Pharmacological and Clinical Study on Cystone

Transcription

1 [Probe (1977): (XVII), 1, 25-29] Pharmacological and Clinical Study on Cystone Phukan, D.P., M.D., Professor and Head of the Department of Pharmacology, (Mrs.) Deka, A., M.D., Demonstrator of Pharmacology and Choudhury, A.K., Demonstrator of Pharmacology, Gauhati Medical College, Gauhati, India. [Abstracted from: Journal of the Gauhati Medical College ( ): 9 th Issue, Scientific Section, p. 12.] INTRODUCTION Cystone, an indigenous preparation of The Himalaya Drug Company, Bombay, consisting of Didymocarpus pedicellata, Saxifraga ligulata, Rubia cordifolia, Cyperus scariosus, Achyranthes aspera, Onosma bracteatum, Vernonia cinerea, Shilajeet (purified) and Hajrul Yahood Bhasma has already established a reputation for its efficacy in relieving crystalluria, and dissolving small stones of kidney, urinary bladder and in the urinary tract. Besides, it has also been observed in clinical cases that Cystone possesses a diuretic and antispasmodic effect on the smooth muscles of the ureter and urinary bladder. The mechanism of the action of Cystone is as yet unknown. However, it was decided in this project to carry out the pharmacological study of Cystone for the following effects: 1. Its diuretic activity 2. Changes in electrolytes 3. Effect on crystalloid-colloid balance 4. Relaxation of smooth muscles 5. Other miscellaneous effects MATERIALS AND METHODS In all these investigations, the materials used are supplied by The Himalaya Drug Co., in different strengths and in suspension and powder form. We have prepared suspension in water from Cystone powder using Carboxy-methyl-cellulose as the suspending agent. (Carboxymethyl-cellulose was also supplied by The Himalaya Drug Co.). 1. Diuretic activity of Cystone Healthy albino rats weighing between 100 g to 150 g were selected for the study. Aston s (1959) method was followed. According to this method for the test, a selection of uniform rats was made. The selection depended upon their response to primary hydration. Subsequently, the rats chosen were given secondary hydration of saline followed by gastric intubation of Cystone in different strengths and doses. The diuretic and saluretic responses were measured for a period for five hours.

2 For the actual experiment 72 rats were selected. Cystone was administered orally to each group, in doses of 100 mg, 200 mg, 300 mg, 400 mg and 500 mg per 100 g of body weight. For each dose, three groups consisting of 3 pairs of rats (six rats) to each group were allotted. A group of six rats was taken as control. The diuresis was observed for five hours. The albino rats were deprived of food for eighteen hours and water was given ad lib. For the primary hydration test, the animals were administered orally 0.1 per cent saline in an amount equivalent to 2 per cent of their respective body weights and placed in specially prepared metabolic cages. The volume of urine excreted over the next two hours was measured. Those rats whose urinary output was 50% or more than that of the primary dose of saline were selected for the test. After the primary hydration test, the selected rats were given 1% saline, orally equivalent to 4% of their respective body weights, followed by oral administration of Cystone in different doses. A group of six rats was treated as control and they were hydrated with the same amount of saline. One per cent Carboxy-methylcellulose aqueous solution was administered to the controls. The animals were again placed in the metabolic cages and the urinary output was recorded every hour over a period of five hours and expressed cumulatively as milliliter excreted per kilogram of body weight. 2. Saluretic effect of Cystone (Sodium, Potassium and Calcium Estimation) The urinary output was also utilized for chloride estimation of the above electrolytes. The total amount of sodium, potassium and calcium for five hours were determined by Lange Flame Photometer and expressed as Milliequivalents (MEq) excreted per litre of urine excretion. 3. Estimation of total protein for any colloidal changes with Cystone Four groups of albino rats (each group containing ten rats) were taken. Group A was taken as control to which 4 ml of distilled water was administered orally. After one hour, the rats were sacrificed and their blood collected in centrifuge tubes. Similarly, in the other three groups, Cystone suspension was administered orally in doses of 500 mg, 300 mg and 100 mg in equal value of 4 ml solution. 1 mg of serum was taken and to it 9 mg of 1% NaCl was added and mixed. From this 1 mg of the diluted serum was transferred to a 15 ml centrifuge tube and 4 ml of water and 1 ml of 20% T.C.A. were added and mixed thoroughly. After 15 minutes centrifuge the liquids were poured off, draining off as much as possible. To the precipitated protein 1 mg of 10% NaOH was added and heated on boiling water bath for 30 minutes, with a filter funnel in the mouth of the tube to lessen evaporation. After removing from the water bath, 7 ml of water, 1 mg of phenol reagent and 3 ml of a saturated solution of sodium carbonate were added in that order, and mixed by inversion.

3 Standard: To 2 ml of 0.02% Tyrosine, 1 ml of 10% NaOH and 5 ml of water were used in a centrifuge tube. Then 1 mg of phenol reagent and 3 ml of a saturated solution of sodium carbonate were added and mixed by inversion. Reading was taken after 30 minutes standing by the calorimetric method of protein estimation. 4. Effect of Cystone on smooth muscles Studies were done on isolated loops of rabbits and guinea pigs intestine in Dales isolated organ bath by the method of Transdelenburg as modified by Magnus. The effects of Cystone on the normal movements as well as on the spasm induced by Cystone were studied. 5. Toxicity studies on Cystone Acute toxicity and chronic toxicity studies were carried out on rats. RESULTS 1. Diuretic Effect To a number of 72 rats, Cystone was administered orally to each group in the dose of 100, 200, 300, 400 and 500 mg per 100 g of body weight. In Table 1, the dose-response to Cystone as a diuretic are shown. Table 1: Diuretic effect of Cystone Amount of urine excreted in ml per kilogram in 5 hours 100 mg 19 ml 200 mg 19 ml 300 mg 20 ml 400 mg 30 ml 500 mg 39 ml 18 ml Upto the dose of 200 mg per 100 g of body weight, there was no significant difference in the volume of urine excretion as compared with controls. At dose level of 300 mg per 100 g of body weight, there was a slight increase in the volume of urine excretion, than in the controls. In doses of 400 mg and 500 mg there was significant increase in the urine volume. The effective dose for diuresis in rats lies between 400 to 500 mg per 100 g of body weight of rats.

4 2. Saluretic effect (a) Excretion of sodium: No significant change in the excretion of sodium upto the dose of 200 mg per 100 g of body weight was observed. In dose of 300 mg per 100 g of body weight, there was a slight increase in sodium excretion than in the controls. In doses of 400 mg and 500 mg, there was significant increase in the sodium excretion (Table 2). (b) Excretion of potassium: In doses of 400 mg to 500 mg, there was a significant increase in the potassium excretion, but the excretion of potassium was relatively less than the sodium excretion (Table 3). Table 2: Saluretic effect of Cystone meq/l 100 mg 50 meq/l 200 mg 50 meq/l 300 mg 100 meq/l 400 mg 180 meq/l 500 mg 190 meq/l 40 meq/l Table 3: Excretion of Potassium meq/l 100 mg 10 meq/l 200 mg 20 meq/l 300 mg 20 meq/l 400 mg 30 meq/l 500 mg 40 meq/l 10 meq/l (c) Excretion of calcium: There was a significant increase in the excretion of calcium in doses of 400 to 500 mg per 100 g of body weight (Table 4). Table 4: Excretion of Calcium meq/l 100 mg 0.3 meq/l 200 mg 0.3 meq/l 300 mg 0.5 meq/l 400 mg 0.9 meq/l 500 mg 1.0 meq/l 0.3 meq/l It was found that in doses of 300 and 100 mg of Cystone, the amount of protein present in g per 100 ml of blood was lower than in control, which was not significant. In 500 mg dose of Cystone, the amount of protein present in g per 100 ml of blood was found significantly lower than in control, as shown in Table 5.

5 No. of Table 5: Total protein present in g per 100 ml of blood of albino rats 500 mg of Cystone 300 mg 100 mg No. of No. of No. of DISCUSSION Cystone was administered in different doses to laboratory animals (rats) and it was found that it possesses a good diuretic activity. This was more significant in 400 mg and 500 mg doses per 100 g of body weight in rats. In doses of 200 mg per 100 g of body weight, there was no significant difference in the volume of urine excretion as compared with control volume. How Cystone exerts its diuretic effect is still not known. Its spasmolytic activities on smooth muscles of the bladder, ureter and kidney might have some effects in the production of diuresis along with other mechanisms. In doses of 400 mg and 500 mg there was a significant increase in sodium excretion. Similarly in the same doses, potassium excretion also occurred. But comparatively the excretion of potassium was relatively less than the sodium excretion. Calcium excretion also showed a significant increase at doses of 400 to 500 mg per 100 g of body weight. Thus Cystone caused increased excretion of electrolytes (i.e. sodium, potassium and calcium) in urine and as such possesses a good diuretic activity, which can be well compared with known potent diuretics. Cystone exhibited a good spasmolytic activity on the smooth muscles of the rabbits and guinea pigs. However, its effect on acetylcholine, histamine and barium-induced spasm of the smooth muscles is not so marked. In rats administered intraperitoneally in very high doses up to 500 mg per 100 g of body weight, Cystone did not produce any toxic effects. No rats died within 24 hours, nor any changes were observed in heart rate and respiration. All these observations confirm the previous reports of its nontoxic effects. About the study of crystalloid-colloid balance, we studied the effects of Cystone on serum proteins of normal animals, taking a group as control and to the other groups administering Cystone in doses of 100 mg, 300 mg and 500 mg respectively. It was observed that at dose level of 500 mg, Cystone caused a slight lowering of plasma proteins in rats. From these findings it may be suggested that Cystone lowers the crystalloid level in the body (through urine) and this lowering effect might have some influence in correcting the crystalloid-colloid balance. Although it is too early to predict its exact mechanism of action it may be suggested that Cystone acts on the binding materials of the calculus and allows its slow disintegration. On human studies sufficient cases have been reported until now.

6 CONCLUSION Cystone has an active and potent role in relieving cases of early urolithiasis. Although the aetiology of formation of urinary calculus is still unknown, it may be remarked that in early cases, Cystone therapy will be more efficacious and beneficial than surgical interference. SUMMARY 1. Cystone, a product of The Himalaya Drug Co., has been investigated pharmacologically on animals. 2. The pharmacological studies on animals showed that it possesses a good diuretic activity and spasmolytic effect on the smooth muscles of animals. 3. It has no toxic effects on animals. 4. It causes excretion of electrolytes in the urine of animals and significantly lowers the serum proteins in animals. Although the exact mechanism involved in correcting the crystalloid-colloid balance is not known, it may act on the binding material of the calculus and allow its disintegration. It has a potent role clinically in relieving early cases of urolithiasis. It may be suggested that Cystone may be more efficacious and beneficial than surgical interference in the early cases of urolithiasis. ACKNOWLEDGEMENT The authors express their deep sense of sincere gratitude to the Principal, Gauhati Medical College and the Medical Authorities for allowing the facilities to carry out the research work and The Himalaya Drug Co., for kindly offering their assistance in the investigations. They are also indebted to the Department of Bio-chemistry, Gauhati Medical College and Department of Geology, Gauhati University for their kind help and assistance for carrying out the biochemical investigations.