Evaluation of In Vitro Antiurolithiatic Activity of Vigna radiata

 

Spandana. K*, Shivani. M, Himabindhu. J, Ramanjaneyulu. K

Department of Pharmacognosy, Vishnu Institute of Pharmaceutical Education and Research,

Narsapur, Medak, Telangana, India.

*Corresponding Author E-mail: spandana2503.1998@gmail.com

 

ABSTRACT:

The present study was undertaken to evaluate the in vitro antiurolithiatic activity of the medicinal plantvigna radiata aqueous extract.It showed its maximum efficiency in the dissolution of calcium oxalate crystals.. Our result have clearly indicated that the Aqueousextract of seeds ofvignaradiata were quite promising for further studies in this regard. In this study Neeri was used as standard drug.

 

KEYWORDS: In vitro antiurolithiatic activity, Aqueous extract, urolithiasis, vigna radiata, Neeri.

 

 


INTRODUCTION:

Plants provide food, raw materials for medicine and various other requirements for the very existence of life from the origin of human beings1. Urinary stone formation affects 10–12% of the population in industrialised countries. From epidemiological data, calcium oxalate (CaOx) is the most common component of the calculi2. The formation of such concretions involves several physicochemical events, e.g. nucleation, growth and aggregation, but the mechanism(s) of these processes remain incompletely understood3. Furthermore, although some drugs used to prevent the disease have some positive effects, they are not effective in all patients and often have adverse effects that compromise their use in longterm medical treatment.4

 

Kidney stone disease is a multi-factorial disorder resulting from the combined influence of epidemiological, biochemical and genetic risk factors3. Urolithiasis is considered as the third most common affliction of the urinary tract. It refers to the solid non-metallic minerals in the urinary tract.

 

 

 

It is a complex process that is a consequence of an imbalance between promoters and inhibitors in the kidney. The formation of kidney stones involves several phytochemical events beginning with crystal nucleation, aggregation and end with retention within the urinary tract. Among the several types of kidney stones, the most common are calcium oxalate stones representing up to 80% of the analyzed stones4.

 

Calcium containing stones may be in the form of pure calcium oxalate(50%) or calcium phosphate(5%) and a mixture of both(45%) followed by magnesium phosphate (15-20%), uric 5 acid(10%) and cystine (1%)5. Many patients still undergo surgery to remove the stones; thus in Morocco, as in many countries, most patients (≈70%) use medicinal plants as an alternative therapy for many diseases, including lithiasis. The aerial parts of Herniariahirsuta, widely distributed in the Mediterranean area, is used in folk medicine as a diuretic and to treat kidney stones6Though technological advancements have made dramatic improvement in the removal of urinary stones still some of the drawbacks of these methods exists which includes their being too costly for a common man and recurrence of stone formation along with a number of other side effects.7

 

Management of stone disease depends on the size and location of the stones. Stones larger than 5 mm or stones that fail to pass through should be treated by some interventional procedures such as extracorporeal shock wave lithotripsy (ESWL), ureteroscopy (URS), or percutaneous nephrolithotomy (PNL)8

 

The seeds and sprouts of mung beans are also widely used as a fresh salad vegetable or common food in India, Bangladesh, South East Asia, and western countries9. The energy offered by mung beans and sprouts is lower than that of other cereals, which is beneficial for individuals with obesity and diabetes10. Regular consumption of mung beans could regulate the flora of enterobacteria, decrease the absorption of toxic substances, reduce the risk of hypercholesterolemia and coronary heart disease, and prevent cancer11

 

2. MATERIALS AND METHODS:

PLANT MATERIAL:

The seeds of Vigna radiatawas collected in the month of march 2018 from sangareddy town, sangareddy dist. of Telangana, India.The seeds were washed with tap water and dried under shade.

 

PREPARATION OF PLANT EXTRACT:

The seeds were shade dried and powdered. The crude plant extract was prepared by Soxhlet extraction method. 50g of powdered plant material was extracted with 500ml of water. The process of extraction was carried out up to 6 cycles, till the solvent in siphon tube of an extractor became colorless. The extract was filtered, and evaporated to dryness using rotary evaporator. Further the dried extract was maintained in a refrigerator at 4ºC for further antiurolithiatic activity.

 

CHEMICALS USED:

Neeri, Sodium oxalate, Tris buffer, calcium chloride, Potassiumpermanganate (KMnO4), Sulphuric acid (H2SO4).

 

Investigation of in vitro antiurolithiatic activity test by titrimetry:

The experimental kidney stones of calcium oxalate (CaOx) were prepared in the laboratory by taking equimolar solution of calcium chloride dehydrate in distilled water and sodium oxalate in 10 ml of 2N H2SO4. Both were allowed to react in sufficient quantity of distilled water in a beaker, the resulting precipitate was calcium oxalate. The precipitate was freed from traces of sulphuric acid by ammonia solution, washed with distilled water and dried at 600C. The dissolution percentage of calcium oxalate was evaluated by taking exactly 1 mg of calcium oxalate and 10 mg of the extract, packed it together in semi permeable membrane of egg as shown in the model designed given below .This was allowed to suspend in a conical flask containing 100 ml of 0.1M Tris buffer. First group served as blank containing only1 mg of calcium oxalate. The second group served as positive control containing 1mg of calcium oxalate and along with the 10mg standard drug, i.e. Neeri. The 3rd group along with 1 mg of calcium oxalate contain aqueous extract. The conical flasks of all groups were kept in an incubator preheated to 37oC for 2 h. Remove the contents of semi permeable membranes from each group into separate test tubes, add 2 ml of 1Nsulphuricacid to each test tube and titrated with 0.9494 N KMnO4 till a light pink colour end point obtained. The amount of remaining undissolved calcium oxalate is substracted from the total quantity used in the experiment in the beginning to know the total quantity of dissolved calcium oxalate by various solvent extracts12.

 

3. RESULTS AND DISCUSSION:

Drug therapy has developed in response to population health care13 needs. There are many crucial areas in medicine such as liver diseases, arthritis, old age related problems, certain viral infections and cancer where the conventional medicine is devoid of satisfactory treatment. These are among the promising areas of research and development of medicines from the vast highly potential plant resources. Plants are also attractive sources for the development of novel and very effective and safe therapeutic agents against kidney procumbens. Herbal medicines are also in great demand in the developed world for primary health care because of their efficacy, safety and lesser side effects14. Unlike allopathic medicines which target is only one aspect of urolithiatic pathophysiology, most of plant based therapy have been shown to be effective at different stages of stone pathophysiology15. About 80% of the world populations rely on the use of traditional medicine which is predominantly based on plant materials16. Plant based drug discovery programmes continue to provide an important source of new drug leads17. Lithiasis (stone formation) is an important cause for acute and chronic renal failure, includes both nephrolithiasis (stone formation in kidney) and urolithiasis (stone formation in ureter or bladder or both). Among the various kinds of stones identified, calcium stones occur mainly in Men, while phosphate stones formation is more in women18.

 

This study evaluates the antiurolithiatic activity of Aqueous extract of Vigna radiata. The highest percentage i.e. 98.1% of calcium oxalate {CaOx} dissolution was observed in Aqueous extract. Aqueous extract of Vigna radiata was found to be more effective in dissolution of calcium oxalate than standard drug Neeri. This study has given primary evidence for Vigna radiata as the plant which possess lithotriptic property. This in vitro study has given lead data and shown that Aqueous extract is quite promising for further studies in this regard.

 

Table 1: Shows % dissolution of calcium oxalate (CaOx) by Vigna radiataseeds extracts.

 

% of dissolution of calcium oxalate

S.No

GROUPS

Vigna radiate

1.

Blank

0

2.

Positive Control

81

3.

Aqueous extract

98.1

 

Figure 1(a): Decalcification of egg shell in 10% Acetic acid overnight.

 

 

Figure 1(b): Decalcified Eggs

 

 

Figure 1(c): Egg membrane along with the contents suspended into the 0.1 M Tris buffer.

Figure 1: In vitro experimental model setup to evaluate antiurolithiatic activity.

 

4. CONCLUSION:

In vitro urolithiasis has been performed on the selected plant Vigna radiata by using the standard drug, Neeri. The work was performed by using in vitro antiurolithiatic model for calculating percentage dissolution of kidney stone. Aqueousseed extracts of Vigna radiatashows highest dissolution than standard drug Neeri. This study has given primary evidence for vigna radiate as the plant which possess antiurolithiatic property.

 

 

5. ACKNOWLEDGEMENT:

We sincerely thankful to our principal Dr. A. Ramesh and staff members, Director and chairman of our college Vishnu Institute of Pharmaceutical Education and Research (VIPER) for supporting us.

 

6. REFERENCES:

1.       Sumayyasikandari and Prathima Mathad(2015). In vitro antiurolithiatic activity of Butea monosperma Lam. and Nigella sativa Linn.seeds.Ukaaz-Annals of Phytomedicine,4(1):105-107.

2.       Finlayson B. Symposium in renal lithiasis. Renal lithiasis in review Urolclin North Am 1974; 1 181 212

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6.       Bellakhdar J, Claisse R, Fleurentin J, Younos C. Repertory of standard herbal drugs in the Moroccan pharmacopoeia. J Ethnopharmacol 1991; 35: 123 43

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8.       Coll DM, Varanelli MJ, Smith RC. Relationship of spontaneous passage of ureteral calculi to stone size and location as revealed by unenhanced helical CT. AJR Am J Roentgenol. 2002;178:101–103

9.       Fery RL: The cowpea: production, utilization, and research in the United States. Horticultural Reviews. 1990, 12: 197-222.

10.     Zheng JX: Functional foods-second volume. 1999, Beijing: China Light Industry Press

11.     Kruawan K, Tongyonk L, Kangsadalampai K: Antimutagenic and co-mutagenic activities of some legume seeds and their seed coats. J Med Plants Res. 2012, 6 (22): 3845-3851.

12.     Unnate Atodriya; Roshni Baard; Siddi Upadhya and Umesh Upadhyay (2013). Antiurolithiatic activity of Dolichosbiflorus seeds. Journalof Pharmacognosy and Phytochemistry, 2(2): 209-213.

13.     Michael Dickson and Jean paulGagnon (2004). Key factors in the rising cost of new drug discovery and development. Nature Reviews Drug Discovery, 3:417-429.

14.     Kamboj V.P (2000). Herbal medicine.Current Science Association (JSTOR), 78(1): 35- 39.

15.     Archana R.Dhole, Vikas R. Dhole, Chandrakant S. Magdum, Shreenivas Mohite (2013). Herbal Therapy for Urolithiasis: A Brief Review. Research Journal of Pharmacology and Pharmacodynamics.5(1):6-11.

16.     Subramoniam. A, P. Pushpangadan (1999). Development of phytomedicines for liver disease. Indian Journal of Pharmacology. 31(3):166-175.

17.     Sanjay M,Jachak and Arvind saklani (2007).Challenges and opportunities in drug discovery from plants.Current Science Association (JSTOR), 92(9):1251-1257.

18.     Ramachandran. S, Vijayakumar. T.M, Saisandeep.V, Ramsai. K and Dhanaraju. M.D (2011). Antilithiatic Activity of Poly Herbal Extracts on Ethylene glycol-Induced Lithiasis in Rats.European Journal of Biological Sciences, 3(2):36-39.

 

 

 

 

 

 

Received on 03.08.2018          Modified on 31.08.2018

Accepted on 27.09.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2018; 11(12): 5455-5457.

DOI: 10.5958/0974-360X.2018.00994.0