Antiurolithiatic effects of Cassia fistula Lin. fruit extracts on ethylene glycol-induced nephrolithiasis in rats.

Urinary stones are a growing disease that results from pathological biomineralization. Cassia fistula Lin. is traditionally used to treat urinary stones. However, no scientific evidence is available to prove its antilithiatic effect. This study evaluates the antilithiatic potential of aqueous and ethanolic extract of Cassia fistula Lin. fruit (Cff) against calcium oxalate kidney stones. Forty-two male Wistar rats were divided into seven groups (n = 6/group): Group I (control), Group II (rats treated with ethylene glycol and ammonium chloride developed nephrolithiasis after 28 days), Group III (lithiatic rats receiving distilled water for 30 days), Group IV and V (lithiatic rats receiving aqueous extract of Cff at doses of 1 and 100 mg/kg body weight for 30 days, respectively) and Group VI and VII (lithiatic rats receiving ethanolic extract of Cff at doses of 1 and 100 mg/kg body weight for 30 days, respectively). Some parameters of urine and serum, and also renal oxidative stress and histopathology were used to determine the antilithiatic effect of aqueous and ethanolic extract of Cff. Therefore, the types of extracts of Cff improved abnormal levels of urine, serum, and renal oxidative stress and histopathology parameters. This antilithiatic effect of aqueous and ethanolic extracts of Cff, can be attributed to the anti-crystallization and antioxidant properties of the extracts and the ability to improve urine and serum biochemistry. RESEARCH HIGHLIGHTS: Ethylene glycol and ammonium chloride-induced urolithiasis, aggregation of calcium oxalate deposits, increase of some urinary and serum parameters, relative kidney weight, kidney size and MDA activity, decrease of some urinary parameters, relative body weight and SOD activity. Aqueous and ethanolic extracts of Cassia fistula Lin. lead to the treatment of urolithic rats by decreasing levels of urinary oxalate, phosphate, urea, serum urea, uric acid, creatinine, calcium, phosphate, MDA, kidney weight and kidney size, increasing levels of urinary calcium, creatinine, magnesium, citrate, body weight and SOD activity in the kidney, eliminating CaOx deposition (esp. ethanolic extract).

The inhibitory effects of epigallocatechin-3-gallate on calcium oxalate monohydrate crystal growth, aggregation and crystal-cell adhesion.

Epigallocatechin-3-gallate (EGCG), a predominant phytochemical in tea plant, has been reported to prevent kidney stone formation but with vague mechanism. We investigated modulatory effects of EGCG (at 0.1-100 µM) on calcium oxalate monohydrate (COM) crystals at various stages of kidney stone development. EGCG significantly increased crystal size (at 1-100 µM), but decreased crystal number (at 10-100 µM), resulting in unchanged crystal mass and volume. Interestingly, EGCG at 10-100 µM caused morphological change of the crystals from typical monoclinic prismatic to coffee-bean-like shape, which represented atypical/aberrant form of COM as confirmed by attenuated total reflection - Fourier transform infrared (ATR-FTIR) spectroscopy. EGCG at all concentrations significantly inhibited crystal growth in a concentration-dependent manner. However, only 100 µM and 10-100 µM of EGCG significantly inhibited crystal aggregation and crystal-cell adhesion, respectively. Immunofluorescence staining (without permeabilization) revealed that surface expression of heat shock protein 90 (HSP90) (a COM crystal receptor) on MDCK renal cells was significantly decreased by 10 µM EGCG, whereas other surface COM receptors (annexin A1, annexin A2, enolase 1 and ezrin) remained unchanged. Immunoblotting showed that 10 µM EGCG did not alter total level of HSP90 in MDCK cells, implicating that its decreased surface expression was due to translocation. Our data provide a piece of evidence explaining mechanism underlying the anti-lithiatic property of EGCG by inhibition of COM crystal growth, aggregation and crystal-cell adhesion via reduced surface expression of HSP90, which is an important COM crystal receptor.

Proteomic and computational analyses followed by functional validation of protective effects of trigonelline against calcium oxalate-induced renal cell deteriorations.

Trigonelline is a phytoalkaloid commonly found in green and roasted coffee beans. It is also found in decaffeinated coffee. Previous report has shown that extract from trigonelline-rich plant exhibits anti-lithiatic effects in a nephrolithiatic rat model. Nevertheless, cellular mechanisms underlying the anti-lithiatic properties of trigonelline remain hazy. Herein, we used nanoLC-ESI-Qq-TOF MS/MS and MaxQuant-based quantitative proteomics to identify trigonelline-induced changes in protein expression in MDCK renal cells. From a total of 1006 and 1011 proteins identified from control and trigonelline-treated cells, respectively, levels of 62 (23 upregulated and 39 downregulated) proteins were significantly changed by trigonelline. Functional enrichment and reactome pathway analyses suggested that these 62 altered proteins were related to stress response, cell cycle and cell polarity. Functional validation by corresponding experimental assays revealed that trigonelline prevented calcium oxalate monohydrate crystal-induced renal cell deteriorations by inhibiting crystal-induced overproduction of intracellular reactive oxygen species, G0/G1 to G2/M cell cycle shift, tight junction disruption, and epithelial-mesenchymal transition. These findings provide cellular mechanisms and convincing evidence for the renoprotective effects of trigonelline, particularly in kidney stone prevention.

Phytochemical characterisation and in vivo antilithiatic activity of the stems of Caesalpinia bahamensis (Brasilete).

The aim of this work was to identify the main chemical constituents and to evaluate the antilithiatic activity of the aqueous and hydroalcoholic extracts of stems of Caesalpinia bahamensis Lam. Fractionation and isolation of constituents from the hydroalcoholic extract was carried out by flash chromatography and semi-preparative liquid chromatography. The antilithiatic activity of the aqueous and hydroalcoholic extracts was evaluated in Wistar rats, where kidney stones were induced by ethylene glycol and ammonium chloride. Creatinine, calcium, and oxalate levels were evaluated and histological analysis was carried out. The homoisoflavonoids protosappanin B, 10-methyl-protosappanin B and brazilin were isolated and the antilithiatic activity of the aqueous and hydroalcoholic extracts was demonstrated by the reduction of the concentration of calcium and oxalate in urine compared to the lithiasis group. It was corroborated by histological analysis. Brazilin and protosappanin B were proposed as chemical markers for this plant species.

Phytochemical composition and biological screening of two Lophophytum species / Composición fitoquímica y cribado biológico de dos especies de Lophophytum

Lophophytum species are holoparasites that grow on tree roots. The objectives of the work were to explore the chemical composition of the tubers of two Lophophytum species and to analyze the antioxidant, anti-inflammatory and antilithiatic activity of their extracts using in vitro methods. The chemical composition was determined by histochemical, phytochemical and TLC tests. In addition, the profile of phenolic compounds was determined by HPLC-MS. The presence of secondary metabolites of recognized activity was demonstrated. The results of the HPLC-MS/MS allowed the tentative identification of catechin, luteolin and glycosides of eriodictyol, naringenin and luteolin in the extract of Lophophytum leandriand eriodictyol, naringenin, luteolin and their glycosylated derivatives in Lophophytum mirabile. The extracts showed promising antioxidant (DPPH, ABTS and ß-carotene-linoleic acid), anti-inflammatory (inhibition of 5-LOX) and anti-urolytic (by bioautographic TLC) activity. It is noteworthy that these are the first results of the phytochemical composition and biological activity of L. mirabile. However, in vivo studies are required to corroborate these activities.

Las especies de Lophophytumson holoparásitas que crecen en raíces de árboles. Los objetivos del trabajo fueron explorar la composición química del túber de dos especies de Lophophytum y analizar la actividad antioxidante, antiinflamatoria y antilitiásica de sus extractos usando métodos in vitro. La composición química se determinó mediante pruebas histoquímicas, fitoquímicas y por TLC. Además, se determinó el perfil de compuestos fenólicos por HPLC-MS/MS. Se demostró presencia de metabolitos secundarios de reconocida actividad. Los resultados del HPLC-MS/MS permitieron identificar tentativamente catequina, luteolina y glucósidos de eriodictiol, naringenina y luteolina en el extracto de Lophophytum leandriy eriodictiol, naringenina, luteolina y sus derivados glicosilados en Lophophytum mirabile. Los extractos mostraron prometedora actividad antioxidante (DPPH, ABTS y ß-caroteno-ácido linoleico), antiinflammatoria (inhibición de la 5-LOX) y antiurolitiásica (por TLC bioautográfica). Es de destacar que estos son los primeros resultados de composición fitoquímica y actividad biológica de L. mirabile. Sin embargo, se requieren estudios in vivo para corroborar dichas actividades.

Antilithiatic activity of a non-pharmacopoeial Unani formulation in chemically induced urolithiasis in rats.

OBJECTIVES: Parshioshan (Adiantum capillus-veneris L.), Duqu (Peucedanum grande C.B. Clarke), Kaknaj (Physalis alkekengi L.) and Kharekhasak (Tribulus terresteris L.) have been selected for this study as they have been associated with medicinal actions for litholytic activity. METHODS: The experiment was carried out in Sprague Dawley rats divided into seven groups, serving as plain control, disease control, standard control, curative A and B and preventive A and B groups. Animals of plain control received distilled water. Remaining six groups received Ethylene glycol 0.75% and Ammonium chloride 1% by adding in the drinking water for the first three days followed by 0.75% Ethylene glycol for 18 days. From 8th day till 21st day, standard control received Cystone in the dose of 750 mg/kg. Preventive and curative test groups were treated with hydroalcoholic extract of the test drug in the dose of 132 mg/kg and 264 mg/kg from 1st to 21st day and 8th to 21st day of calculi induction. RESULTS: Test drug reduced the number of calcium oxalate crystals in the urine; the level of urinary calcium, creatinine, magnesium, phosphorus, sodium and chloride decreased significantly in standard and test groups. The urine volume increased significantly in all the test groups. The level of serum calcium, urea, phosphorus and creatinine were significantly reduced in all the test groups. CONCLUSIONS: These results indicated that the test drug reduced and prevented the growth of urinary stones. Moreover, the test drug also possessed significant antiurolithiatic activity. However, the protective effect was found more than its curative effect.

Comprehensive nutritional profiling and activity directed identification of lead antioxidant, antilithiatic agent from Macrotyloma uniflorum (Lam.) Verdc.

Macrotyloma uniflorum (Lam.) Verdc. (family Fabaceae) is traditionally important pulse used for kidney stone degradation, cold,cough, asthma and in bronchial issues. It is well reported for the antilithiatic activity and nutritional profiling but no attempt has been made to find out the mechanism and nutrient responsible for the activity in its seed decoction. Therefore, lead high altitude variety responsible for the highest nutrient content and antilithiatic activity was selected for the study. Boiled spray dried extract (M1), lead fractionation (Spray dried residual water fraction: M2), and crude powder extract (M3) were nutritionally profiled using proximate composition analysis, Atomic absorption spectroscopy (minerals and trace elements), Ion exchange chromatography (carbohydrate analysis) and NMR spectroscopy (polar and non polar metabolite analysis). These extracts were also screened for the antilithiatic activity and found significantly important. M1 showed more activity than M3. Further, boiled extract was fractionated with different polarity of solvents (hexane, chloroform, n-butanol, residual water extract). All fractions and reference standard (cystone) were screened for the antilithiatic activity. M2 showed highest activity followed by ethyl acteate while hexane and chloroform did not show any activity in comparison to standards. Thus, lead fraction and market reference standard (cystone) were validated for the presence of nutritional components. Moreover, the selected nutritional components were individually and in combinations screened for the activity which revealed that kidney stone degradation was due to the synergetic action of components than the individual. The results clearly pave a way that Macrotyloma uniflorum is best for combating with malnutrition and kidney stone related problems.

Antiurolithiatic effects of pentacyclic triterpenes: The distance traveled from therapeutic aspects.

Globally, approximately 12% of the population is inflicted by various types of urolithiasis. Standard treatments are available both to avert and treat urolithiasis, but with significant adverse side effects. Pentacyclic triterpenes represent a group of naturally occurring compounds which holds immense potential as therapeutic for treating kidney stone. This review aims to provide an integrative description on how pentacyclic triterpenes can effectively treat calcium oxalate urolithiasis through various mechanisms such as antioxidant, anti-inflammatory, diuretic, and angiotensin-converting enzyme inhibition. Some of the pentacylic triterpenes which shows promising activities include lupeol, oleanolic acid, betulin, and taraxasterol. Moreover, future perspectives in the development of pentacyclic triterpenes in formulations/drugs for urinary stone prevention are highlighted. It is anticipated that compiled information would serve as a scientific baseline to advocate further investigations on the potential of pentacyclic triterpenes in urolithiasis remediation.

In vitro antilithiatic activity of the hydro-alcoholic extract of Cinnamomum zeylanicum Blume bark on calcium oxalate crystallization.

OBJECTIVE: The study examines the effect of the hydro-alcoholic extract of Cinnamomum zeylanicum Blume bark on crystallization of calcium oxalate. METHODS: The antilithiatic effect of various concentrations of the hydro-alcoholic extract of C. zeylanicum was investigated at various stages of stone formation, using Cystone as a standard reference drug. The effect on calcium oxalate crystallization was evaluated by measuring the change in turbidity over time, during crystal nucleation, growth and aggregation, in the metastable solution of calcium chloride and sodium oxalate. The slope from the change in turbidity over time was measured using a spectrophotometer at 620 and 214 nm in respective tests. The inhibition rate was estimated by comparing turbidity in the presence and absence of extract. Crystals formed under experimental conditions were observed under a light microscope, and number and shape of the crystals were assessed in a randomly selected field. Phytochemical analysis and high-performance thin-layer chromatography of the extract was also carried out. RESULTS: C. zeylanicum significantly reduced crystal nucleation at concentrations of 4, 8 and 10 mg/mL (P < 0.001). The inhibition percentage of crystal growth was between 28.30% and 92.46% in the presence of C. zeylanicum extract and from 20.76% to 64.15% with various concentrations of Cystone. The maximum inhibition of crystal growth was obtained from C. zeylanicum at 2 mg/mL (92.46%). Microscopic examination revealed a reduction in the number and size of crystals. In the aggregation assay, the inhibition percentage of C. zeylanicum was between 16.27% and 100%, while Cystone was from -214.68% to 100% at different concentrations. The highest (100%) inhibition of aggregation was found at 4 mg/mL of both the test and standard drugs. CONCLUSION: We found that C. zeylanicum hydro-alcoholic extract has notable inhibitory effects on various stages of crystallization, in terms of turbidity of solution, as well as the crystal size, number and morphology.

Protective roles of flavonoids and flavonoid-rich plant extracts against urolithiasis: A review.

In the urinary system, urolithiasis is the third prevalent disorder which causes severe pain in individuals. Urinary stones are composed of calcium oxalate (CaOx) and calcium phosphate in approximately 80% of patients. Although various drugs and surgery operations are used to treat the disease, side effects of drugs and the high recurrence after therapy in patients cannot be ignored. Flavonoids are a large group of plant polyphenols with presumed beneficial effects on several common diseases. Whereas, a very few have reached clinical use. The results of recent studies have shown that the plant flavonoids could effectively inhibit the formation of CaOx stones in vitro and in vivo, correlating with their diuretic, antioxidant, anti-inflammatory, antibacterial properties and other protective effects. Thus, the flavonoids or flavonoid-rich plant extracts endowed with anti-urolithiasis activities and probable mechanisms of actions were reviewed. In addition, we also put forward some issues needed to be concerned in future investigations as well as offered prospects and challenges for developing the plant flavonoids into drugs for stone prevention.

In vitro antilithiatic activity of the hydro-alcoholic extract of Cinnamomum zeylanicum Blume bark on calcium oxalate crystallization / 中西医结合学报

OBJECTIVE@#The study examines the effect of the hydro-alcoholic extract of Cinnamomum zeylanicum Blume bark on crystallization of calcium oxalate.@*METHODS@#The antilithiatic effect of various concentrations of the hydro-alcoholic extract of C. zeylanicum was investigated at various stages of stone formation, using Cystone as a standard reference drug. The effect on calcium oxalate crystallization was evaluated by measuring the change in turbidity over time, during crystal nucleation, growth and aggregation, in the metastable solution of calcium chloride and sodium oxalate. The slope from the change in turbidity over time was measured using a spectrophotometer at 620 and 214 nm in respective tests. The inhibition rate was estimated by comparing turbidity in the presence and absence of extract. Crystals formed under experimental conditions were observed under a light microscope, and number and shape of the crystals were assessed in a randomly selected field. Phytochemical analysis and high-performance thin-layer chromatography of the extract was also carried out.@*RESULTS@#C. zeylanicum significantly reduced crystal nucleation at concentrations of 4, 8 and 10 mg/mL (P < 0.001). The inhibition percentage of crystal growth was between 28.30% and 92.46% in the presence of C. zeylanicum extract and from 20.76% to 64.15% with various concentrations of Cystone. The maximum inhibition of crystal growth was obtained from C. zeylanicum at 2 mg/mL (92.46%). Microscopic examination revealed a reduction in the number and size of crystals. In the aggregation assay, the inhibition percentage of C. zeylanicum was between 16.27% and 100%, while Cystone was from -214.68% to 100% at different concentrations. The highest (100%) inhibition of aggregation was found at 4 mg/mL of both the test and standard drugs.@*CONCLUSION@#We found that C. zeylanicum hydro-alcoholic extract has notable inhibitory effects on various stages of crystallization, in terms of turbidity of solution, as well as the crystal size, number and morphology.

The mechanistic insight of polyphenols in calcium oxalate urolithiasis mitigation.

About 12% of world population is affected by different forms of urolithiasis of which the recurrence rate in female is 47-60% and in male is 70-80%. Standard therapeutic agents (allopurinol, citrate, cystone and thiazide diuretics) are used to prevent and treat urolithiasis but these are not universally-effective due to common kidney stone relapse and other side effects. Surgical treatment causes long-term renal damage, hypertension and stone recurrence. Polyphenols, the plant-derived bioactive molecules, have showed protection against cancers, cardiovascular diseases, diabetes, osteoporosis and neurodegenerative diseases, among a number of other ailments. The role of these phytochemicals in urolithiasis management is emerging. Hence, the present review discusses peer-reviewed published literature till date on this aspect and highlights that polyphenols could effectively inhibit the formation of calcium oxalate urinary stones (most common renal stone), correlating with their antioxidant, anti-inflammatory, diuretic and angiotensin-converting enzyme (ACE) inhibition. Further, we have proposed the prospects and challenges in developing the plant polyphenols into drugs against kidney stone prevention. This review might be a stepping stone for further investigation into the clinical implications of the polyphenols in urolithiasis remediation.

In Vitro Antilithiatic Potential of Kalanchoe pinnata, Emblica officinalis, Bambusa nutans, and Cynodon dactylon.

OBJECTIVE: The study aims at the exploration of calcium oxalate (CaOx) crystal growth inhibition potential of Cynodon dactylon, Emblica officinalis, Kalanchoe pinnata, and Bambusa nutans ethyl acetate fraction rich in polyphenol and flavonoid. MATERIALS AND METHODS: Ethyl acetate fraction was separated from the hydromethanolic extract of C. dactylon, E. officinalis, K. pinnata, and B. nutans followed by quantitative analysis for total polyphenol and flavonoid content. Ethyl acetate fraction of all the plants were subjected to in vitro screening for the inhibition of CaO x crystals growth induced by sodium oxalate. RESULTS: The results signify rich presence of polyphenols and flavonoids in K. pinnata and E. officinalis ethyl acetate fractions followed by C. dactylon and B. nutans. Ethyl acetate fractions of B. nutans shoot, E. officinalis fruit, and K. pinnata leaf have excellent in vitro CaO x crystal growth inhibition potential based on both the comparative concentration and the time level to achieve IC50. CONCLUSION: The study outcome substantiates potential in vitro CaO x crystal dissolution and crystal growth inhibition properties of E. officinalis, B. nutans, C. dactylon, and K. pinnata. Rich presence of caffeic acid, ferulic acid, and luteolin in ethyl acetate fraction of B. nutans leaf, and chebulinic acid, chebulagic acid, gallic acid, ellagic acid, and quercetin of E. officinalis may have produced prominent crystal aggregation inhibition response.

The most potent antilithiatic agent ameliorating renal dysfunction and oxidative stress from Bergenia ligulata rhizome.

ETHNOPHARMACOLOGICAL RELEVANCE: The rhizome of Bergenia ligulata is referred by the Ayurvedic system for the treatment of kidney stone since decades and a few, in vitro and in vivo studies also support it. To identify the main phytochemical constituent(s) responsible for antilithiatic activity of its rhizome. MATERIALS AND METHODS: In order to identify the most potent antilithiatic metabolite, the crude extract of rhizome was fractionated using in vitro Calcium oxalate (CaOx) crystal growth inhibitory activity guided fractionation followed by its characterization via LC-MS, FTIR and NMR. Further, the antioxidant potential of purified molecule was assessed using in vitro assays (FRAP and H2O2 scavenging). In vivo activity of the metabolite was evaluated in hyperoxaluric rats given 0.4% ethylene glycol (EG) and 1.0% ammonium chloride (NH4Cl) for 9 days. RESULTS: Activity guided fractionation led to the isolation of most potent antilithiatic metabolite from the rhizome of Bergenia ligulata and spectroscopic analysis revealed it as bergenin. Bergenin showed reducing ability and H2O2 scavenging activity comparable with commercially available anitioxidant, α-tocopherol. At a dose of 10mg/kg body weight of the treated rat, it protected against deleterious effects of lithogenic treatment including weight loss, impaired renal function and oxidative stress, manifested as increased malondialdehyde, reduced redox ratio and decreased antioxidant enzyme activities in the kidneys of hyperoxaluric rats. The creatinine clearance and kidney damage were more improved by bergenin as compared to crude extract of rhizome. CONCLUSIONS: Since, bergenin maintained oxidant/antioxidant balance in hyperoxaluric rats, thus mechanistic insight of its antilithiatic activity was attributed to the antioxidant capability of bergenin. The results of the present study provide significant evidence that bergenin is an active component present in the rhizome of Bergenia ligulata for managing CaOx calculi.