Top

Lung

Published in:

01-04-2019 | Foreign Body Aspiration | ACUTE LUNG INJURY

Degradation of Lung Protective Angiotensin Converting Enzyme-2 by Meconium in Human Alveolar Epithelial Cells: A Potential Pathogenic Mechanism in Meconium Aspiration Syndrome

Authors: Chintan K. Gandhi, Romel Holmes, Ira H. Gewolb, Bruce D. Uhal

Published in: Lung | Issue 2/2019

Abstract

Background

Pancreatic digestive enzymes present in meconium might be responsible for meconium-induced lung injury. The local Renin Angiotensin System plays an important role in lung injury and inflammation. Particularly, angiotensin converting enzyme-2 (ACE-2) has been identified as a protective lung enzyme against the insult. ACE-2 converts pro-apoptotic Angiotensin II to anti-apoptotic Angiotensin 1–7. However, the effect of meconium on ACE-2 has never been studied before.

Objective

To study the effect of meconium on ACE-2, and whether inhibition of proteolytic enzymes present in the meconium reverses its effects on ACE-2.

Methods

Alveolar epithelial A549 cells were exposed to F-12 medium, 2.5% meconium, meconium + a protease inhibitor cocktail (PIc) and PIc alone for 16 h. At the end of incubation, apoptosis was measured with a nuclear fragmentation assay and cell lysates were collected for ACE-2 immunoblotting and enzyme activity.

Results

Meconium caused a fourfold increase in apoptotic nuclei (p < 0.001). The pro-apoptotic effect of meconium can be reversed by PIc. Meconium reduced ACE-2 enzyme activity by cleaving ACE-2 into a fragment detected at ~ 37 kDa by immunoblot. PIc prevented the degradation of ACE-2 and restored 50% of ACE-2 activity (p < 0.05).

Conclusion

These data suggest that meconium causes degradation of lung protective ACE-2 by proteolytic enzymes present in meconium, since the effects of meconium can be reversed by PIc.

Gandhi CK (2018) Management of meconium-stained newborns in the delivery room. Neonatal Netw NN 37(3):141–148. https://doi.org/10.1891/0730-0832.37.3.141 CrossRefPubMed

Tyler DC, Murphy J, Cheney FW (1978) Mechanical and chemical damage to lung tissue caused by meconium aspiration. Pediatrics 62(4):454–459PubMed

Zagariya A, Bhat R, Chari G, Uhal B, Navale S, Vidyasagar D (2005) Apoptosis of airway epithelial cells in response to meconium. Life Sci 76(16):1849–1858. https://doi.org/10.1016/j.lfs.2004.10.033 CrossRefPubMed

Gandhi C, Uhal BD (2016) Roles of the angiotensin system in neonatal lung injury and disease. JSM Atheroscler 1(3):1014PubMedPubMedCentral

Sparks MA, Crowley SD, Gurley SB, Mirotsou M, Coffman TM (2014) Classical renin-angiotensin system in kidney physiology. Compr Physiol 4(3):1201–1228. https://doi.org/10.1002/cphy.c130040 CrossRefPubMedPubMedCentral

Yoshiji H, Kuriyama S, Yoshii J, Ikenaka Y, Noguchi R, Nakatani T, Tsujinoue H, Fukui H (2001) Angiotensin-II type 1 receptor interaction is a major regulator for liver fibrosis development in rats. Hepatology 34(4 Pt 1):745–750. https://doi.org/10.1053/jhep.2001.28231 CrossRefPubMed

Paul M, Poyan Mehr A, Kreutz R (2006) Physiology of local renin-angiotensin systems. Physiol Rev 86(3):747–803. https://doi.org/10.1152/physrev.00036.2005 CrossRefPubMed

Chen LN, Yang XH, Nissen DH, Chen YY, Wang LJ, Wang JH, Gao JL, Zhang LY (2013) Dysregulated renin-angiotensin system contributes to acute lung injury caused by hind-limb ischemia-reperfusion in mice. Shock (Augusta GA) 40(5):420–429. https://doi.org/10.1097/SHK.0b013e3182a6953e CrossRef

Liu L, Qiu HB, Yang Y, Wang L, Ding HM, Li HP (2009) Losartan, an antagonist of AT1 receptor for angiotensin II, attenuates lipopolysaccharide-induced acute lung injury in rat. Arch Biochem Biophys 481(1):131–136. https://doi.org/10.1016/j.abb.2008.09.019 CrossRefPubMed

10.

Marshall RP (2003) The pulmonary renin-angiotensin system. Curr Pharm Des 9(9):715–722CrossRefPubMed

11.

Zhang H, Sun GY (2005) LPS induces permeability injury in lung microvascular endothelium via AT(1) receptor. Arch Biochem Biophys 441(1):75–83. https://doi.org/10.1016/j.abb.2005.06.022 CrossRefPubMed

12.

Wang R, Alam G, Zagariya A, Gidea C, Pinillos H, Lalude O, Choudhary G, Oezatalay D, Uhal BD (2000) Apoptosis of lung epithelial cells in response to TNF-alpha requires angiotensin II generation de novo. J Cell Physiol 185 (2):253–259. https://doi.org/10.1002/1097-4652(200011)185:2%3C253::Aid-jcp10%3E3.0.Co;2-%23 CrossRefPubMed

13.

Li X, Molina-Molina M, Abdul-Hafez A, Uhal V, Xaubet A, Uhal BD (2008) Angiotensin converting enzyme-2 is protective but downregulated in human and experimental lung fibrosis. Am J Physiol Lung Cell Mol Physiol 295(1):L178–L185. https://doi.org/10.1152/ajplung.00009.2008 CrossRefPubMedPubMedCentral

14.

Antonowicz I, Shwachman H (1979) Meconium in health and in disease. Adv Pediatr 26:275–310PubMed

15.

Ivanov VA, Gewolb IH, Uhal BD (2010) A new look at the pathogenesis of the meconium aspiration syndrome: a role for fetal pancreatic proteolytic enzymes in epithelial cell detachment. Pediatr Res 68(3):221–224. https://doi.org/10.1203/00006450-201011001-00431 CrossRefPubMed

16.

Uhal BD, Dang M, Dang V, Llatos R, Cano E, Abdul-Hafez A, Markey J, Piasecki CC, Molina-Molina M (2013) Cell cycle dependence of ACE-2 explains downregulation in idiopathic pulmonary fibrosis. Eur Respir J 42(1):198–210. https://doi.org/10.1183/09031936.00015612 CrossRefPubMed

17.

Xiao F, Burns KD (2017) Measurement of angiotensin converting enzyme 2 activity in biological fluid (ACE2). Methods Mol Biol (Clifton NJ) 1527:101–115. https://doi.org/10.1007/978-1-4939-6625-7_8 CrossRef

18.

Li X, Zhang H, Soledad-Conrad V, Zhuang J, Uhal BD (2003) Bleomycin-induced apoptosis of alveolar epithelial cells requires angiotensin synthesis de novo. Am J Physiol Lung Cell Mol Physiol 284(3):L501–L507. https://doi.org/10.1152/ajplung.00273.2002 CrossRefPubMed

19.

Uhal BD, Joshi I, Hughes WF, Ramos C, Pardo A, Selman M (1998) Alveolar epithelial cell death adjacent to underlying myofibroblasts in advanced fibrotic human lung. Am J Physiol 275(6 Pt 1):L1192–L1199PubMed

20.

Wosten-van Asperen RM, Lutter R, Specht PA, Moll GN, van Woensel JB, van der Loos CM, van Goor H, Kamilic J, Florquin S, Bos AP (2011) Acute respiratory distress syndrome leads to reduced ratio of ACE/ACE2 activities and is prevented by angiotensin-(1–7) or an angiotensin II receptor antagonist. J Pathol 225(4):618–627. https://doi.org/10.1002/path.2987 CrossRefPubMed

21.

Garg M, Angus PW, Burrell LM, Herath C, Gibson PR, Lubel JS (2012) Review article: the pathophysiological roles of the renin-angiotensin system in the gastrointestinal tract. Aliment Pharmacol Ther 35(4):414–428. https://doi.org/10.1111/j.1365-2036.2011.04971.x CrossRefPubMedPubMedCentral

22.

Mullinger M, Palasi M (1966) Tryptic and chymotryptic activity of stools of newborn infants. Pediatrics 38(4):657–659PubMed

23.

Wyllie AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284(5756):555–556CrossRefPubMed

24.

Riccardi C, Nicoletti I (2006) Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1(3):1458–1461. https://doi.org/10.1038/nprot.2006.238 CrossRefPubMed

25.

Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139(2):271–279CrossRefPubMed

26.

Dini L, Coppola S, Ruzittu MT, Ghibelli L (1996) Multiple pathways for apoptotic nuclear fragmentation. Exp Cell Res 223(2):340–347. https://doi.org/10.1006/excr.1996.0089 CrossRefPubMed

27.

Oberhammer F, Fritsch G, Schmied M, Pavelka M, Printz D, Purchio T, Lassmann H, Schulte-Hermann R (1993) Condensation of the chromatin at the membrane of an apoptotic nucleus is not associated with activation of an endonuclease. J Cell Sci 104(Pt 2):317–326PubMed

28.

Ghibelli L, Maresca V, Coppola S, Gualandi G (1995) Protease inhibitors block apoptosis at intermediate stages: a compared analysis of DNA fragmentation and apoptotic nuclear morphology. FEBS Lett 377(1):9–14CrossRefPubMed

29.

Gopallawa I, Uhal BD (2016) Angiotensin-(1–7)/mas inhibits apoptosis in alveolar epithelial cells through upregulation of MAP kinase phosphatase-2. Am J Physiol Lung Cell Mol Physiol 310(3):L240–L248. https://doi.org/10.1152/ajplung.00187.2015 CrossRefPubMed

30.

Zagariya A, Bhat R, Uhal B, Navale S, Freidine M, Vidyasagar D (2000) Cell death and lung cell histology in meconium aspirated newborn rabbit lung. Eur J Pediatr 159(11):819–826CrossRefPubMed

31.

Rosenfeld CR, Zagariya AM, Liu XT, Willis BC, Fluharty S, Vidyasagar D (2008) Meconium increases type 1 angiotensin II receptor expression and alveolar cell death. Pediatr Res 63(3):251–256. https://doi.org/10.1203/PDR.0b013e318163a2b8 CrossRefPubMed

32.

Huang L, Sexton DJ, Skogerson K, Devlin M, Smith R, Sanyal I, Parry T, Kent R, Enright J, Wu QL, Conley G, DeOliveira D, Morganelli L, Ducar M, Wescott CR, Ladner RC (2003) Novel peptide inhibitors of angiotensin-converting enzyme 2. J Biol Chem 278(18):15532–15540. https://doi.org/10.1074/jbc.M212934200 CrossRefPubMed

33.

Kuba K, Imai Y, Rao S, Gao H, Guo F, Guan B, Huan Y, Yang P, Zhang Y, Deng W, Bao L, Zhang B, Liu G, Wang Z, Chappell M, Liu Y, Zheng D, Leibbrandt A, Wada T, Slutsky AS, Liu D, Qin C, Jiang C, Penninger JM (2005) A crucial role of angiotensin converting enzyme 2 (ACE2) in SARS coronavirus-induced lung injury. Nat Med 11(8):875–879. https://doi.org/10.1038/nm1267 CrossRefPubMedPubMedCentral

34.

Imai Y, Kuba K, Rao S, Huan Y, Guo F, Guan B, Yang P, Sarao R, Wada T, Leong-Poi H, Crackower MA, Fukamizu A, Hui CC, Hein L, Uhlig S, Slutsky AS, Jiang C, Penninger JM (2005) Angiotensin-converting enzyme 2 protects from severe acute lung failure. Nature 436(7047):112–116. https://doi.org/10.1038/nature03712 CrossRefPubMedPubMedCentral

35.

Wiener RS, Cao YX, Hinds A, Ramirez MI, Williams MC (2007) Angiotensin converting enzyme 2 is primarily epithelial and is developmentally regulated in the mouse lung. J Cell Biochem 101(5):1278–1291. https://doi.org/10.1002/jcb.21248 CrossRefPubMedPubMedCentral

36.

Oarhe CI, Dang V, Dang M, Nguyen H, Gopallawa I, Gewolb IH, Uhal BD (2015) Hyperoxia downregulates angiotensin-converting enzyme-2 in human fetal lung fibroblasts. Pediatr Res 77(5):656–662. https://doi.org/10.1038/pr.2015.27 CrossRefPubMedPubMedCentral

37.

Mohamed TL, Nguyen HT, Abdul-Hafez A, Dang VX, Dang MT, Gewolb IH, Uhal BD (2016) Prior hypoxia prevents downregulation of ACE-2 by hyperoxia in fetal human lung fibroblasts. Exp Lung Res 42(3):121–130. https://doi.org/10.3109/01902148.2016.1157712 CrossRefPubMedPubMedCentral

38.

Khan A, Benthin C, Zeno B, Albertson TE, Boyd J, Christie JD, Hall R, Poirier G, Ronco JJ, Tidswell M, Hardes K, Powley WM, Wright TJ, Siederer SK, Fairman DA, Lipson DA, Bayliffe AI, Lazaar AL (2017) A pilot clinical trial of recombinant human angiotensin-converting enzyme 2 in acute respiratory distress syndrome. Critic Care (Lond Engl) 21(1):234. https://doi.org/10.1186/s13054-017-1823-x CrossRef

Title: Degradation of Lung Protective Angiotensin Converting Enzyme-2 by Meconium in Human Alveolar Epithelial Cells: A Potential Pathogenic Mechanism in Meconium Aspiration Syndrome
Authors: Chintan K. Gandhi
Romel Holmes
Ira H. Gewolb
Bruce D. Uhal
Publication date: 01-04-2019
Publisher: Springer US
Keywords: Foreign Body Aspiration
Foreign Body Aspiration
Aspiration
Published in: Lung / Issue 2/2019
Print ISSN: 0341-2040
Electronic ISSN: 1432-1750
DOI: https://doi.org/10.1007/s00408-019-00201-y

ACC 2024 Congress Coverage

Heart Failure Video

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Degradation of Lung Protective Angiotensin Converting Enzyme-2 by Meconium in Human Alveolar Epithelial Cells: A Potential Pathogenic Mechanism in Meconium Aspiration Syndrome

Abstract

Background

Objective

Methods

Results

Conclusion

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Objective

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 2/2019

Using a Dedicated Interventional Pulmonology Practice Decreases Wait Time Before Treatment Initiation for New Lung Cancer Diagnoses

The Role of the Cerebellum in Control of Swallow: Evidence of Inspiratory Activity During Swallow

Nanogold Assay Improves Accuracy of Conventional TB Diagnostics

The Effect of Breathing Retraining Using Metronome-Based Acoustic Feedback on Exercise Endurance in COPD: A Randomized Trial

Fractional Exhaled Nitric Oxide Measurements and Screening of Obstructive Sleep Apnea in a Sleep-Laboratory Setting: A Cross-Sectional Study

The Burden of Sarcoidosis Symptoms from a Patient Perspective

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page