RESEARCH PAPER
Caspase 3 activity in isolated fetal rat lung fibroblasts and rat periodontal ligament fibroblasts: cigarette smoke induced alterations
| 1 | Departments of Oral Biology, University of Manitoba and Manitoba Institute for Child Health, Winnipeg, Canada |
| 2 | Human Anatomy and Cell Science, University of Manitoba and Manitoba Institute for Child Health, Winnipeg, Canada |
| 3 | The National Research Council Biodiagnostics Institute, University of Manitoba and Manitoba Institute for Child Health, Winnipeg, Canada |
| 4 | The Biology of Breathing Group, University of Manitoba and Manitoba Institute for Child Health, Winnipeg, Canada |
CORRESPONDING AUTHOR
James E Scott
Departments of Oral Biology, University of Manitoba and Manitoba Institute for Child Health, Winnipeg, Canada
Departments of Oral Biology, University of Manitoba and Manitoba Institute for Child Health, Winnipeg, Canada
Publication date: 2013-12-06
Tobacco Induced Diseases 2013;11(December):25
KEYWORDS
cigarette smoke extractfetal rat lung fibroblastsproteasecaspase-3lung developmentdevelopmental toxicityperiodontal ligament fibroblastperiodontitis
ABSTRACT
Background:
Cigarette smoking is the leading cause of preventable death and has been implicated in pathogenesis of pulmonary, oral and systemic diseases. Smoking during pregnancy is a risk factor for the developing fetus and may be a major cause of infant mortality. Moreover, the oral cavity, and all cells within are the first to be exposed to cigarette smoke and may be a possible source for the spread of toxins to other organs of the body. Fibroblasts in general are morphologically heterogeneous connective tissue cells with diverse functions. Apoptosis or programmed cell death is a crucial process during embryogenesis and for the maintenance of homeostasis throughout life. Deregulation of apoptosis has been implicated in abnormal lung development in the fetus and disease progression in adults. Caspases are proteases which belong to the family of cysteine aspartic acid proteases and are key components for downstream amplification of intracellular apoptotic signals. Of 14 known caspases, caspase-3 is the key executioner of apoptosis. In the present study we explored the hypothesis that cigarette smoke (CS) extract activates caspase-3 in two types of fibroblasts, both of which would be exposed directly to cigarette smoke, isolated fetal rat lung fibroblasts and adult rat periodontal ligament (PDL) fibroblasts.
Methods:
Isolated fetal rat lung fibroblasts and adult PDLs were used. Cells were exposed to different concentrations of CS for 60 min. Caspase-3 activity and its inhibition by Z-VAD-fmk were measured by caspase-3 fluorometric assay. The effect of CSE on cellular viability was measured using the MTT formazan assay. Caspase-3 expression was detected by western blot analysis and cellular localization of caspase-3 was determined by immunofluorescence using fluorescence microscopy.
Results:
It was observed in fetal rat lung fibroblast cells that CSE extract significantly (p<0.05) increased caspase-3 activity and decrease cell proliferation. However, no significant changes in activity or viability were observed in PDLs.
Conclusions:
This indicates CS activates caspase-3 the key regulatory point in apoptosis in fetal rat lung fibroblast cells suggesting that smoking during pregnancy may alter the developmental program of fetal lung, jeopardizing the establishment of critical cellular mechanisms necessary to expedite pulmonary maturation at birth.of critical cellular mechanisms necessary to expedite pulmonary maturation at birth.
Cigarette smoking is the leading cause of preventable death and has been implicated in pathogenesis of pulmonary, oral and systemic diseases. Smoking during pregnancy is a risk factor for the developing fetus and may be a major cause of infant mortality. Moreover, the oral cavity, and all cells within are the first to be exposed to cigarette smoke and may be a possible source for the spread of toxins to other organs of the body. Fibroblasts in general are morphologically heterogeneous connective tissue cells with diverse functions. Apoptosis or programmed cell death is a crucial process during embryogenesis and for the maintenance of homeostasis throughout life. Deregulation of apoptosis has been implicated in abnormal lung development in the fetus and disease progression in adults. Caspases are proteases which belong to the family of cysteine aspartic acid proteases and are key components for downstream amplification of intracellular apoptotic signals. Of 14 known caspases, caspase-3 is the key executioner of apoptosis. In the present study we explored the hypothesis that cigarette smoke (CS) extract activates caspase-3 in two types of fibroblasts, both of which would be exposed directly to cigarette smoke, isolated fetal rat lung fibroblasts and adult rat periodontal ligament (PDL) fibroblasts.
Methods:
Isolated fetal rat lung fibroblasts and adult PDLs were used. Cells were exposed to different concentrations of CS for 60 min. Caspase-3 activity and its inhibition by Z-VAD-fmk were measured by caspase-3 fluorometric assay. The effect of CSE on cellular viability was measured using the MTT formazan assay. Caspase-3 expression was detected by western blot analysis and cellular localization of caspase-3 was determined by immunofluorescence using fluorescence microscopy.
Results:
It was observed in fetal rat lung fibroblast cells that CSE extract significantly (p<0.05) increased caspase-3 activity and decrease cell proliferation. However, no significant changes in activity or viability were observed in PDLs.
Conclusions:
This indicates CS activates caspase-3 the key regulatory point in apoptosis in fetal rat lung fibroblast cells suggesting that smoking during pregnancy may alter the developmental program of fetal lung, jeopardizing the establishment of critical cellular mechanisms necessary to expedite pulmonary maturation at birth.of critical cellular mechanisms necessary to expedite pulmonary maturation at birth.
REFERENCES (45)
1.
Roquer JM, Figueras J, Botet F, Jimenez R: Influence on fetal growth of exposure to tobacco smoke during pregnancy. Acta Paediatr. 1995, 84 (2): 118-121. 10.1111/j.1651-2227.1995.tb13592.x.
2.
DiFranza JR, Aligne CA, Weitzman M: Prenatal and postnatal environmental tobacco smoke exposure and children’s health. Pediatrics. 2004, 113 (4 Suppl): 1007-1015.
3.
Chang YC, Huang FM, Tai KW, Yang LC, Chou MY: Mechanisms of cytotoxicity of nicotine in human periodontal ligament fibroblast cultures in vitro. J Periodontal Res. 2002, 37 (4): 279-285. 10.1034/j.1600-0765.2002.01612.x.
4.
Spooner BS, Wessells NK: Mammalian lung development: interactions in primordium formation and bronchial morphogenesis. J Exp Zool. 1970, 175 (4): 445-454. 10.1002/jez.1401750404.
5.
Nishino H, Nemoto N, Lu W, Sakurai I: Significance of apoptosis in morphogenesis of human lung development: light microscopic observation using in situ DNA end-labeling and ultrastructural study. Med Electron Microsc. 1999, 32 (1): 57-61. 10.1007/s007950050009.
6.
Haanen C, Vermes I: Apoptosis: programmed cell death in fetal development. Eur J Obstet Gynecol Reprod Biol. 1996, 64 (1): 129-133. 10.1016/0301-2115(95)02261-9.
7.
Naik AS, Kallapur SG, SG Bachurski CJ, Jobe AH, AH Michna J, Kramer BW, Ikegami M: Effects of ventilation with different positive end-expiratory pressures on cytokine expression in the preterm lamb lung. Am J Respir Crit Care Med. 2001, 164 (3): 494-498. 10.1164/ajrccm.164.3.2010127.
8.
Wright JR: Pulmonary surfactant: a front line of lung host defense. J Clin Invest. 2003, 111 (10): 1453-1455.
9.
Joshi S, Kotecha S: Lung growth and development. Early Hum Dev. 2007, 83 (12): 789-794. 10.1016/j.earlhumdev.2007.09.007.
10.
Habek D, Habek JC, Ivanisevic M, Djelmis J: Fetal tobacco syndrome and perinatal outcome. Fetal Diagn Ther. 2002, 17 (6): 367-371. 10.1159/000065387.
11.
Habek D, Habek JC, Jugovic D, Salihagic A: Intrauterine hypoxia and sudden infant death syndrome. Acta Med Croatica. 2002, 56 (3): 109-118.
12.
Ishii T, Matsuse T, Igarashi H, Masuda M, Teramoto S, Ouchi Y: Tobacco smoke reduces viability in human lung fibroblasts: protective effect of glutathione S-transferase P1. Am J Physiol Lung Cell Mol Physiol. 2001, 280 (6): L1189-L1195.
13.
Kosmider B, Messier EM, Chu HW, Mason RJ: Human alveolar epithelial cell injury induced by cigarette smoke. PLoS One. 2011, 6 (12): e26059-10.1371/journal.pone.0026059.
14.
Krall EA, EA Garvey AJ, Garcia RI: Alveolar bone loss and tooth loss in male cigar and pipe smokers. J Am Dent Assoc. 1999, 130 (1): 57-64.
15.
Nicholson DW, Thornberry NA: Caspases: killer proteases. Trends Biochem Sci. 1997, 22 (8): 299-306. 10.1016/S0968-0004(97)01085-2.
16.
Fadeel B, Orrenius S, Zhivotovsky B: The most unkindest cut of all: on the multiple roles of mammalian caspases. Leukemia. 2000, 14 (8): 1514-1525. 10.1038/sj.leu.2401871.
17.
Degterev A, Boyce M, Yuan J: A decade of caspases. Oncogene. 2003, 22 (53): 8543-8567. 10.1038/sj.onc.1207107.
18.
Green D, Kroemer G: The central executioners of apoptosis: caspases or mitochondria?. Trends Cell Biol. 1998, 8 (7): 267-271. 10.1016/S0962-8924(98)01273-2.
19.
Thornberry NA, Lazebnik Y: Caspases: enemies within. Science. 1998, 281 (5381): 1312-1316.
20.
Tyas L, Brophy VA, Pope A, Rivett AJ, Tavare JM: Rapid caspase-3 activation during apoptosis revealed using fluorescence-resonance energy transfer. EMBO Rep. 2000, 1 (3): 266-270. 10.1093/embo-reports/kvd050.
21.
Giansanti V, Torriglia A, Scovassi AI: Conversation between apoptosis and autophagy: “Is it your turn or mine?”. Apoptosis. 2011, 16: 321-333. 10.1007/s10495-011-0589-x.
22.
Janoff A, Carp H: Possible mechanisms of emphysema in smokers: cigarette smoke condensate suppresses protease inhibition in vitro. Am Rev Respir Dis. 1977, 116 (1): 65-72.
23.
Tabassian AR, Nylen ES, Giron AE, Snider RH, Cassidy MM, Becker KL: Evidence for cigarette smoke-induced calcitonin secretion from lungs of man and hamster. Life Sci. 1988, 42 (23): 2323-2329. 10.1016/0024-3205(88)90185-3.
24.
Hastings C, Rand T, Bergen HT, Thliveris JA, Shaw AR, Lombaert GA, Mantsch HH, Giles BL, Dakshinamurti S, Scott JE: Stachybotrys chartarum alters surfactant-related phospholipid synthesis and CTP:cholinephosphate cytidylyltransferase activity in isolated fetal rat type II cells. Toxicol Sci. 2005, 84 (1): 186-194. 10.1093/toxsci/kfi045.
25.
Lekic PC, Nayak BN, Al-Sanea R, Tenenbaum H, Ganss B, McCulloch C: Cell transplantation in wounded mixed connective tissues. Anat Rec A Discov Mol Cell Evol Biol. 2005, 287 (2): 1256-1263.
26.
Harrington HA, Ho KL, Ghosh S, Tung KC: Construction and analysis of a modular model of caspase activation in apoptosis. Theor Biol Med Model. 2008, 5: 26-10.1186/1742-4682-5-26.
27.
Ott L: An Introduction to Statistical Methods and Data Analysis. 1977, North Scituate: Duxbury Press.
28.
Kilic M, Schafer R, Hoppe J, Kagerhuber U: Formation of noncanonical high molecular weight caspase-3 and −6 complexes and activation of caspase-12 during serum starvation induced apoptosis in AKR-2B mouse fibroblasts. Cell Death Differ. 2002, 9 (2): 125-137. 10.1038/sj.cdd.4400968.
29.
McGowan SE, Torday JS: The pulmonary lipofibroblast (lipid interstitial cell) and its contributions to alveolar development. Annu Rev Physiol. 1997, 59: 43-62. 10.1146/annurev.physiol.59.1.43.
30.
Fries KM, Blieden T, Looney RJ, Sempowski GD, Silvera MR, Willis RA, Phipps RP: Evidence of fibroblast heterogeneity and the role of fibroblast subpopulations in fibrosis. Clin Immunol Immunopathol. 1994, 72 (3): 283-292. 10.1006/clin.1994.1144.
31.
Bradley KH, Kawanami O, Ferrans VJ, Crystal RG: The fibroblast of human lung alveolar structures: a differentiated cell with a major role in lung structure and function. Methods Cell Biol. 1980, 21A: 37-64.
32.
Rose JE, Levin ED, Benowitz N: Saliva nicotine as an index of plasma levels in nicotine skin patch users. Ther Drug Monit. 1993, 15 (5): 431-435. 10.1097/00007691-199310000-00012.
33.
James JA, Sayers NM, Drucker DB, Hull PS: Effects of tobacco products on the attachment and growth of periodontal ligament fibroblasts. J Periodontol. 1999, 70 (5): 518-525. 10.1902/jop.1999.70.5.518.
34.
Vellappally S, Fiala Z, Smejkalova J, Jacob V, Somanathan R: Smoking related systemic and oral diseases. Acta Med (Hradec Kralove). 2007, 50 (3): 161-166.
35.
Lieberman E, Gremy I, Lang JM, Cohen AP: Low birthweight at term and the timing of fetal exposure to maternal smoking. Am J Public Health. 1994, 84 (7): 1127-1131. 10.2105/AJPH.84.7.1127.
36.
Yanagita M, Kojima Y, Kawahara T, Kajikawa T, Oohara H, Takedachi M, Yamada S, Murakami S: Suppressive effects of nicotine on the cytodifferentiation of murine periodontal ligament cells. Oral Dis. 2010, 16 (8): 812-817. 10.1111/j.1601-0825.2010.01693.x.
37.
Somerman MJ, Archer SY, Imm GR, Foster RA: A comparative study of human periodontal ligament cells and gingival fibroblasts in vitro. J Dent Res. 1988, 67 (1): 66-70. 10.1177/00220345880670011301.
38.
Lekic PC, Pender N, McCulloch CA: Is fibroblast heterogeneity relevant to the health, diseases, and treatments of periodontal tissues?. Crit Rev Oral Biol Med. 1997, 8 (3): 253-268. 10.1177/10454411970080030201.
39.
McCulloch CA, Lekic P, McKee MD: Role of physical forces in regulating the form and function of the periodontal ligament. Periodontol 2000. 2000, 24: 56-72. 10.1034/j.1600-0757.2000.2240104.x.
40.
Alpar B, Leyhausen G, Sapotnick A, Gunay H, Geurtsen W: Nicotine-induced alterations in human primary periodontal ligament and gingiva fibroblast cultures. Clin Oral Investig. 1998, 2 (1): 40-46. 10.1007/s007840050042.
41.
Chang HY, Yang X: Proteases for cell suicide: functions and regulation of caspases. Microbiol Mol Biol Rev. 2000, 64 (4): 821-846. 10.1128/MMBR.64.4.821-846.2000.
42.
Kresch MJ, Christian C, Wu F, Hussain N: Ontogeny of apoptosis during lung development. Pediatr Res. 1998, 43 (3): 426-431. 10.1203/00006450-199803000-00020.
43.
Martin TR, Hagimoto N, Nakamura M, Matute-Bello G: Apoptosis and epithelial injury in the lungs. Proc Am Thorac Soc. 2005, 2 (3): 214-220. 10.1513/pats.200504-031AC.
44.
Drakopanagiotakis F, Xifteri A, Polychronopoulos V, Bouros D: Apoptosis in lung injury and fibrosis. Eur Respir J. 2008, 32 (6): 1631-1638. 10.1183/09031936.00176807.
45.
Kumar S: Caspase function in programmed cell death. Cell Death Differ. 2007, 14 (1): 32-43. 10.1038/sj.cdd.4402060.
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