Journal of Alternative Complementary & Integrative Medicine Category: Medicine Type: Research Article
An Aqueous Extract of the Leaves of Ficus religiosa Inhibits the Growth of Urinary Calcium Hydrogen Phosphate Dihydrate Crystals-An In-Vitro Study
- Baskaraboopathy A1, Rajaram MG1, Elizabeth AR1, Eevera T1, Jayakumar T2*
- 1 Department Of Biotechnology, Periyar Maniammai University, Tamil Nadu, India
- 2 Department Of Pharmacology, Graduate Institute Of Medical Sciences, Taipei Medical University, 250 Wu-Hsing Street, Taipei 110, Taipei, Taiwan, Province Of China
*Corresponding Author:Jayakumar T
Department Of Pharmacology, Graduate Institute Of Medical Sciences, Taipei Medical University, 250 Wu-Hsing Street, Taipei 110, Taipei, Taiwan, Province Of China
Tel:+886 227361661, 3205
Received Date: Jul 18, 2017 Accepted Date: Oct 25, 2017 Published Date: Nov 09, 2017
Methods: CHPD crystals were grown as a urinary type using single diffusion gel technique in silica hydro-gels. An extract (50, 75 and 100%) of the leaves of F. religiosa was added over to CHPD crystals growth for 12 days to evaluate its inhibitory effect. Thermogravimetric (TGA) and powder X-ray diffraction analyses were performed to characterize the cultured CHPD crystals.
Results: A 75 and 100% aqueous extracts of the leaves of F. religiosa was significantly (P<0.05 and 0.01, respectively) reduced CHPD crystals growth and the inhibition rate was almost >50%. Whereas, a moderate inhibition (P<0.05) of CHPD crystal growth was found in 75% of aqueous leaf extracts of F. religiosa. However, 50% of aqueous leaf extracts of F. religiosa exhibit no inhibition on the CHPD crystal growth.
Conclusion: These results indicate that the higher concentration of the leaf extracts of F. religiosa is highly responsible for the inhibition of CHPD crystal growth. This study may suggest that F. religiosa can be used as therapeutic agent for the treatment of urinary calculi or their prevention.
The development of crystals in the gel is the simplest techniques under ambient environments which is appropriate for the crystal growth of compounds sparingly soluble and which decompose at low temperatures. India is expected to have urinary stones at about 12% of the population and out of that about 50% of cases encounter loss of one or both kidneys. It has been shown that upper as well as lower urinary tract stones arise habitually, but in India the incidence shows wide variation on the regional basis . Therefore, it is essential to find for an alternative means such as medicinal plants or phytotherapy . Data from in vitro, in vivo studies and clinical trials reveal that phytotherapeutic agents could be useful as either an alternative or an adjunctive therapy in the management of urolithiasis .
Ficus religiosa L. (Lauraceae), is widely branched with long-tipped, leathery heart shaped leaves and purple fruits growing in pairs. It has mythological, religious and medicinal importance in Indian culture since ancient times [7,8]. This plant has long been used in traditional medicine for various disorders and its different parts have been used medicinally in various forms as well as in combination with other herbs. This plant has been shown to have numerous biological activities such as wound healing, antibacterial, anticonvulsant, antidiabetic, antiinflammatory and acetyl cholinesterase inhibitory activity [8-13]. The acetone extract of F. religiosa leaves has been shown to induce apoptosis in breast cancer cell lines . Choudhari et al., have reported the antioxidant and cytotoxic activity of F. religiosa bark against cervical cancer cells . A recent study has reported the anti-ulcer activity of the ethanolic extract of F. religiosa leaf . Antitumor activity of biosynthesized silver nanoparticles using F. religiosa as a nanofactory in Dalton’s Ascites Lymphoma (DAL) induced mice has recently been reported by . In the present study, for the first time we aimed at investigating the efficacy of F. religiosa on CHPD crystals growth in vitro.
MATERIALS AND METHODS
Growth of CHPD crystals
|SMS gel density gm/cm3||Orthophosphoric acid concentration||Gel + H3PO4pH||Gel settingTime (hr)||Concentration of SupernatantCaCl2(M)||Liesegangring formation(hr)||Growthperiod (days)||Types of crystals||Harvested crystals size (cm)|
|1.04||0.5 N||6.4||34||1||12||60||Single crystals||1.7 - 2.0|
|1 N||7.3||24||1||24||35||Rod, stars shaped||2.3 - 3|
Collection Ficus religiosa L. (Lauraceae) and its extract preparation
Figure 1A: The leaves of Ficus religiosa L.
F. religiosa extract on the growth of CHPD crystals
Thermogravimetric analysis of CHPD crystals
Powder X-ray diffraction analysis of CHPD crystals
Figure 2A: Growth of CHPD crystals at the end of 12th days.
|Shape||Needle or star shaped|
|Length||2.3 - 3 cm|
|Breadth||1 - 2 mm|
|Thermo gravimetric analysis|
|Points||Temperature||Presence of CHPD crystal|
The X-ray powder diffraction of the cultured CHPD crystals (Table 4) was matched with the organic database-using computer and the results were consistent with CHPD X-ray powder diffract gram (Figure 3A) produced during the x-ray scans. The peaks positions represent where the x-ray beam has been diffracted by the CHPD crystals. The set of d spacing (the distance between the adjacent planes of atoms), which represents the unique “finger prints” of the crystals can easily calculated from the 2-theta (2q) values. The uses of the degrees 2-theta in depicting x-ray powder diffraction scans is a matter convention and can easily be related peak to the JC-PDF (Joint Committee for Powder x-ray Diffraction File). The comparing the measured diffract grams with JC-PDF has identified constituents present in the CHPD crystals.
Figure 3A: Powder X-ray diffractogram of CHPD crystals.
|Position||Height (cm)||FWHM (02Th.)||d-spacing||Relative|
It has been reported that oxalate plays an important role in stone formation and has about 15-fold greater effect than urinary calcium [24,25]. In the present study, CHPD is formed competently in an in vitro gel system. Joshi et al., have reported the inhibitory effects of litholytic medicinal plants Tribulus terrestris L. (Zygophyllaceae) and Bergenia ligulata L. (Saxifragaceae) on the growth of CHPD crystals . They have described that calcium chloride containing supernatant solution produced Liesegang rings in the gel and needle, platelet and star-shape CHPD crystals which are grown within the rings. The calcium chloride in the supernatant solutions was modified the diffusion process and thus the periodic precipitation and the number of Liesegang rings reduced by the addition of aqueous extracts of T. terrestris and B. ligulata . They have also noted the maximum length of the crystals reduced due to inhibition produced by the addition of aqueous extracts of B. ligulata and T. terrestris. Several lines of evidence indicated that there are various compounds exhibiting inhibitory actions on the growth of urinary stones and crystals, such as tartrates are good inhibitors of stones in natural and artificial urine . In the present study, an extract of F. religiosa was used to study the growth behavior of CHPD crystals. F. religiosa inhibits the growth of CHPD crystals. This can be verified from the results of the formation of CHPD crystals.
Numerous inhibitors have been identified in urine for the calcium phosphate and calcium oxalate crystal systems. Magnesium, citrate, pyrophosphate and nephrocalcein are the inhibitors in the calcium phosphate crystal system . The in vitro formation of urinary stones and the generation of spherulites of calcium phosphate in gels, as well as the overgrowth with calcium oxalate using a new flow model of crystallization have been studied by Achilles et al. . Ethanolic extract of T. terrestris fruits were found to exhibited protection against uroliths induced by glass bead implantation in rats . In another study, an administration of a drug containing T. terrestris to sodium glycolate fed rats produced a significant decrease in urinary oxalate excretion and a significant increase in urinary glyoxylate excretion . Recently various herbal plants such as Flos carthami, Costus igneus, Tribulus terrestris and Scoparia dulcishave successfully proved as prophylactic and curative medicine for urolithiasis [32-35]. Another recent in vitro study showed that leaf extracts of I. eriocarpa possesses potent antiurolithiatic activity . So the results of present study corroborate findings with the results of the above investigations.
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Citation:Baskaraboopathy A, Rajaram MG, Elizabeth AR, Eevera T, Jayakumar T (2017) An Aqueous Extract of the Leaves of Ficus religiosa Inhibits the Growth of Urinary Calcium Hydrogen Phosphate Dihydrate Crystals-An In-Vitro Study. J Altern Complement Integr Med 3: 041.
Copyright: © 2017 Baskaraboopathy A, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.