Doxycycline inhibits elastin degradation and reduces metalloproteinase activity in a model of aneurysmal disease Jonathan R. Boyle, FRCS, Edward McDermott,

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Presentation transcript:

Doxycycline inhibits elastin degradation and reduces metalloproteinase activity in a model of aneurysmal disease Jonathan R. Boyle, FRCS, Edward McDermott, BSc, Matthew Crowther, MSc, Andrew D. Wills, BSc, Peter R.F. Bell, MD, FRCS, Matthew M. Thompson, MD, FRCS Journal of Vascular Surgery Volume 27, Issue 2, Pages 354-361 (February 1998) DOI: 10.1016/S0741-5214(98)70367-2 Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Four histologic sections from the same aorta stained with Miller's elastin and van Gieson's stain (original magnification, 400×). Cultured control (A) and three sections exposed to a pulse of elastase, one subsequently cultured in standard medium (B), one in standard medium supplemented by doxycycline 1 mg/L (C), and one in standard medium supplemented by doxycycline 10 mg/L .. Adventitial collagen (D) is noted to be particularly well preserved, although the significance of this is unknown. Journal of Vascular Surgery 1998 27, 354-361DOI: (10.1016/S0741-5214(98)70367-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Graph plots percentage elastin concentration (median values and Q3) for fresh aorta (Fresh), cultured control (CC), elastase exposed after 24 hours (E1), elastase exposed after 14 days (E14), doxycycline-treated 1 mg/L (D1), and doxycycline-treated 10 mg/L (D10). Journal of Vascular Surgery 1998 27, 354-361DOI: (10.1016/S0741-5214(98)70367-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Gelatin zymogram shows metalloproteinase activity for all sections of one aorta and demonstrates reduction in MMP-9 activity in doxycycline-treated segments. Positive C, HT 1080 fibrosarcoma cell line, which produces large quantities of MMP-2 and MMP-9. Fresh, Freshly harvested, noncultured aortic sample. Cultured C, Aorta cultured without exposure to elastase for 14 days. Elastase, Aorta exposed to elastase for 24 hours and then cultured in standard conditions for a further 13 days. Doxy 1 mg/l and Doxy 10 mg/l, Aortas exposed to elastase for 24 hours and then cultured with doxycycline for 13 days. Journal of Vascular Surgery 1998 27, 354-361DOI: (10.1016/S0741-5214(98)70367-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Graph plots MMP-9 activity (median values and Q3) as a fraction of cultured control (CC), confirming that elastase exposure induced MMP-9 activity (E) that was progressively inhibited by an increasing dose of doxycycline at 1 mg/L and 10 mg/L (D1 and D10). Journal of Vascular Surgery 1998 27, 354-361DOI: (10.1016/S0741-5214(98)70367-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Immunoblot demonstrates TIMP-1 immunoreactivity. Cultured C, Aorta cultured without exposure to elastase for 14 days. Elastase, Aorta exposed to elastase for 24 hours and then cultured in standard conditions for a further 13 days. Doxy 1 mg/l and Doxy 10 mg/l, Aortas exposed to elastase for 24 hours and then cultured with doxycycline for 13 days. Positive C, HT 1080 fibrosarcoma cell line, which produces large quantities of MMP-2 and MMP-9. Journal of Vascular Surgery 1998 27, 354-361DOI: (10.1016/S0741-5214(98)70367-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions