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Simvastatin reduces monocyte-tissue-factor expression type IIa hypercholesterolaemia
THE LANCET support and advice. This work was supported by a Clinician Investigator Development Award (MK) from the National Institute of Neurological Disorders and Stroke, the Royalty Research Fund of the University of Washington, the Dana Foundation, and National Institutes of Health Grant T32NS-07144-15. 1 2
3
4 5
Filler AG, Howe FA, Hayes CE, et al. Magnetic resonance neurography. Lancet 1993; 341: 659–61. Teresi L, Hovda D, Seeley A, Nitta K, Lufkin R. MR imaging of experimental demyelination. Am J Roentgenol 1989; 152: 1291–98. Titelbaum DS, Frazier JL, Grossman RI, et al. Wallerian degeneration and inflammation in rat peripheral nerve detected by in vivo MR imaging. AJNR Am J Neuroradiol 1989; 10: 741`-46. Does M, Snyder R. Multiexponential T2 relaxation in degenerating peripheral nerve. Magn Reson Med 1996; 35: 207–13. Kline DG, Hudson AR. Nerve injuries: operative results for major nerve injuries, entrapments, and tumors. Philadelphia: WB Saunders Company, 1995.
Department of Neurological Surgery, University of Washington, and VA Puget Sound Health Care System, Box 358 280, 1660 South Columbian Way, Seattle, WA 98108, USA (M Kliot); Department of Radiology, University of Utah, Salt Lake City, Utah; Department of Neurological Surgery, UCLA Medical Center, Los Angeles; and Department of Radiology, University of Washington, Seattle
Simvastatin reduces monocytetissue-factor expression type IIa hypercholesterolaemia Domenico Ferro, Stefania Basili, Cesare Alessandri, Barbara Mantovani, Corrado Cordova, Francesco Violi
Inhibitors of 3-hydroxy-methyl-glutaryl coenzyme A reductase have been shown to reduce cardiovascular events in patients with raised and average serum cholesterol.1,2 It is unclear whether this effect is attributable solely to reduction of serum cholesterol or to other mechanisms.3 We analysed whether simvastatin directly affects expression of monocyte tissue factor (TF). After 4 weeks of diet alone (fat intake <30% of total calories, cholesterol <300 mg daily, polyunsaturated/ saturated fatty acids ratio=1·0) (run-in phase), 24 patients (12 men and 12 women; mean age 52 [SD 8] range 35–70 years) with polygenic hypercholesterolaemia were randomised to simvastatin (20 mg daily) or to continue diet for 8 weeks. Before and after treatment, expression of TF antigen and activity as well as the circulating concentrations of prothrombin fragment F1+2, a marker of thrombin generation, were measured. Nine parts of blood were mixed in a pre-cooled
Assay
vacutainer (Becton Dickinson Vacutainer System, France) with 1 part of 3·8% sodium citrate and centrifuged for 20 min at 2000 g at –4ºC. Plasma samples for measurement of F1+2 (Enzygnost F1+2, Behringwerke ag, Marburg) were stored at –40ºC. To measure monocyte TF, peripheral blood mononuclear cells were isolated from heparinised venous blood (2⫻105 cells/mL)4 and incubated for 6 h with 0·4 ng/mL lipopolysacchariae (Escherichia coli [OB11:B4, Sigma, St Louis, MI, USA). In the lysate of the cells, TF antigen was measured by an ELISA test (Imubind Tissue Factor ELISA kit, American Diagnostica Inc, Greenwich, CT) and TF activity was determined by measuring monocyte procoagulant activity with one stage clotting assay.5 Hypercholesterolaemic patients showed significantly higher concentrations of F1+2 (mean [SD] 2·2 [0·4] vs 0·6 [0·3] nmol/L: p<0·0001), TF antigen (median [range] 63 [41–89] vs 16 [10–39] pg/2⫻105 monocytes; p<0·0001) and activity (median [range] 31 [20–44] vs 9 [4–20] U/2⫻105 monocytes; p<0·0001) than 24 controls matched for age and sex (unpaired t test and Mann-Whitney U test). Patients randomised to diet or diet plus simvastatin had similar clinical (not shown) and laboratory variables (table). In patients continuing the follow-up by diet alone, no significant changes of the variables were observed. In patients taking simvastatin there was a significant decrease of cholesterol, LDL cholesterol, F1+2, and monocyte TF expression. Comparing the diet and simvastatin groups at the end of the treatment period, simvastatin group showed significantly lower concentrations of F1+2, TF antigen, and activity. This study shows that simvastatin reduces the expression of monocyte TF, an effect which may explain the reduction of cardiovascular events elicited by simvastatin. 1
2
3 4
5
Scandinavian Simvastatin Survival Study Group. Randomised trial of cholesterol lowering in 4444 patients with coronary heart disease: the Scandinavian Simvastatin Survival Study (4S). Lancet 1994; 344: 1383–89. Sacks FM, Pfeffer MA, Moye LA, et al. The effect of pravastatin on coronary events after myocardial infarction in patients with average cholesterol levels. N Engl J Med 1996; 335: 1001–09. Vaughan CJ, Murphy MB, Buckley BM. Statins do more than just lower cholesterol. Lancet 1996; 348: 1079–82. Kornberg A, Blank M, Kaufman S, Shoenfeld Y. Induction of tissue factor-like activity in monocytes by anti-cardiolipin antibodies. J Immunol 1994; 153: 1328–32. Semeraro N, Biondi A, Lorenzet R, Locati D, Mantovani A, Donati MB. Direct induction of tissue factor synthesis by endotoxin in human macrophages from diverse anatomical sites. Immunology 1983; 50: 529–35.
Istituto di I Clinica Medica (F Violi), Istituto di Terapia Medica, and Istituto di Terapia Medica Sistematica, Università La Sapienza, 00185 Rome, Italy
Diet plus simvastatin
Diet
p†
Baseline (n=12)
p*
8th week (n=12)
Baseline (n=12)
p*
8th week (n=12)
Total cholesterol (mg/dL)
283·5 (226–331)
<0·003
262 (200–338)
>0·05
Triglycerides (mg/dL)
138·5 (60–305)
<0·02
128·0 (48–207)
>0·05
HDL-cholesterol (mg/dL)
47·5 (28–70)
>0·05
54·0 (37–77)
>0·05
LDL-cholesterol (mg/dL)
206 (154–259)
<0·003
183 (127–261)
>0·05
2·2 (1·8–3·5)
<0·003
2·0 (1·7–2·9)
>0·05
70·6 (50·8–83·6)
<0·003
56·9 (41·0–88·8)
>0·05
32·0 (26–42)
<0·003
208·0 (161–255) ⫺24% 70·0 (44–248) ⫺11% 53·0 +11% 137 (71–179) ⫺28% 1·4 (1·2–1·8) ⫺35% 22·5 (10·4–43·0) ⫺68% 11·9 (3–27) 61%
29 (20–44)
>0·05
268 (200–341) +1% 123·0 (70–206) +8% 55·0 (37–65) +1% 182 (121–270) +2% (1·6–2·6) 2·0 ⫺8% 63·0 (33·9–79·5) ⫺1% 32 (17–42) ⫺4%
F1+2 (nmol/L) Tissue-factor antigen (pg/2⫻105 monocytes) Tissue-factor activity (U/2⫻105 monocytes)
0·002 0·544 0·284 0·006 0·001 0·0002 0·0002
Data are median (range). Values expressed to percentage represent the percentage changes. HDL=high density lipoprotein, LDL=low density lipoprotein, F1+2=prothrombin fragment F1+2. *p-value of Wilcoxon signed rank test. †p-value of Mann-Whitney test used to compare the percentage change from baseline between groups after treatment.
Results in hypercholesterolaemic patients treated for 8 weeks with diet or diet plus simvastatin
1222
Vol 350 • October 25, 1997
3
4 5
Filler AG, Howe FA, Hayes CE, et al. Magnetic resonance neurography. Lancet 1993; 341: 659–61. Teresi L, Hovda D, Seeley A, Nitta K, Lufkin R. MR imaging of experimental demyelination. Am J Roentgenol 1989; 152: 1291–98. Titelbaum DS, Frazier JL, Grossman RI, et al. Wallerian degeneration and inflammation in rat peripheral nerve detected by in vivo MR imaging. AJNR Am J Neuroradiol 1989; 10: 741`-46. Does M, Snyder R. Multiexponential T2 relaxation in degenerating peripheral nerve. Magn Reson Med 1996; 35: 207–13. Kline DG, Hudson AR. Nerve injuries: operative results for major nerve injuries, entrapments, and tumors. Philadelphia: WB Saunders Company, 1995.
Department of Neurological Surgery, University of Washington, and VA Puget Sound Health Care System, Box 358 280, 1660 South Columbian Way, Seattle, WA 98108, USA (M Kliot); Department of Radiology, University of Utah, Salt Lake City, Utah; Department of Neurological Surgery, UCLA Medical Center, Los Angeles; and Department of Radiology, University of Washington, Seattle
Simvastatin reduces monocytetissue-factor expression type IIa hypercholesterolaemia Domenico Ferro, Stefania Basili, Cesare Alessandri, Barbara Mantovani, Corrado Cordova, Francesco Violi
Inhibitors of 3-hydroxy-methyl-glutaryl coenzyme A reductase have been shown to reduce cardiovascular events in patients with raised and average serum cholesterol.1,2 It is unclear whether this effect is attributable solely to reduction of serum cholesterol or to other mechanisms.3 We analysed whether simvastatin directly affects expression of monocyte tissue factor (TF). After 4 weeks of diet alone (fat intake <30% of total calories, cholesterol <300 mg daily, polyunsaturated/ saturated fatty acids ratio=1·0) (run-in phase), 24 patients (12 men and 12 women; mean age 52 [SD 8] range 35–70 years) with polygenic hypercholesterolaemia were randomised to simvastatin (20 mg daily) or to continue diet for 8 weeks. Before and after treatment, expression of TF antigen and activity as well as the circulating concentrations of prothrombin fragment F1+2, a marker of thrombin generation, were measured. Nine parts of blood were mixed in a pre-cooled
Assay
vacutainer (Becton Dickinson Vacutainer System, France) with 1 part of 3·8% sodium citrate and centrifuged for 20 min at 2000 g at –4ºC. Plasma samples for measurement of F1+2 (Enzygnost F1+2, Behringwerke ag, Marburg) were stored at –40ºC. To measure monocyte TF, peripheral blood mononuclear cells were isolated from heparinised venous blood (2⫻105 cells/mL)4 and incubated for 6 h with 0·4 ng/mL lipopolysacchariae (Escherichia coli [OB11:B4, Sigma, St Louis, MI, USA). In the lysate of the cells, TF antigen was measured by an ELISA test (Imubind Tissue Factor ELISA kit, American Diagnostica Inc, Greenwich, CT) and TF activity was determined by measuring monocyte procoagulant activity with one stage clotting assay.5 Hypercholesterolaemic patients showed significantly higher concentrations of F1+2 (mean [SD] 2·2 [0·4] vs 0·6 [0·3] nmol/L: p<0·0001), TF antigen (median [range] 63 [41–89] vs 16 [10–39] pg/2⫻105 monocytes; p<0·0001) and activity (median [range] 31 [20–44] vs 9 [4–20] U/2⫻105 monocytes; p<0·0001) than 24 controls matched for age and sex (unpaired t test and Mann-Whitney U test). Patients randomised to diet or diet plus simvastatin had similar clinical (not shown) and laboratory variables (table). In patients continuing the follow-up by diet alone, no significant changes of the variables were observed. In patients taking simvastatin there was a significant decrease of cholesterol, LDL cholesterol, F1+2, and monocyte TF expression. Comparing the diet and simvastatin groups at the end of the treatment period, simvastatin group showed significantly lower concentrations of F1+2, TF antigen, and activity. This study shows that simvastatin reduces the expression of monocyte TF, an effect which may explain the reduction of cardiovascular events elicited by simvastatin. 1
2
3 4
5
Scandinavian Simvastatin Survival Study Group. Randomised trial of cholesterol lowering in 4444 patients with coronary heart disease: the Scandinavian Simvastatin Survival Study (4S). Lancet 1994; 344: 1383–89. Sacks FM, Pfeffer MA, Moye LA, et al. The effect of pravastatin on coronary events after myocardial infarction in patients with average cholesterol levels. N Engl J Med 1996; 335: 1001–09. Vaughan CJ, Murphy MB, Buckley BM. Statins do more than just lower cholesterol. Lancet 1996; 348: 1079–82. Kornberg A, Blank M, Kaufman S, Shoenfeld Y. Induction of tissue factor-like activity in monocytes by anti-cardiolipin antibodies. J Immunol 1994; 153: 1328–32. Semeraro N, Biondi A, Lorenzet R, Locati D, Mantovani A, Donati MB. Direct induction of tissue factor synthesis by endotoxin in human macrophages from diverse anatomical sites. Immunology 1983; 50: 529–35.
Istituto di I Clinica Medica (F Violi), Istituto di Terapia Medica, and Istituto di Terapia Medica Sistematica, Università La Sapienza, 00185 Rome, Italy
Diet plus simvastatin
Diet
p†
Baseline (n=12)
p*
8th week (n=12)
Baseline (n=12)
p*
8th week (n=12)
Total cholesterol (mg/dL)
283·5 (226–331)
<0·003
262 (200–338)
>0·05
Triglycerides (mg/dL)
138·5 (60–305)
<0·02
128·0 (48–207)
>0·05
HDL-cholesterol (mg/dL)
47·5 (28–70)
>0·05
54·0 (37–77)
>0·05
LDL-cholesterol (mg/dL)
206 (154–259)
<0·003
183 (127–261)
>0·05
2·2 (1·8–3·5)
<0·003
2·0 (1·7–2·9)
>0·05
70·6 (50·8–83·6)
<0·003
56·9 (41·0–88·8)
>0·05
32·0 (26–42)
<0·003
208·0 (161–255) ⫺24% 70·0 (44–248) ⫺11% 53·0 +11% 137 (71–179) ⫺28% 1·4 (1·2–1·8) ⫺35% 22·5 (10·4–43·0) ⫺68% 11·9 (3–27) 61%
29 (20–44)
>0·05
268 (200–341) +1% 123·0 (70–206) +8% 55·0 (37–65) +1% 182 (121–270) +2% (1·6–2·6) 2·0 ⫺8% 63·0 (33·9–79·5) ⫺1% 32 (17–42) ⫺4%
F1+2 (nmol/L) Tissue-factor antigen (pg/2⫻105 monocytes) Tissue-factor activity (U/2⫻105 monocytes)
0·002 0·544 0·284 0·006 0·001 0·0002 0·0002
Data are median (range). Values expressed to percentage represent the percentage changes. HDL=high density lipoprotein, LDL=low density lipoprotein, F1+2=prothrombin fragment F1+2. *p-value of Wilcoxon signed rank test. †p-value of Mann-Whitney test used to compare the percentage change from baseline between groups after treatment.
Results in hypercholesterolaemic patients treated for 8 weeks with diet or diet plus simvastatin
1222
Vol 350 • October 25, 1997