1979,Vol. 37, pp. 367369, O Hippokrates Verlag GmbH
Journald
medica
~ b ne~esearth t d i
Inhibition of Platelet Aggregation by Guggulu" Steroids
I I
L. Mester*, M. Mester* and Swarn Nityanand** lnstitut de Chimie des Substances Naturelles, Gif-sur-Yvette, France.
Key Word Index: Commiphora mukul; Burseraceae; Platelet Aggregation; ADP; Adrenaline; S,erotonin; Clofibrate; Steroids.
obesity and allied disorders [2]. Recent pharmacological studies have revealed Purified steroidal fractions of ,guggu- in addition hypocholesterolemic and lua show a marked inhibition of ADP, hypolipaemic activity [3-71. adrenaline and serotonin induced plateAmongst many compounds isolated let aggregation. Their inhibitory effect from this resin are two steroids:guggulis comparable to the .effect o f ,Clofi- sterones E (I) and Z (11), cis and trans isomers of 4 , 17 (20)-pregna-3, 16&ratea. dione (8). The hypolipaemic activity of these Introduction two steroids and lipolysis in fat cells studies by adrenaline induced FFA reThe ayurvedic crude drug ,gugguluU lease in the presence of lipid lowering is the gum-resin exudate from the plant agent both in vitro and in vivo and lipoCornmiphora mukul. According to genesis as assessed by in vitro inhibition Ayurveda [I], ,guggulua i,s effective in of cholesterol biosynthesis using acetate in liver homogenates, was signithe treatment of rheumatoid arthritis, I-c14
Abstract
Downloaded by: University of Pennsylvania Libraries. Copyrighted material.
" Central Drug Research Institute, Lucknow, India.
ficant and comparable to clofibrate as reported earlier [9-111. Recently, it has been reported that testosteron increases, while estradiol, progesterone and desoxycorticosterone decreases ADP induced platelet aggregation [I21. Therefore, it was considered of interest to study the effect of guggul-sterones E and Z in platelet aggregation. Material and Methods Crude fractions of ,gugguluU and pure E and Z guggul-sterones were prepared as reported [8]. Platelet-rich plasma (PRP) was obtained
from blood collected on hypercitrated anticoagulant [I31 and adjusted to a concentration of 3 X lo8 platelets/ml. Platelet aggregation induced by ADP . a t a final conc. of M, by adrenaline 1.2X M and by serotonin oxalate lo-' M, was measured using a ,Labintecu (Montpellier, France) aggregometer. To 0.3 ml citrated PRP a pH = 7.4 MiAaeIis buffer solution was added to make up a final volume of 0.4 mI, after addition of the test solution. For investination of the inhibitory effect, the compounds were added to the PRPfbuffer mixture 5 min. before the addition of the aggregating agent. The PRPf bufferfinhibitor mixture was first heated for 3 min. at 37O C, than stirred for 2 min. The inhibitory effect was expressed as 010 decrease of the total aggregation observed.
Table I. Platelet Aggregation induced by 'Inhibitor
Final conc.
ADP l x 1OmSM final conc.
Adrenaline 1 . 2 1~0 ' ~ final conc.
Serotonin 1X I O - ~ M final conc .
Control Adenosine Guggulu EtOAc Fr. Steroid Mixtr.
Clofibrate
Ix
IO‘~M
30
20
20
Downloaded by: University of Pennsylvania Libraries. Copyrighted material.
Mester, L., M. Mester, Nityanand
Results and Discussion
I t is reported for the first time a marked inhibition of ADP, adrenaline and serotonin induced platelet aggregation with purified steroidal fractions of ,gugguluU. The purified steroid mixture a t 2X I O - ~final M concentration inhibited completely ADP, adrenaline or serotonin induced platelet aggregation. N o difference has been observed between the effectiveness of the steroid mixture and the purified guggulsterone-E or guggulsterone-2, respectively. The effect of isolated guggulsterone E and 2 a t I O - final ~ M concentration on ADP, adrenaline or serotonin induced platelet aggregation is very similar to the inhibitory effect of ,clofibratea at the same final concentration. This finding has its therapeutic value in myocardial infarction and thromboernbolism.
Acknowledgement A part of this work was realized in the framework of the Indo-French Scicntific Cooperation Prograrnme.The authors are indebted to Dr. SUKHDEVfor supplying steroid samples and to Dr. NITYAANANDfor useful advises.
References I. A W R V E D A is the ancient Indian system of treating body disorders.
369
2. Vd. Bhagwan Dash, "Ayurvedic Treatment for Common Diseases" p. 112 and 127. New Delhi, 1974, Delhi Diary Publ. 3. Mehta, V. L., C. L. Malhotra and S. N. Kalrah: Indian J. Physiol. and Pharmacol., 12, 91 (1968). 4. Sathavati, G. V., C. Dwarakanath and S. N. Tripathi: Indian J. Med. Res., 57, 1950 (1969). 5. Malhotra, C. L., Y.K. Agarwal, V. L. Mehta and S. Prasad: Indian J. Med. Res., J8, 394 (1970). 6. Kapoor, N. K. and S. Nityanand: .Hypocholestremic Effect of the Fraction isolated from Commophora mukul (Guggulu)" Seminar on Disorders of Lipid Metabolism, New Delhi, 1971. 7. Nityanand, S. and N. K. Kapoor: .Cholesterol lowering activity of the various Fractions of Commiphora mukul (Guggulu)", Indian J. Exp. Biol., 11, 395 (1973). 8. Patil, V. D., U. R. Nayak and Dev Sukh: Tetrahedron, 28, 2341 (1972). 9. Kapoor, N. K., S.Nityanand and Dev Sukh: ,Commiphora mukul and its different extracts as hypolipidaemic agents" Indo-Soviet Symposium on Chemistry of Natural Products, Central Drug Research Institute, Lucknow, 1976. 10. Kapoor, N. K. and S. Nityanand: "Effect of guggul-steroids on Cholesterol biosynthesis in rats" Ind. J. Bioehem. Biophys., 15, 77 (1978). 11. Data on .GuggulipidU Dossier, C.D.R.I., Lucknow, 1976. 12. Johnson, M., E. Ramey and P. W. Ramwell: Am. J. Phys. 232 (1977). 13. Born, G. V. R. and M. Hume: Nature (London), 215, 1027 (1967).
Adresse: Prof. L. Mester, Centre National de la Reserche Scientifique, Institut de Chimie des Substances Naturelles, 91 190 Gif-Sur-YvettelFrance
Downloaded by: University of Pennsylvania Libraries. Copyrighted material.
Inhibition of Platelet Aggregation by "Guggulu" Steroids
Journald
medica
~ b ne~esearth t d i
Inhibition of Platelet Aggregation by Guggulu" Steroids
I I
L. Mester*, M. Mester* and Swarn Nityanand** lnstitut de Chimie des Substances Naturelles, Gif-sur-Yvette, France.
Key Word Index: Commiphora mukul; Burseraceae; Platelet Aggregation; ADP; Adrenaline; S,erotonin; Clofibrate; Steroids.
obesity and allied disorders [2]. Recent pharmacological studies have revealed Purified steroidal fractions of ,guggu- in addition hypocholesterolemic and lua show a marked inhibition of ADP, hypolipaemic activity [3-71. adrenaline and serotonin induced plateAmongst many compounds isolated let aggregation. Their inhibitory effect from this resin are two steroids:guggulis comparable to the .effect o f ,Clofi- sterones E (I) and Z (11), cis and trans isomers of 4 , 17 (20)-pregna-3, 16&ratea. dione (8). The hypolipaemic activity of these Introduction two steroids and lipolysis in fat cells studies by adrenaline induced FFA reThe ayurvedic crude drug ,gugguluU lease in the presence of lipid lowering is the gum-resin exudate from the plant agent both in vitro and in vivo and lipoCornmiphora mukul. According to genesis as assessed by in vitro inhibition Ayurveda [I], ,guggulua i,s effective in of cholesterol biosynthesis using acetate in liver homogenates, was signithe treatment of rheumatoid arthritis, I-c14
Abstract
Downloaded by: University of Pennsylvania Libraries. Copyrighted material.
" Central Drug Research Institute, Lucknow, India.
ficant and comparable to clofibrate as reported earlier [9-111. Recently, it has been reported that testosteron increases, while estradiol, progesterone and desoxycorticosterone decreases ADP induced platelet aggregation [I21. Therefore, it was considered of interest to study the effect of guggul-sterones E and Z in platelet aggregation. Material and Methods Crude fractions of ,gugguluU and pure E and Z guggul-sterones were prepared as reported [8]. Platelet-rich plasma (PRP) was obtained
from blood collected on hypercitrated anticoagulant [I31 and adjusted to a concentration of 3 X lo8 platelets/ml. Platelet aggregation induced by ADP . a t a final conc. of M, by adrenaline 1.2X M and by serotonin oxalate lo-' M, was measured using a ,Labintecu (Montpellier, France) aggregometer. To 0.3 ml citrated PRP a pH = 7.4 MiAaeIis buffer solution was added to make up a final volume of 0.4 mI, after addition of the test solution. For investination of the inhibitory effect, the compounds were added to the PRPfbuffer mixture 5 min. before the addition of the aggregating agent. The PRPf bufferfinhibitor mixture was first heated for 3 min. at 37O C, than stirred for 2 min. The inhibitory effect was expressed as 010 decrease of the total aggregation observed.
Table I. Platelet Aggregation induced by 'Inhibitor
Final conc.
ADP l x 1OmSM final conc.
Adrenaline 1 . 2 1~0 ' ~ final conc.
Serotonin 1X I O - ~ M final conc .
Control Adenosine Guggulu EtOAc Fr. Steroid Mixtr.
Clofibrate
Ix
IO‘~M
30
20
20
Downloaded by: University of Pennsylvania Libraries. Copyrighted material.
Mester, L., M. Mester, Nityanand
Results and Discussion
I t is reported for the first time a marked inhibition of ADP, adrenaline and serotonin induced platelet aggregation with purified steroidal fractions of ,gugguluU. The purified steroid mixture a t 2X I O - ~final M concentration inhibited completely ADP, adrenaline or serotonin induced platelet aggregation. N o difference has been observed between the effectiveness of the steroid mixture and the purified guggulsterone-E or guggulsterone-2, respectively. The effect of isolated guggulsterone E and 2 a t I O - final ~ M concentration on ADP, adrenaline or serotonin induced platelet aggregation is very similar to the inhibitory effect of ,clofibratea at the same final concentration. This finding has its therapeutic value in myocardial infarction and thromboernbolism.
Acknowledgement A part of this work was realized in the framework of the Indo-French Scicntific Cooperation Prograrnme.The authors are indebted to Dr. SUKHDEVfor supplying steroid samples and to Dr. NITYAANANDfor useful advises.
References I. A W R V E D A is the ancient Indian system of treating body disorders.
369
2. Vd. Bhagwan Dash, "Ayurvedic Treatment for Common Diseases" p. 112 and 127. New Delhi, 1974, Delhi Diary Publ. 3. Mehta, V. L., C. L. Malhotra and S. N. Kalrah: Indian J. Physiol. and Pharmacol., 12, 91 (1968). 4. Sathavati, G. V., C. Dwarakanath and S. N. Tripathi: Indian J. Med. Res., 57, 1950 (1969). 5. Malhotra, C. L., Y.K. Agarwal, V. L. Mehta and S. Prasad: Indian J. Med. Res., J8, 394 (1970). 6. Kapoor, N. K. and S. Nityanand: .Hypocholestremic Effect of the Fraction isolated from Commophora mukul (Guggulu)" Seminar on Disorders of Lipid Metabolism, New Delhi, 1971. 7. Nityanand, S. and N. K. Kapoor: .Cholesterol lowering activity of the various Fractions of Commiphora mukul (Guggulu)", Indian J. Exp. Biol., 11, 395 (1973). 8. Patil, V. D., U. R. Nayak and Dev Sukh: Tetrahedron, 28, 2341 (1972). 9. Kapoor, N. K., S.Nityanand and Dev Sukh: ,Commiphora mukul and its different extracts as hypolipidaemic agents" Indo-Soviet Symposium on Chemistry of Natural Products, Central Drug Research Institute, Lucknow, 1976. 10. Kapoor, N. K. and S. Nityanand: "Effect of guggul-steroids on Cholesterol biosynthesis in rats" Ind. J. Bioehem. Biophys., 15, 77 (1978). 11. Data on .GuggulipidU Dossier, C.D.R.I., Lucknow, 1976. 12. Johnson, M., E. Ramey and P. W. Ramwell: Am. J. Phys. 232 (1977). 13. Born, G. V. R. and M. Hume: Nature (London), 215, 1027 (1967).
Adresse: Prof. L. Mester, Centre National de la Reserche Scientifique, Institut de Chimie des Substances Naturelles, 91 190 Gif-Sur-YvettelFrance
Downloaded by: University of Pennsylvania Libraries. Copyrighted material.
Inhibition of Platelet Aggregation by "Guggulu" Steroids
