42 J Meikai Dent Med 371, 42 49, 2008 A ACgA CgACorAml CgA, Cor, Aml CgA CgA, Aml CgA Aml Cor CgA CgA CgA CgA CgA CgA Salivary Chromogranin A as Index of Psychosomatic Stress Response Takeya TSUNODA Division of Pediatric Dentistry, Department of Human Development & Fostering, Meikai University Scool of Dentistry Director : Prof Shigeru WATANABE Abstract : It is reported that chromogranin ACgAin the saliva is useful as a psychological stress marker. The purpose of this study is to examine the effect of salivary flow rates on the parotid salivary concentrations of CgA, cortisolcorand amylaseamland also is to examine their changes before and after chewing stimulation by wax, and the change of CgA volume before and after the infiltration anesthesia action on the whole saliva. Results obtained were as follows ; CgA and Aml concentrations of the stimulated parotid saliva were significantly decreased in comparison with the unstimulated saliva. There were no significant effects of the saliva sampling time on any of the measurements. The mean values of CgA after chewing wax for each subject tended to decrease as compared to those from before chewing wax. The Aml values showed an increasing tendency, while the Cor values did not show any definite tendency. There was significant correlation between salivary flow rate and CgA concentration in the saliva. CgA volume in the saliva after wax chewing significantly increased in comparison with that before wax chewing. CgA volume in the saliva after anesthesia action was significantly increased from that of before anesthesia action. In conclusion, it is necessary to evaluate the CgA volume in saliva when we use it as an index for psychological stress. The CgA volume in saliva is expected to serve as an index for stress evaluation in dental treatment. Key words : salivary CgA, salivary flow rate, stress evaluation, infiltration anesthesia
A 43 A 0503 1, 2 2 4 5, 6 7 2 8, 9 10, 11 A CgA 1 CgA, Cor, Aml 1 8 4 4 2138 2 Lashley 0.5 ml 26 mmol/l 0.5 ml 30 3000 g 80 1 1 10 : 003 CgA, Cor, Aml Fig 1 3 Model 2001 MP 2, Sartorius, Goettingen, Germany CgA 11, 12 CgA 12, 13 CgA 14 21 CgA 1 CgACorAml 2 5 CgA, Cor, Aml 3 CgA 350-0283 1-1 Fig 1 Modified Carlson-Crittenden collector for parotid saliva.
44 37, 2008 Omniscribe, Industrial Scientific, Houston, TX, USA 0.01 ml 5cm 1 CgA, Cor, Aml 4 CgACgA CgA EIA YK 070 10 mm phosphate bufferph 7.430 mm NaCl, 5 mm EDTA, 0.1 BSA CgA CgA0.25 mg/well CgA IgG streptavidinhorseradish peroxidase 100 µl 2 o-phenylene diamine 0.015 H2O2, 0.1 M phosphate-citrate bufferph 7.4100 µl 30 100 µl Labsystem Multiskan Bichromatic, Helsinki, Finland 490 nm CgA 7 CgA CorCor Cortisol EIA No 900-071, Assay Designs, Ann Arbor, MI, USA Assay Buffer 10 Cor alkaline phosphatase Cor Cor IgG 2 p-nitrophenylphosphate 200 µl 1 50 µl 405 nm Cor 7 Cor Aml phosphate-buffered saline 1/200 salivary α -amylase assay kitstate Collage, PA, USA8 µl 37 2-chloro-p-nitrophenol linked with maltotriose 320 µl 37 1 3 405 nm α -Amylase U/ml Absorbance/minTVDF α -Amylase MMASVLP U/ml Absorbance/min1 TVTotal assay volume 0.328 ml DFDiluent factor 200 MMA12.9 SVSample volume 0.008 ml LPLight path 0.97 2 CgA, Cor, Aml 1 6 3 3 2735 2 1 5 5 g 5 5 1 3 1000, 1400, 17 003 CgA, Cor, Aml 3 CgA Cor Aml 1 3 CgA 1 20 13 7 2334 2 2 5 9:30
A 45 2 5 CgA 3 CgA 1 4 1 CgA 1 3 Student s t-test 2 Spearman p 0.05 CgA, Cor, Aml Table 1 0.05 0.02 ml /min 0.180.10 ml /min CgA 13.111.8 pmol/ml 3.7 1.0 pmol/ml Aml 177.971.9 U/ml 64.8 26.8 U/ml Cor 2 Fig 2 1 6 2 11 Table 2 CgA, Cor, Aml 1 Table 1 Parotid salivary flow rate and concentrations of CgA, Cor and Aml in parotid saliva before and after the stimulation. Before After Parotid salivary flow rateml /min CgApmol/ml Corpg/ml AmlU/ml 0.050.02 13.111.8 583.9228.5 177.971.9 0.180.10** 3.71.0* 600.9341.1 64.826.8** CgA : Chromogranin A, Cor : Cortisol, Aml : Amylase. *P 0.05 vs before the stimulation, **P 0.01, n9 Fig 2 The whole salivary flow rate before and after chewing wax. Table 2 Concentrations of CgA, Cor and Aml in whole saliva before and after chewing wax. CgApmol/ml Corpg/ml AmlU/ml before after before after before after 3.12.2 23.79.2 11.36.0 9.72.7 9.50.7 12.23.2 2.0*1.7 11.0**9.4 5.5*2.5 1.7**1.0 5.8**1.7 7.0**3.0 1303.1 321.3 1226.9 598.8 2211.01133.7 1002.7*620.5 1117.7 445.3 2054.11067.3 731.0 438.0 819.0*417.5 616.0 94.7 881.3** 35.1 2914.81337.9 808.3**332.8 10.5 6.8 5.4 3.4 40.121.4 3.4 2.1 5.2 1.0 28.2 4.6 47.6**13.9 9.1** 1.8 46.5 24.4 6.7 4.3 30.2**16.9 45.4**14.7 CgA : Chromogranin A, Cor : Cortisol, Aml : Amylase. SD, *P 0.05, **P 0.01, n9
46 37, 2008 Table 3 Correlation between salivary flow rate and concentration of Chromogranin A in saliva. Table 5 Mean values of salivary flow rate, CgA concentration and CgA volume at rest position. r 0.92 0.76 0.78 0.87 0.74 0.69 p value p0.01 p0.01 p0.01 p0.01 p0.01 p0.01 r: correlation coefficient, n9 Table 4 Mean concentrations of Chromogranin A before and after chewing wax. Before pmol/min 0.720.43 3.721.40 2.821.09 3.251.27 2.800.25 2.300.88 After pmol/min 2.271.40* 16.629.92* 10.496.56* 9.093.52* 8.931.60* 11.946.04* *P 0.01MeanSD, n9 Sub 7 Sub 8 Sub 9 0 1 2 3 4 5 6 7 8 9 0 Salivary flow rate ml /min 0.3 0.09 0.400.11 0.250.07 0.400.13 0.480.08 0.190.06 0.220.06 0.430.10 0.370.11 0.120.05 0.330.10 0.440.13 0.380.12 0.360.15 0.290.08 0.320.15 0.250.09 0.280.11 0.340.13 0.300.09 CgA : Chromogranin A. n5. CgA concentration pmol/ml 3.20.8 4.91.2 7.31.0 5.01.4 4.11.1 3.10.8 3.21.0 2.80.8 1.90.9 2.80.7 2.90.9 1.60.7 3.10.9 3.01.7 4.91.5 3.61.3 2.81.1 3.21.9 2.81.2 2.51.3 CgA Volume pmol/min 0.960.08 1.960.15 1.820.05 2.000.11 1.960.06 0.580.10 0.700.13 1.200.09 0.700.09 0.330.15 0.950.12 0.700.11 1.170.12 1.080.14 1.420.07 1.150.13 0.700.10 0.890.09 0.950.11 0.750.10 CgA Cor 3 1 Aml 6 4 Table 3 CgA 0.69 0.92 CgA CgA 1 CgA Table 4 CgA p0.01 3 CgA 1 CgA Table 5 CgA SD Table 6 CgA Table 6 Salivary frow rate, CgA concentration and CgA volume just after anesthesia action. Sub 7 Sub 8 Sub 9 0 1 2 3 4 5 6 7 8 9 0 Salivary flow rate ml /min 0.34 0.45 0.28 0.24 0.21 0.22 0.25 0.48 0.3 0.35 0.37 0.45 0.34 0.12 0.32 0.37 0.49 0.31 0.37 0.33 CgA : Chromogranin A. CgA concentration pmol/ml 6.3 5.8 9.6 5.3 4.8 9.7 4.8 3.9 3.3 1.8 3.6 2.9 4.1 3.8 6.1 4.4 2.9 3.6 3.1 2.8 CgA Volume pmol/min 2.14 2.61 2.68 1.27 1.00 2.13 1.2 1.87 0.99 0.63 1.33 1.3 1.39 0.45 1.95 1.62 1.42 1.11 1.14 0.92
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