional implications for cAMP accumulation and amylase release.

Expression and biological effects of CB1 cannabinoid receptor in rat parotid gland.

Experiments were designed to determine whether cannabinoids affect salivary gland function. For this purpose, the effect of anandamide on cAMP accumulation, amylase release and Na+-K+-ATPase activity was studied in rat parotid glands. Anandamide induced a concentration-dependent increase in cAMP and led to amylase release but inhibited Na+-K+-ATPase activity. These effects were blocked by the CB1 cannabinoid receptor antagonist, AM281. The inhibition of adenylyl cyclase activity by SQ 22536 impaired amylase release and Na+-K+-ATPase inhibition. The effect of anandamide on cAMP accumulation significantly correlated with its action either on amylase release or on Na+-K+-ATPase activity. Such correlation strongly supports the view that the effect of anandamide on amylase release and Na+-K+-ATPase activity is the result of cAMP accumulation. The relative potencies of the CB1 cannabinoid receptor antagonist, AM281, to block these three functional responses were similar, supporting the view that anandamide actions in parotid glands were achieved through a single receptor subtype, the CB1. Binding studies using the selective cannabinoid CB1 receptor antagonist, [3H]SR141716A, indicated the presence of the specific binding site. It may be concluded that in parotid glands the endogenous cannabinoid anandamide, bound to the CB1 cannabinoid receptor subtype, induces cAMP accumulation which in turn leads to amylase release and Na+-K+-ATPase inhibition.

Cholinergic regulation of Na+-K+-ATPase activity in rat parotid gland: changes after castration.

In this study, we investigated the different signalling pathways involved in muscarinic acetylcholine M(3) receptor-dependent modulation of Na(+)-K(+)-ATPase in parotid glands from normal and castrated rats. Carbachol inhibited the enzyme activity in parotid glands from control rats while it stimulated the enzyme activity in castrated rats. The inhibition of Ca(2+) calmodulin by trifluoperazine abolished the inhibitory effect of carbachol in control rats, while the inhibition of protein kinase C by staurosporine stimulated Na(+)-K(+)-ATPase. In castrated rats, trifluoperazine inhibited the carbachol-stimulant effect while staurosporine had no effect. Results indicate that in control glands the activation of a phospholipid-Ca(2+) calmodulin-dependent protein kinase C is responsible for the inhibitory effect of carbachol on Na(+)-K(+)-ATPase activity. In castrated rats, the activation of the enzyme by carbachol is regulated by its Ca(2+) calmodulin-stimulating action, and not by activation of protein kinase C. The activation of the Na(+)-K(+)-ATPase observed in castrated rats resulted in a decrease in carbachol-induced net K(+) efflux and thereby could decrease salivary fluid production.

Castration decreases amylase release associated with muscarinic acetylcholine receptor downregulation in rat parotid gland.

1 The mechanism and receptor subtypes involved in carbachol-stimulated amylase release and its changes after castration were studied in parotid slices from male rats. 2 Carbachol induced both amylase release and inositol phosphate (IP) accumulation in parotid slices from control and castrated rats, but castration induced a decrease of carbachol maximal effect. The effect of castration was reverted by testosterone replacement. 3 The selective M(1) and M(3) muscarinic receptor antagonists, pirenzepine and 4-diphenylacetoxy-N-methylpiperidine methiodide, respectively, inhibited carbachol-stimulated amylase release and IP accumulation in a dose-dependent manner in parotid slices from control and castrated rats. 4 A diminution of binding sites of muscarinic receptor in parotid membrane from castrated rats was observed. Competition binding assays showed that both, M(1) and M(3) muscarinic receptor subtypes are expressed in membranes of parotid glands from control and castrated rats, M(3) being the greater population. 5 These results suggest that amylase release induced by carbachol in parotid slices is mediated by phosphoinositide accumulation. This mechanism appears to be triggered by the activation of M(1) and M(3) muscarinic receptor subtypes. Castration induced a decrease of the maximal effect of carbachol evoked amylase release and IP accumulation followed by a diminution in the number of parotid gland muscarinic acetylcholine receptors.

Influence of castration on isoprenaline-induced amylase release in parotid gland from male rats.

The purpose of this study was to determine the influence of testosterone, the male sex hormone, on beta-adrenergic agonist-induced amylase secretion from rat parotid glands. Isoprenaline (isoproterenol)-induced amylase secretion was measured in vitro from the parotid glands of control and castrated rats with and without testosterone replacement. The isoprenaline-induced amylase release was reduced in parotid glands from castrated rats compared to controls. The reduction of amylase release by isoprenaline in parotid glands of castrated rats, could be reversed by administration of testosterone. Furthermore, beta-adrenergic receptor density and the level of isoprenaline-evoked cAMP in parotid glands from castrated rats was lower compared to intact rats. Using SQ-22536 (an adenylyl cyclase inhibitor), dibutyryl cAMP (a cAMP analogue) and verapamil (a calcium channel blocker), we conclude that the impairment of amylase release from parotid glands after castration was not related to either adenylyl cyclase activity or cAMP accumulation. Amylase release from the parotid glands of castrated rats appears to be mediated by an increase in calcium ion influx.