Calculi

Byline: M. Alvin Jose, Ibrahim, S. Janardhanan

Sir,

Nephrolithiasis is worldwide in distribution and affecting 2% of the world population.[1] In order to find a herbal remedy for this disease, the present study was undertaken to evaluate the antilithiotic activity of the concentrated fresh juice of the leaves of Plectranthus amboinicus Lour. It is popularly known in English as Indian borage, syn. Coleus amboinicus (Family: Lamiaceae). Indians use it widely for various illnesses, including kidney stone.[2] It is scientifically evaluated for its antibacterial[3] and antioxidant activity.[4]

The fresh plants of P. amboinicus were collected from the herbal garden, Avinasilingam Deemed University, Coimbatore and authenticated at the botany department. About 500 g of fresh leaves were cut to small pieces and fresh juice was prepared adding water (30 ml), with the help of mixer. The fresh juice was filtered and concentrated to a dry mass by vacuum distillation; black dry residue was obtained (16 g). The LD50 was done using OECD guideline for testing of chemicals revised draft guideline 423. The one tenth of the LD50 500 mg/kg was chosen as a dose for the further study.

Adult male albino rats of the Wistar strain, weighing between 150-200 g were used for this study. Animals were acclimatized and maintained at 24o C[+ or -]2o C, 70% RH and12h/12h light and dark cycle throughout the study. They were fed with standard pellet diet (Hindustan liver limited, Bangalore, India) and water ad libitum . The CPCSEA and the local ethical committee approved the studies. The animals were divided into three groups G1, G2, and G3 of six animals each. Following the method of Tamilselvan,[5] nephrolithiasis was induced. Group 1(G1: control) was fed with ordinary drinking water. Group 2(G2: lithiotic control) were fed with 1% ethylene glycolated water for 35 days. Group 3(G3: test group) were fed with 1% ethylene glycolated water and were simultaneously administered, by gastric tube, 500 mg/kg of concentrated fresh juice of the leaves of the Plectranthus amboinicus Lour for 35 days. On the 34th day all the animals were placed in metabolic cages and urine was collected for 24 hours and urine was analyzed for the presence of calcium, oxalate and total protein. Calcium,[6] and total protein[7] were estimated by an auto analyzer (Logotech s.r.l, tecno-168). Oxalates were analyzed by the method of Hodgkinson and Williams.[8] On Day 35 the animals were sacrificed, the kidneys were excised, washed with cold saline, fixed in 10% formalin solution and subjected to histopathological studies.

The results are expressed as mean [+ or -] SEM. The data were subjected to one-way ANOVA followed by Turkey-Kramer multiple comparison tests. P values < 0.05 were considered as significant.

Histopathological studies clearly revealed that the tissue samples from the control group (G1) shows tubules with single epithelial lining along the margin and were of normal size. In G2 (lithiotic control), all the tubules showed the presence of crystals, there was marked dilatation of the tubules and total degeneration of the epithelial lining with infiltration of inflammatory cells into the interstitial space. In G3 (test Group) the specimen showed characters similar to the control group. Urine analysis showed a significant elevation of calcium, oxalates and total proteins level in the lithiotic control group (G2), when compared to normal control. The test group (G3), showed a significant reduction in all the parameters almost comparable with normal control. The urine and histopathological results clearly revealed the antilithiotic activity of P. amboinicus , particularly of calcium oxalate origin. Further research is needed to explore the exact active principle(s) responsible for the antilithiotic activity and the mechanism of action.

References

1. Mohan H. Text book of pathology. 4th Ed. New Delhi: Jaypee Brother's Medical Publication (P) Ltd; 2000.

2. Orient Longman. Indian Medicinal Plants. Vol. 4. Madras. Microprint; 1995.

3. Timer CE, Marzini ME, Fernandez A, Gonzalez ML. Physiochemical assessment of the essential oil from the leaves of Plectranthus amboinicus Lour. Sprena growing in Cuba. Rev Cubana Farm 1992;251:63-8.

4. Padma PR, Bhuvaneswari V, Chelvi SK.The activities of enzymic antioxidant in selected green leaves. Indian J Nutrition Dietetics 1998;35:1-3.

5. Tamilselvan S, Raymond L, Khan SR. Lipid peroxidation in ethylene glycol induced hyperoxaluria and calcium oxalate nephrolithiasis. J Urol 1997;157:1059-63.

6. Sarkar RC, Chauhan SP. A new method for determining micro quantities of calcium in biological materials. Anal Biochem 1967;20:155-6.

7. Goruall AG, Bardawill CJ. Determination of serum proteins by means of biuret reaction. J Biol Chem 1949;177:751-66.

8. Hodgkinson A, Williams A. An improved colorimetric procedure for urine oxalate. Clin Chem 1972.

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