Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells

Kevin Harkin, Adam Metcalfe, Mark Harbinson, Malcolm Campbell, Paul Spiers, David Bell

Research output: Contribution to conferenceAbstract

Abstract

Intermedin/adrenomedullin-2 (IMD/AM2) is a peptide closely related to adrenomedullin (AM) and calcitonin gene-related peptide, with which it shares a family of receptors (1). IMD/AM2 protects the mammalian heart and vasculature from oxidative stress and ischaemia-reperfusion injury (2). Less is known about the distribution of IMD/AM2 and its receptor components and potential for a similar protective role in the mammalian kidney, particularly in human (1, 3). The aim was to study expression of the peptide and its receptor components in human renal mesangial pericytes (HRMC) and in tubular epithelial cells (HREpiC) and determine protective effects of IMD/AM2 on cells subjected to nephrotoxic drugs. Expression of IMD and receptor components was determined at mRNA level by quantitative RT-PCR, expressed relative to AM, and at protein level by immunofluorescence staining of cells attached to coverslips, expressed relative to actin. Nephrotoxic drugs were applied to confluent cells for 48 h +/- IMD (1 nM). Cellular injury was determined qualitatively by microscopy and quantitatively by trypan blue exclusion. Data are given as mean+SEM (n cell sources) and analysis was performed using two factor ANOVA followed by unpaired Student’s T-test, as applicable. IMD/AM2 and the receptor components (CLR, RAMP1-3) were detected at mRNA level in HREpiC and HRMC (n=6). Expression was confirmed at protein level by immunofluorescence (n=10). Nephrotoxic drugs induced injury (p<0.05) in HRMC and HREpiC at 48 h as evidenced by microscopy. IMD/AM2 did not influence cell viability in the absence of nephrotoxic drugs but did attenuate nephrotoxic drug injury (Table 1). Table 1 Protection by IMD/AM2 (1 nM) from nephrotoxic drug injury; viabilities at 48 h are expressed relative to control in the absence of IMD/AM and nephrotoxic drugs and are mean+SEM, n=5 sources each run in quadruplicate. *p<0.05 nephrotoxic drug vs. untreated cells; #p<0.05 with vs. without IMD. Untreated Lithium 1.5 mM Indomethacin 7 mg.L-1 Epirubicin 2µg.ml-1 HRMC control 100.0+3.97 54.78+3.38* 39.29+2.35* 47.69+5.27* HRMC + IMD (1 nM) 98.44+3.36 79.74+2.24# 61.13+1.06# 60.60+6.56 HREpiC control 100.0+6.34 66.50+5.01* 45.03+2.52* 51.82+4.83* HREpiC + IMD (1 nM) 102.7+3.44 83.48+2.75# 72.61+4.30# 68.52+7.03 In summary, IMD/AM2 is present in human renal cells and is protective of both mesangial pericytes and tubular epithelial cells against injury induced by a range of nephrotoxic drugs. (1) Holmes D et al. (2013). Curr Prot Pept Sci 14: 294-329. (2) Bell D et al. (2012). J Physiol 590: 1181-1197. (3) Qiao X et al. (2013). Am J Physiol-Renal Physiol 304: F112-F119.
Original languageEnglish
PagesPB128
Number of pages1
Publication statusPublished - 13 Dec 2016
EventBritish Pharmacological Society Winter Meeting 2016 - Queen Elizabeth Conference Centre, London, United Kingdom
Duration: 13 Dec 201615 Dec 2016

Conference

ConferenceBritish Pharmacological Society Winter Meeting 2016
CountryUnited Kingdom
CityLondon
Period13/12/201615/12/2016

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Adrenomedullin
Epithelial Cells
Kidney
Pericytes
Wounds and Injuries
Pharmaceutical Preparations
Fluorescent Antibody Technique
Microscopy
Adrenomedullin Receptors
Messenger RNA
Peptide Receptors
Trypan Blue
Calcitonin Gene-Related Peptide
Reperfusion Injury
Actins
Cell Survival
Analysis of Variance
Proteins
Oxidative Stress
Staining and Labeling

Cite this

Harkin, K., Metcalfe, A., Harbinson, M., Campbell, M., Spiers, P., & Bell, D. (2016). Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells. PB128. Abstract from British Pharmacological Society Winter Meeting 2016, London, United Kingdom.
Harkin, Kevin ; Metcalfe, Adam ; Harbinson, Mark ; Campbell, Malcolm ; Spiers, Paul ; Bell, David. / Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells. Abstract from British Pharmacological Society Winter Meeting 2016, London, United Kingdom.1 p.
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title = "Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells",
abstract = "Intermedin/adrenomedullin-2 (IMD/AM2) is a peptide closely related to adrenomedullin (AM) and calcitonin gene-related peptide, with which it shares a family of receptors (1). IMD/AM2 protects the mammalian heart and vasculature from oxidative stress and ischaemia-reperfusion injury (2). Less is known about the distribution of IMD/AM2 and its receptor components and potential for a similar protective role in the mammalian kidney, particularly in human (1, 3). The aim was to study expression of the peptide and its receptor components in human renal mesangial pericytes (HRMC) and in tubular epithelial cells (HREpiC) and determine protective effects of IMD/AM2 on cells subjected to nephrotoxic drugs. Expression of IMD and receptor components was determined at mRNA level by quantitative RT-PCR, expressed relative to AM, and at protein level by immunofluorescence staining of cells attached to coverslips, expressed relative to actin. Nephrotoxic drugs were applied to confluent cells for 48 h +/- IMD (1 nM). Cellular injury was determined qualitatively by microscopy and quantitatively by trypan blue exclusion. Data are given as mean+SEM (n cell sources) and analysis was performed using two factor ANOVA followed by unpaired Student’s T-test, as applicable. IMD/AM2 and the receptor components (CLR, RAMP1-3) were detected at mRNA level in HREpiC and HRMC (n=6). Expression was confirmed at protein level by immunofluorescence (n=10). Nephrotoxic drugs induced injury (p<0.05) in HRMC and HREpiC at 48 h as evidenced by microscopy. IMD/AM2 did not influence cell viability in the absence of nephrotoxic drugs but did attenuate nephrotoxic drug injury (Table 1). Table 1 Protection by IMD/AM2 (1 nM) from nephrotoxic drug injury; viabilities at 48 h are expressed relative to control in the absence of IMD/AM and nephrotoxic drugs and are mean+SEM, n=5 sources each run in quadruplicate. *p<0.05 nephrotoxic drug vs. untreated cells; #p<0.05 with vs. without IMD. Untreated Lithium 1.5 mM Indomethacin 7 mg.L-1 Epirubicin 2µg.ml-1 HRMC control 100.0+3.97 54.78+3.38* 39.29+2.35* 47.69+5.27* HRMC + IMD (1 nM) 98.44+3.36 79.74+2.24# 61.13+1.06# 60.60+6.56 HREpiC control 100.0+6.34 66.50+5.01* 45.03+2.52* 51.82+4.83* HREpiC + IMD (1 nM) 102.7+3.44 83.48+2.75# 72.61+4.30# 68.52+7.03 In summary, IMD/AM2 is present in human renal cells and is protective of both mesangial pericytes and tubular epithelial cells against injury induced by a range of nephrotoxic drugs. (1) Holmes D et al. (2013). Curr Prot Pept Sci 14: 294-329. (2) Bell D et al. (2012). J Physiol 590: 1181-1197. (3) Qiao X et al. (2013). Am J Physiol-Renal Physiol 304: F112-F119.",
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Harkin, K, Metcalfe, A, Harbinson, M, Campbell, M, Spiers, P & Bell, D 2016, 'Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells', British Pharmacological Society Winter Meeting 2016, London, United Kingdom, 13/12/2016 - 15/12/2016 pp. PB128.

Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells. / Harkin, Kevin; Metcalfe, Adam; Harbinson, Mark; Campbell, Malcolm; Spiers, Paul; Bell, David.

2016. PB128 Abstract from British Pharmacological Society Winter Meeting 2016, London, United Kingdom.

Research output: Contribution to conferenceAbstract

TY - CONF

T1 - Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells

AU - Harkin, Kevin

AU - Metcalfe, Adam

AU - Harbinson, Mark

AU - Campbell, Malcolm

AU - Spiers, Paul

AU - Bell, David

PY - 2016/12/13

Y1 - 2016/12/13

N2 - Intermedin/adrenomedullin-2 (IMD/AM2) is a peptide closely related to adrenomedullin (AM) and calcitonin gene-related peptide, with which it shares a family of receptors (1). IMD/AM2 protects the mammalian heart and vasculature from oxidative stress and ischaemia-reperfusion injury (2). Less is known about the distribution of IMD/AM2 and its receptor components and potential for a similar protective role in the mammalian kidney, particularly in human (1, 3). The aim was to study expression of the peptide and its receptor components in human renal mesangial pericytes (HRMC) and in tubular epithelial cells (HREpiC) and determine protective effects of IMD/AM2 on cells subjected to nephrotoxic drugs. Expression of IMD and receptor components was determined at mRNA level by quantitative RT-PCR, expressed relative to AM, and at protein level by immunofluorescence staining of cells attached to coverslips, expressed relative to actin. Nephrotoxic drugs were applied to confluent cells for 48 h +/- IMD (1 nM). Cellular injury was determined qualitatively by microscopy and quantitatively by trypan blue exclusion. Data are given as mean+SEM (n cell sources) and analysis was performed using two factor ANOVA followed by unpaired Student’s T-test, as applicable. IMD/AM2 and the receptor components (CLR, RAMP1-3) were detected at mRNA level in HREpiC and HRMC (n=6). Expression was confirmed at protein level by immunofluorescence (n=10). Nephrotoxic drugs induced injury (p<0.05) in HRMC and HREpiC at 48 h as evidenced by microscopy. IMD/AM2 did not influence cell viability in the absence of nephrotoxic drugs but did attenuate nephrotoxic drug injury (Table 1). Table 1 Protection by IMD/AM2 (1 nM) from nephrotoxic drug injury; viabilities at 48 h are expressed relative to control in the absence of IMD/AM and nephrotoxic drugs and are mean+SEM, n=5 sources each run in quadruplicate. *p<0.05 nephrotoxic drug vs. untreated cells; #p<0.05 with vs. without IMD. Untreated Lithium 1.5 mM Indomethacin 7 mg.L-1 Epirubicin 2µg.ml-1 HRMC control 100.0+3.97 54.78+3.38* 39.29+2.35* 47.69+5.27* HRMC + IMD (1 nM) 98.44+3.36 79.74+2.24# 61.13+1.06# 60.60+6.56 HREpiC control 100.0+6.34 66.50+5.01* 45.03+2.52* 51.82+4.83* HREpiC + IMD (1 nM) 102.7+3.44 83.48+2.75# 72.61+4.30# 68.52+7.03 In summary, IMD/AM2 is present in human renal cells and is protective of both mesangial pericytes and tubular epithelial cells against injury induced by a range of nephrotoxic drugs. (1) Holmes D et al. (2013). Curr Prot Pept Sci 14: 294-329. (2) Bell D et al. (2012). J Physiol 590: 1181-1197. (3) Qiao X et al. (2013). Am J Physiol-Renal Physiol 304: F112-F119.

AB - Intermedin/adrenomedullin-2 (IMD/AM2) is a peptide closely related to adrenomedullin (AM) and calcitonin gene-related peptide, with which it shares a family of receptors (1). IMD/AM2 protects the mammalian heart and vasculature from oxidative stress and ischaemia-reperfusion injury (2). Less is known about the distribution of IMD/AM2 and its receptor components and potential for a similar protective role in the mammalian kidney, particularly in human (1, 3). The aim was to study expression of the peptide and its receptor components in human renal mesangial pericytes (HRMC) and in tubular epithelial cells (HREpiC) and determine protective effects of IMD/AM2 on cells subjected to nephrotoxic drugs. Expression of IMD and receptor components was determined at mRNA level by quantitative RT-PCR, expressed relative to AM, and at protein level by immunofluorescence staining of cells attached to coverslips, expressed relative to actin. Nephrotoxic drugs were applied to confluent cells for 48 h +/- IMD (1 nM). Cellular injury was determined qualitatively by microscopy and quantitatively by trypan blue exclusion. Data are given as mean+SEM (n cell sources) and analysis was performed using two factor ANOVA followed by unpaired Student’s T-test, as applicable. IMD/AM2 and the receptor components (CLR, RAMP1-3) were detected at mRNA level in HREpiC and HRMC (n=6). Expression was confirmed at protein level by immunofluorescence (n=10). Nephrotoxic drugs induced injury (p<0.05) in HRMC and HREpiC at 48 h as evidenced by microscopy. IMD/AM2 did not influence cell viability in the absence of nephrotoxic drugs but did attenuate nephrotoxic drug injury (Table 1). Table 1 Protection by IMD/AM2 (1 nM) from nephrotoxic drug injury; viabilities at 48 h are expressed relative to control in the absence of IMD/AM and nephrotoxic drugs and are mean+SEM, n=5 sources each run in quadruplicate. *p<0.05 nephrotoxic drug vs. untreated cells; #p<0.05 with vs. without IMD. Untreated Lithium 1.5 mM Indomethacin 7 mg.L-1 Epirubicin 2µg.ml-1 HRMC control 100.0+3.97 54.78+3.38* 39.29+2.35* 47.69+5.27* HRMC + IMD (1 nM) 98.44+3.36 79.74+2.24# 61.13+1.06# 60.60+6.56 HREpiC control 100.0+6.34 66.50+5.01* 45.03+2.52* 51.82+4.83* HREpiC + IMD (1 nM) 102.7+3.44 83.48+2.75# 72.61+4.30# 68.52+7.03 In summary, IMD/AM2 is present in human renal cells and is protective of both mesangial pericytes and tubular epithelial cells against injury induced by a range of nephrotoxic drugs. (1) Holmes D et al. (2013). Curr Prot Pept Sci 14: 294-329. (2) Bell D et al. (2012). J Physiol 590: 1181-1197. (3) Qiao X et al. (2013). Am J Physiol-Renal Physiol 304: F112-F119.

M3 - Abstract

SP - PB128

ER -

Harkin K, Metcalfe A, Harbinson M, Campbell M, Spiers P, Bell D. Influence of intermedin/adrenomedullin-2 on nephrotoxic drug-related injury to human renal mesangial and tubular epithelial cells. 2016. Abstract from British Pharmacological Society Winter Meeting 2016, London, United Kingdom.