Comparison of protease and aminopeptidase activities in meconium: A pilot study

Skarżyńska,Ewa; Wilczyńska,Paulina; Kiersztyn,Bartosz; Żytyńska-Daniluk,Joanna; Jakimiuk,Artur; Lisowska-Myjak,Barbara

doi:10.3892/br.2020.1314

Comparison of protease and aminopeptidase activities in meconium: A pilot study

Authors:
- Ewa Skarżyńska
- Paulina Wilczyńska
- Bartosz Kiersztyn
- Joanna Żytyńska-Daniluk
- Artur Jakimiuk
- Barbara Lisowska-Myjak
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Affiliations: Department of Biochemistry and Clinical Chemistry, Medical University of Warsaw, Warsaw 02-097, Poland, Department of Biochemistry, Warsaw University of Life Sciences - SGGW, Warsaw 02-776, Poland, Microbial Ecology and Environmental Biotechnology Department, Institute of Botany, Faculty of Biology, University of Warsaw; Biological and Chemical Research Centre, Warsaw 02-089, Poland, Clinical Department of Obstetrics, Female Diseases and Gynaecological Oncology, Central Clinical Hospital of The Ministry of The Interior, Warsaw 02-507, Poland, Reproductive Health Department, Institute of Mother and Child, Warsaw 01-211, Poland
Published online on: June 9, 2020 https://doi.org/10.3892/br.2020.1314
Article Number: 7

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Abstract

The successive accumulation of proteases and aminopeptidases in meconium are important physiological components of the intrauterine environment in which a fetus develops. The aim of the present study was to assess the changes in the activities of these enzymes in meconium of healthy infants, and to investigate whether there were any statistically significant associations between activity of the enzymes of interest and the mode of delivery. The activities of proteases and aminopeptidases were determined in meconium portions (n=110) using the substrates BODIPY FL casein and L-leucine-7-amido-4-methylcoumarin hydrochloride, respectively. Serial meconium samples (2-5 per neonate) were collected from healthy infants born vaginally (n=14), and by a cesarean section (n=16). Protease activity (104 RFU/h) was lower in the first meconium sample compared with the final sample from the same infant (3.99±2.03 vs. 5.76±2.24, respectively, mean ± standard deviation; P=0.004). Conversely, there was no significant difference in aminopeptidase activity (103 nM/l/h) between consecutive meconium samples (P=0.702). The ratios of the first-meconium sample enzyme activity to the last-meconium sample enzyme activity were lower for proteases compared with aminopeptidases (0.76±0.48 vs. 1.35±1.04, respectively mean ± standard deviation; P=0.014), and sustained in the infants born by a cesarean section (P=0.008). Spearman's correlation coefficient analysis between the first and last meconium samples showed the correlation increased in the infants born vaginally compared with the rest of the infants (proteases, R=0.618 vs. R=0.314; aminopeptidases, R=0.688 vs. R=0.566). Aminopeptidase activity did not exhibit any notable dynamic changes during meconium accumulation in the fetal intestine. In infants born vaginally compared with those born by a cesarean section, the activity of both proteases and aminopeptidases in the first meconium sample showed an improved correlation with the activity of the final meconium sample. This may suggest that in the intrauterine environment, during accumulation of meconium in the digestive tract of the fetus, the activity and/or levels of these enzymes and the substrates they catalyze were more stable in newborns born vaginally compared with infants born by caesarean section.

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1	Pérez-Silva JG, Español Y, Velasco G and Quesada V: The Degradome database: Expanding roles of mammalian proteases in life and disease. Nucleic Acids Res. 44:D351–D355. 2016.PubMed/NCBI View Article : Google Scholar
2	López-Otín C and Bond JS: Proteases: Multifunctional enzymes in life and disease. J Biol Chem. 283:30433–30437. 2008.PubMed/NCBI View Article : Google Scholar
3	Antalis TM, Shea-Donohue T, Vogel SN, Sears C and Fasano A: Mechanisms of disease: Protease functions in intestinal mucosal pathobiology. Nat Clin Pract Gastroenterol Hepatol. 4:393–402. 2007.PubMed/NCBI View Article : Google Scholar
4	Carroll IM, Ringel-Kulka T, Ferrier L, Wu MC, Siddle JP, Bueno L and Ringel Y: Fecal protease activity is associated with compositional alterations in the intestinal microbiota. PLoS One. 8(e78017)2013.PubMed/NCBI View Article : Google Scholar
5	Seshagiri PB, Lalitha HS, Mishra A and Sireesha GV: Embryo-endometrial proteases during early mammalian development. Indian J Exp Biol. 41:756–763. 2003.PubMed/NCBI
6	Mizutani S and Tomoda Y: Effects of placental proteases on maternal and fetal blood pressure in normal pregnancy and preeclampsia. Am J Hypertens. 9:591–597. 1996.PubMed/NCBI View Article : Google Scholar
7	Mizutani S, Wright J and Kobayashi H: A new approach regarding the treatment of preeclampsia and preterm labor. Life Sci. 88:17–23. 2011.PubMed/NCBI View Article : Google Scholar
8	Mizutani S: Physiological roles of placental proteases in feto-placental homeostasis. Nagoya J Med Sci. 61:85–95. 1998.PubMed/NCBI
9	Townsend R, O'Brien P and Khalil A: Current best practice in the management of hypertensive disorders in pregnancy. Integr Blood Press Control. 9:79–94. 2016.PubMed/NCBI View Article : Google Scholar
10	Kattah AG and Garovic VD: The management of hypertension in pregnancy. Adv Chronic Kidney Dis. 20:229–239. 2013.PubMed/NCBI View Article : Google Scholar
11	Sanderink GJ, Artur Y and Siest G: Human aminopeptidases: A review of the literature. J Clin Chem Clin Biochem. 26:795–807. 1988.PubMed/NCBI View Article : Google Scholar
12	Byzia A, Szeffler A, Kalinowski L and Drag M: Activity profiling of aminopeptidases in cell lysates using a fluorogenic substrate library. Biochimie. 122:31–37. 2016.PubMed/NCBI View Article : Google Scholar
13	Park BY and Lee BK: Use of meconium in perinatal epidemiology: Potential benefits and pitfalls. Ann Epidemiol. 24:878–881. 2014.PubMed/NCBI View Article : Google Scholar
14	Shi YC, Guo H, Chen J, Sun G, Ren RR, Guo MZ, Peng LH and Yang YS: Initial meconium microbiome in Chinese neonates delivered naturally or cesarean section. Sci Rep. 8(3255)2018.PubMed/NCBI View Article : Google Scholar
15	Nagpal R, Tsuji H, Takahashi T, Kawashima K, Nagata S, Nomoto K and Yamashiro Y: Sensitive quantitative analysis of the meconium bacterial microbiota in healthy term infants born vaginally or by cesarean section. Front Microbiol. 7(1997)2016.PubMed/NCBI View Article : Google Scholar
16	World Medical Association: World medical association declaration of Helsinki: Ethical principles for medical research involving human subjects. JAMA 310: 2191-2194, 2013.
17	Hoppe HG: Significance of exoenzymatic activities in the ecology of brackish water: Measurements by means of methylumbelliferyl-substrates. Mar Ecol Prog Ser. 11:299–308. 1983.
18	Kiersztyn B, Siuda W and Chróst RJ: Persistence of bacterial proteolytic enzymes in lake ecosystems. FEMS Microbiol Ecol. 80:124–134. 2012.PubMed/NCBI View Article : Google Scholar
19	Chróst RJ: Microbial ectoenzymes in aquatic environments. In: Overbeck J & Chróst RJ, eds. Aquatic microbial ecology: Biochemical and molecular approaches. Springer-Verlag: New York, 47-78, 1990.
20	Johnson KA and Goody RS: The original Michaelis constant: Translation of the 1913 Michaelis-Menten paper. Biochemistry. 50:8264–8269. 2011.PubMed/NCBI View Article : Google Scholar
21	Lisowska-Myjak B, Skarżyńska E, Wojdan K and Nasierowska-Guttmejer A: Protein and peptide profiles in neonatal meconium. J Obstet Gynaecol Res. 45:556–564. 2019.PubMed/NCBI View Article : Google Scholar
22	Skarżyńska E, Kiersztyn B, Wilczyńska P, Jakimiuk A and Lisowska-Myjak B: Total proteolytic activity and concentration of alpha-1 antitrypsin in meconium for assessment of the protease/antiprotease balance. Eur J Obstet Gynecol Reprod Biol. 223:133–138. 2018.PubMed/NCBI View Article : Google Scholar
23	Rawlings ND, Barrett AJ, Thomas PD, Huang X, Bateman A and Finn RD: The MEROPS database of proteolytic enzymes, their substrates and inhibitors in 2017 and a comparison with peptidases in the PANTHER database. Nucleic Acids Res. 46:D624–D632. 2018.PubMed/NCBI View Article : Google Scholar
24	Sugaya N, Takeuchi Y and Kanno T: Increased cytosol aminopeptidase in serum in measles infection. Clin Chem. 34:212–213. 1988.PubMed/NCBI
25	Yamahara N, Nomura S, Suzuki T, Itakura A, Ito M, Okamoto M, Tsujimoto M, Nakazato M and Mizutani S: Placental leucine aminopeptidase/oxytocinase in maternal serum and placenta during normal pregnancy. Life Sci. 66:1401–1410. 2000.PubMed/NCBI View Article : Google Scholar
26	Kobayashi H, Nomura S, Mitsui T, Ito T, Kuno N, Ohno Y, Kadomatsu K, Muramatsu K, Nagasaka T and Mizutani S: Tissue distribution of placental leucine aminopeptidase/oxytocinase during mouse pregnancy. J Histochem Cytochem. 52:113–121. 2004.PubMed/NCBI View Article : Google Scholar
27	Matsumoto H and Mori T: Changes in cystine aminopeptidase (oxytocinase) activity in mouse serum, placenta, uterus and liver during pregnancy or after steroid hormone treatments. Zoolog Sci. 15:111–115. 1998.PubMed/NCBI View Article : Google Scholar
28	Grzywa R, Oleksyszyn J, Salvesen GS and Drag M: Identification of very potent inhibitor of human aminopeptidase N (CD13). Bioorg Med Chem Lett. 20:2497–2499. 2010.PubMed/NCBI View Article : Google Scholar
29	Gard PR, Fidalgo S, Lotter I, Richardson C, Farina N, Rusted J and Tabet N: Changes of renin-angiotensin system-related aminopeptidases in early stage Alzheimer's disease. Exp Gerontol. 89:1–7. 2017.PubMed/NCBI View Article : Google Scholar
30	Gonzales T and Robert-Baudouy J: Bacterial aminopeptidases: Properties and functions. FEMS Microbiol Rev. 18:319–344. 1996.PubMed/NCBI View Article : Google Scholar
31	Neu J and Rushing J: Cesarean versus vaginal delivery: Long-term infant outcomes and the hygiene hypothesis. Clin Perinatol. 38:321–331. 2011.PubMed/NCBI View Article : Google Scholar
32	Arboleya S, Suárez M, Fernández N, Mantecón L, Solís G, Gueimonde M, de Los Reyes and Gavilán CG: C-section and the neonatal gut microbiome acquisition: Consequences for future health. Ann Nutr Metab. 73 (Suppl 3):17–23. 2018.PubMed/NCBI View Article : Google Scholar