ABSTRACT
Changes in plasma von Willebrand factor concentration (VWF:Ag) and ADAMTS-13 activity (the metalloprotease that cleaves VWF physiologically) have been reported in several cardiovascular disorders with prognostic implications. We therefore determined the level of these proteins in the plasma of children with cyanotic congenital heart disease (CCHD) undergoing surgical treatment. Forty-eight children were enrolled (age 0.83 to 7.58 years). Measurements were performed at baseline and 48 h after surgery. ELISA, collagen-binding assays and Western blotting were used to estimate antigenic and biological activities, and proteolysis of VWF multimers. Preoperatively, VWF:Ag and ADAMTS-13 activity were decreased (65 and 71% of normal levels considered as 113 (105-129) U/dL and 91 ± 24% respectively, P < 0.003) and correlated (r = 0.39, P = 0.0064). High molecular weight VWF multimers were not related, suggesting an interaction of VWF with cell membranes, followed by proteolytic cleavage. A low preoperative ADAMTS-13 activity, a longer activated partial thromboplastin time and the need for cardiopulmonary bypass correlated with postoperative bleeding (P < 0.05). Postoperatively, ADAMTS-13 activity increased but less extensively than VWF:Ag (respectively, 2.23 and 2.83 times baseline, P < 0.0001), resulting in an increased VWF:Ag/ADAMTS-13 activity ratio (1.20 to 1.54, respectively, pre- and postoperative median values, P = 0.0029). ADAMTS-13 consumption was further confirmed by decreased ADAMTS-13 antigenic concentration (0.91 ± 0.30 to 0.70 ± 0.25 µg/mL, P < 0.0001) and persistent proteolysis of VWF multimers. We conclude that, in pediatric CCHD, changes in circulating ADAMTS-13 suggest enzyme consumption, associated with abnormal structure and function of VWF.
Subject(s)
ADAM Proteins/blood , Heart Defects, Congenital/blood , von Willebrand Factor/analysis , ADAMTS13 Protein , Biomarkers/blood , Blotting, Western , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Heart Defects, Congenital/surgery , Humans , Infant , Predictive Value of TestsABSTRACT
Changes in plasma von Willebrand factor concentration (VWF:Ag) and ADAMTS-13 activity (the metalloprotease that cleaves VWF physiologically) have been reported in several cardiovascular disorders with prognostic implications. We therefore determined the level of these proteins in the plasma of children with cyanotic congenital heart disease (CCHD) undergoing surgical treatment. Forty-eight children were enrolled (age 0.83 to 7.58 years). Measurements were performed at baseline and 48 h after surgery. ELISA, collagen-binding assays and Western blotting were used to estimate antigenic and biological activities, and proteolysis of VWF multimers. Preoperatively, VWF:Ag and ADAMTS-13 activity were decreased (65 and 71% of normal levels considered as 113 (105-129) U/dL and 91 ± 24% respectively, P < 0.003) and correlated (r = 0.39, P = 0.0064). High molecular weight VWF multimers were not related, suggesting an interaction of VWF with cell membranes, followed by proteolytic cleavage. A low preoperative ADAMTS-13 activity, a longer activated partial thromboplastin time and the need for cardiopulmonary bypass correlated with postoperative bleeding (P < 0.05). Postoperatively, ADAMTS-13 activity increased but less extensively than VWF:Ag (respectively, 2.23 and 2.83 times baseline, P < 0.0001), resulting in an increased VWF:Ag/ADAMTS-13 activity ratio (1.20 to 1.54, respectively, pre- and postoperative median values, P = 0.0029). ADAMTS-13 consumption was further confirmed by decreased ADAMTS-13 antigenic concentration (0.91 ± 0.30 to 0.70 ± 0.25 µg/mL, P < 0.0001) and persistent proteolysis of VWF multimers. We conclude that, in pediatric CCHD, changes in circulating ADAMTS-13 suggest enzyme consumption, associated with abnormal structure and function of VWF.
Subject(s)
Child , Child, Preschool , Humans , Infant , ADAM Proteins/blood , Heart Defects, Congenital/blood , von Willebrand Factor/analysis , Blotting, Western , Biomarkers/blood , Enzyme-Linked Immunosorbent Assay , Heart Defects, Congenital/surgery , Predictive Value of TestsABSTRACT
The vasorelaxantion of the aqueous fraction of the hydroalcoholic extract of the Sida cordifolia leaves (AFSC) was evaluated in this work. In rat superior mesenteric artery, AFSC (3-1000 microg/mL) induced relaxation of phenylephrine-induced contractions. This effect was significantly attenuated after removal of the endothelium, after atropine (1 microM), L-NAME (100 microM), indomethacin (10 microM), high K+ (20 mM), tetraethylammonium (1 microM), a K(Ca) blocker, apamin (1 microM), a SK(Ca) blocker and ChTX (0.1 microM), a BK(Ca) blocker, however, it was not affected after glibenclamide (10 microM), an KATP blocker, and 4-aminopyridine (1 microM), a Kv blocker. ChTX (0.1 microM) was able to induce an additional inhibition of the vasorelaxation induced by AFSC in the presence of L-NAME plus indomethacin. The vasorelaxation induced by AFSC in the presence of L-NAME plus indomethacin plus ChTX was not different from that induced by AFSC in rings without endothelium. In conclusion, the results show that endothelium-derived factors (mainly NO, PGI2) and K+ channels (BK(Ca) and SK(Ca)) play a crucial role in the vasorelaxation induced by AFSC in the rat superior mesenteric artery.
Subject(s)
Endothelium, Vascular/physiology , Malvaceae/chemistry , Mesenteric Artery, Superior/physiology , Muscle, Smooth, Vascular/physiology , Potassium Channels/physiology , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Charybdotoxin/pharmacology , Endothelium, Vascular/drug effects , Enzyme Inhibitors/pharmacology , In Vitro Techniques , Indomethacin/pharmacology , Mesenteric Artery, Superior/drug effects , Muscle Relaxation/drug effects , Muscle Relaxation/physiology , Muscle, Smooth, Vascular/drug effects , NG-Nitroarginine Methyl Ester/pharmacology , Nitric Oxide Synthase Type III/antagonists & inhibitors , Plant Extracts/pharmacology , Plant Leaves/chemistry , Potassium Channels/drug effects , Potassium Chloride/pharmacology , RatsABSTRACT
The cardiovascular activity of the aqueous fraction of the hydroalcoholic extract of Sida cordifolia leaves (AFSC) was evaluated. In normotensive non-anaesthetized rats was observed that AFSC (5, 10, 20, 30 and 40 mg/kg, i.v.) induced hypotension (6 +/- 2%; 8 +/- 2%; 11 +/- 2%; 19 +/- 3% and 33 +/- 3%, respectively) and bradycardia (0.3 +/- 3%; 13 +/- 4%; 38 +/- 6%; 64 +/- 7% and 80 +/- 5%, respectively). Hypotensive response was completely abolished after atropine (2 mg/kg; i.v.) but potentialized after hexamethonium (20 mg/kg; i.v.) (12 +/- 2%; 21 +/- 5%; 28 +/- 3%; 32 +/- 2% and 32 +/- 3%, respectively), while bradycardic response was completely abolished after atropine (2 mg/kg; i.v.) and attenuated with hexamethonium (20 mg/kg; i.v.) (1 +/- 0.3%; 5 +/- 1%; 7 +/- 1%; 7 +/- 1% and 10 +/- 1%, respectively). In hexamethonium treated rats, L-NAME significantly attenuated the hypotensive response (9 +/- 2%; 14 +/- 1%; 16 +/- 1%; 16 +/- 2% and 22 +/- 3%, respectively). In normotensive anaesthetized and vagotomized rats, hypotensive and bradycardic responses were significantly attenuated (0.5 +/- 0.2%; 1 +/- 0.4%; 3 +/- 0.6%; 4 +/- 0.8% and 6 +/- 1%, respectively, n = 6, and 7 +/- 2%; 12 +/- 5%; 15 +/- 2%, 17 +/- 2% and 25 +/- 3%, respectively). The anaesthesia with sodium thiopental did not affect the AFSC-induced responses when compared with those induced in non-anaesthetized rats (data not showed). In conclusion, the results obtained so far show that AFSC produce hypotension and bradycardia, mainly due to a direct stimulation of the endothelial vascular muscarinic receptor and indirect cardiac muscarinic activation, respectively.
