Dose-dependant preventive effect of a herbal compound on crystal formation in rat model.

INTRODUCTION: To analyze the dose-dependent preventive effect of a plant-based herbal product on the new crystal formation in a rat model. MATERIALS AND METHODS: A total of 42 rats were divided into 7 groups and zinc discs were placed into the bladder of rats to provide a nidus for the development of new crystal formation: Group 1: control, Group 2: 0.75 percent ethylene glycol (EG); Group 3: 0.75 percent EG plus 0.051 ml of the compound; Group 4: 0.75 percent EG plus 0.179 ml of the compound; Group 5: 0.75 percent EG plus 0.217 ml of the compound; Group 6: 0.75 percent EG plus 0.255 ml of the compound; Group 7 0.75 percent EG plus 0.332 of the compound). The analysis and comparison focused on the disc weights, changes in urinary oxalate and calcium levels, urinary pH, and the histopathologic evaluation of the inflammatory changes in the bladder after 14 days. RESULTS: According to the evaluation of discs placed in the bladders of the animals, animals receiving the herbal compound on a dose-dependent basis showed a limited increase in the disc weights values after 14 days, despite a considerable increase in animals receiving EG alone (p = 0.001). Further evaluation of the increase in disc weights on a dose-dependent basis in different subgroups (from Groups 3 to 7) demonstrated that the limitation of crystal deposition began to be more prominent as the dose of herbal compound increased. This effect was more evident particularly in comparisons between group 7 and others, according to LSD multiple comparison tests (p = 0.001). As anticipated, there has been no discernible change in the weight of the discs in the control group. Although urinary calcium levels in animals of Groups 2, 6, and 7 were significantly higher than the other groups, we were not able to demonstrate a close correlation between urinary oxalate levels and the increasing dose levels. Even though mean urine pH levels were statistically considerably higher in Group 3, there was no statistically significant correlation between the oxalate and calcium levels between all groups, and no association was seen with the administration of herbal agents. The transitional epithelium between the three groups of animals' bladder samples did not exhibit any appreciable difference according to pathological analysis. CONCLUSIONS: In this animal model, the treatment of the compound was successful in lowering the amount of crystal deposition surrounding the zinc discs, most noticeably at a dosage of 0.332 ml, three times per day.

Effects of high-dose uric acid on cellular proteome, intracellular ATP, tissue repairing capability and calcium oxalate crystal-binding capability of renal tubular cells: Implications to hyperuricosuria-induced kidney stone disease.

Hyperuricosuria is associated with kidney stone disease, especially uric acid (UA) and calcium oxalate (CaOx) types. Nevertheless, detailed mechanisms of hyperuricosuria-induced kidney stone formation remained unclear. This study examined changes in cellular proteome and function of renal tubular cells after treatment with high-dose UA for 48-h. Quantitative proteomics using 2-DE followed by nanoLC-ESI-ETD MS/MS tandem mass spectrometry revealed significant changes in levels of 22 proteins in the UA-treated cells. These proteomic data could be confirmed by Western blotting. Functional assays revealed an increase in intracellular ATP level and enhancement of tissue repairing capability in the UA-treated cells. Interestingly, levels of HSP70 and HSP90 (the known receptors for CaOx crystals) were increased in apical membranes of the UA-treated cells. CaOx crystal-cell adhesion assay revealed significant increase in CaOx-binding capability of the UA-treated cells, whereas neutralization of the surface HSP70 and/or HSP90 using their specific monoclonal antibodies caused significant reduction in such binding capability. These findings highlighted changes in renal tubular cells in response to high-dose UA that may, at least in part, explain the pathogenic mechanisms of hyperuricosuria-induced mixed kidney stone disease.

Preparation and characterization of selenized Astragalus polysaccharide and its inhibitory effect on kidney stones.

Astragalus polysaccharide (APS) was modified using the Na2SeO3/HNO3 method to obtain selenized APS (Se-APS) with a selenium content of 1.75 mg/g. The structure and physicochemical properties of APS and Se-APS were investigated through transmission electron microscopy-energy dispersive spectroscopy mapping, fourier transform infrared spectroscopy, nuclear magnetic resonance, nano-zetasizer analysis, atomic force microscopy, and scanning electron microscopy. APS and Se-APS did not exhibit toxic effects on human kidney proximal tubular epithelial (HK-2) cells and were able to remove hydroxyl and DPPH radicals, alleviate the damage caused by calcium oxalate (CaOx) monohydrate (COM) crystals to HK-2 cells, reduce intracellular reactive oxygen species levels, and restore cell viability and morphology. Both APS and Se-APS could inhibit COM growth, induce calcium oxalate dihydrate formation, and increase the absolute zeta potential of the crystals to inhibit crystal aggregation. However, the ability of Se-APS to regulate CaOx crystals and protect the cells from COM-induced damage was better than that of APS. These results suggested that Se-APS might be a candidate drug for the treatment and prevention of kidney stones.

A Physiologically Based in Silico Tool to Assess the Risk of Drug-Related Crystalluria.

Drug precipitation in the nephrons of the kidney can cause drug-induced crystal nephropathy (DICN). To aid mitigation of this risk in early drug discovery, we developed a physiologically based in silico model to predict DICN in rats, dogs, and humans. At a minimum, the likelihood of DICN is determined by the level of systemic exposure to the molecule, the molecule's physicochemical properties and the unique physiology of the kidney. Accordingly, the proposed model accounts for these properties in order to predict drug exposure relative to solubility along the nephron. Key physiological parameters of the kidney were codified in a manner consistent with previous reports. Quantitative structure-activity relationship models and in vitro assays were used to estimate drug-specific physicochemical inputs to the model. The proposed model was calibrated against urinary excretion data for 42 drugs, and the utility for DICN prediction is demonstrated through application to 20 additional drugs.

Delving into the Antiurolithiatic Potential of Tribulus terrestris Extract Through -In Vivo Efficacy and Preclinical Safety Investigations in Wistar Rats.

Modern treatment interventions for kidney stones are wrought with side-effects, hence the need for alternative therapies such as plant-based medicines. We have previously documented through in vitro studies that statistically optimized aqueous extract of Tribulus terrestris (Zygophyllaceae family) possesses antiurolithic and antioxidant potential. This provides strong scientific foundation to conduct in vivo efficacy and preclinical safety studies to corroborate and lend further proof to its ability to prevent and cure kidney stones. The preventive and curative urolithiatic efficacy in experimentally induced nephrolithiatic Wistar rats, along with preclinical toxicity was evaluated following oral administration of statistically optimized aqueous extract of T. terrestris. Treatment showed augmented renal function, restoration of normal renal architecture and increase in body weight. Microscopic analysis of urine revealed excretion of small sized urinary crystals, demonstrating that treatment potentially modulated the morphology of renal stones. Tissue enzymatic estimation affirmed the antioxidant efficacy of treatment with reduced free radical generation. Significant upregulation of p38MAPK at both the gene and protein level was noted in hyperoxaluric group and interestingly treatment reversed it. Acute oral toxicity study established the Median Lethal Dose (LD50) to be greater than 2000 mg/kg body weight (b.wt.) No observed adverse effect level (NOAEL) by repeated oral toxicity for 28 days at 750 mg/kg b.wt. was noted. This study lends scientific evidence to the safe, preventive and curative potential of statistically optimized aqueous extract of T. terrestris at a dose of 750 mg/kg b.wt. and suggests that the extract shows promise as a therapeutic antiurolithic agent.

Vitamin D and Calcium Supplementation Accelerates Randall's Plaque Formation in a Murine Model.

Most kidney stones are made of calcium oxalate crystals. Randall's plaque, an apatite deposit at the tip of the renal papilla, is considered to at the origin of these stones. Hypercalciuria may promote Randall's plaque formation and growth. We analyzed whether long-term exposure of Abcc6-/- mice (a murine model of Randall's plaque) to vitamin D supplementation, with or without a calcium-rich diet, would accelerate the formation of Randall's plaque. Eight groups of mice (including Abcc6-/- and wild type) received vitamin D alone (100,000 UI/kg every 2 weeks), a calcium-enriched diet alone (calcium gluconate 2 g/L in drinking water), both vitamin D supplementation and a calcium-rich diet, or a standard diet (controls) for 6 months. Kidney calcifications were assessed by 3-dimensional microcomputed tomography, µ-Fourier transform infrared spectroscopy, field emission-scanning electron microscopy, transmission electron microscopy, and Yasue staining. At 6 months, Abcc6-/- mice exposed to vitamin D and calcium supplementation developed massive Randall's plaque when compared with control Abcc6-/- mice (P < 0.01). Wild-type animals did not develop significant calcifications when exposed to vitamin D. Combined administration of vitamin D and calcium significantly accelerates Randall's plaque formation in a murine model. This original model raises concerns about the cumulative risk of vitamin D supplementation and calcium intakes in Randall's plaque formation.

Local Anesthetic Flexible Ureterorenoscopy in the Management of Urolithiasis.

Introduction: Patients unfit for general anesthesia who present with renal tract pathology currently have limited options. Many of these patients present in the emergency setting with imperative reasons for intervention, including sepsis, renal failure, and pain. Conservative management and temporizing measures, such as percutaneous nephrostomy, are associated with significant morbidity. Ureterorenoscopy (URS) is a central component of the management of upper tract disease and is routinely performed under general anesthesia. We describe our institution's experience of URS using only local anesthetic (LA) lubricating gel per urethra. Methods: A single centre, retrospective analysis of 78 patients was performed for an 11 year period. Demographic data and Charlson comorbidity index scoring were collected for all patients. Outcomes, including stone-free rates, tolerability, and complications, were analyzed. Results: In total 58% of patients were men. Mean age was 68 and Charlson comorbidity index was 5.2. Indications for URS included pain (68%) and renal failure (15%). Totally 10% of patients previously had retrograde stenting because of sepsis. Median stone size was 8 mm. All patients were able to tolerate the procedure and none were abandoned because of pain. The overall stone-free rate was 82% after one procedure. The stone-free rate for mid and distal ureteral stones was 97%. Nineteen percent of patients were left with a ureteral stent after the procedure, with the remaining patients left totally tubeless. Median length of stay was 1 day. There were no complications above Clavien Grade 2. Conclusion: Urologists are increasingly faced with unfit patients presenting with urolithiasis. In the appropriately selected patient, LA flexible ureterorenoscopy is a feasible option with good outcomes. This approach is a useful addition to the armamentarium available to patients deemed unsuitable for general or regional anesthesia.

Metabolomic analysis reveals a protective effect of Fu-Fang-Jin-Qian-Chao herbal granules on oxalate-induced kidney injury.

Nephrolithiasis is one of the world's major public health burdens with a high incidence and a risk of persistent renal dysfunction. Fu-Fang-Jin-Qian-Chao granules (FFJQC), a traditional Chinese herb formula, is commonly used in treatment of nephrolithiasis. However, the therapeutic mechanism of FFJQC on kidney stone has still been a mystery. The objective of the present study is to explore the therapeutic mechanism of FFJQC on kidney injury and identify unique metabolomics patterns using a mouse model of kidney stone induced by a calcium oxalate (CaOx) deposition. Von Kossa staining and immuno-histopathological staining of osteopontin (OPN), cluster of differentiation 44 (CD44) and calbindin-D28k were conducted on renal sections. Biochemical analysis was performed on serum, urine, and kidney tissues. A metabolomics approach based on ultra-HPLC coupled with quadrupole-TOF-MS (UHPLC-Q-TOF/MS) was used for serum metabolic profiling. The immunohistopathological and biochemical analysis showed the therapeutic benefits of FFJQC. The expression levels of OPN and CD44 were decreased while calbindin-D28k increased after the CaOx injured mice were treated with FFJQC. In addition, total of 81 serum metabolites were identified to be associated with protective effects of FFJQC on CaOx crystal injured mice. Most of these metabolites were involved in purine, amino acid, membrane lipid and energy metabolism. Potential metabolite biomarkers were found for CaOx crystal-induced renal damage. Potential metabolite biomarkers of CaOx crystal-induced renal damage were found. FFJQC shows therapeutic benefits on CaOx crystal injured mice via regulation of multiple metabolic pathways including amino acids, purine, pyrimidine, glycerolipid, arachidonic acid (AA), sphingolipid, glycerophospholipid, and fatty acid.

Urothelium proliferation is a trigger for renal crystal deposits in a murine lithogenesis model.

Most mouse kidney stone models induce nephrocalcinosis rather than urolithiasis. The aim of our study was to find an accelerated experimental model in order to study the early events of stone formation, that is, at the time of crystal binding to intrarenal urothelium. C57B6 mice exposed to vitamin D supplements and water containing hydroxyl-L-proline, ammonium chloride and calcium chloride were studied for 42 days. A group receiving urothelial cell mitogen Fibroblast Growth Factor 7 (FGF7) was compared to control group receiving saline. Calcium oxalate monohydrate (COM) crystals were detected in urines by day 2 and within urinary spaces in specialized fornix areas in both groups as soon as day 14 with enhanced deposits in FGF7 group compared to controls at day 21. Urothelial cells proliferation, uroplakin III downregulation and de novo expression of osteopontin receptor CD44 detected in FGF7 group, were delayed in the control group (day 42). Crystal aggregates within specialized fornix areas by day 42 were located in urinary spaces but also within and under a multilayered metaplastic urothelium, simultaneous to macrophages influx. Point of note, administration of a normal diet by day 21 was responsible for a spontaneous crystal clearance. Our data show that under supersaturation conditions, urothelial cell proliferation and calcium oxalate crystal retention occur within specialized fornix areas. Enhanced crystal deposits following FGF7 administration suggest that urothelium proliferation would be a relevant trigger for renal stone formation.

Effect of Polygonum Aviculare L. on Nephrolithiasis Induced by Ethylene Glycol and Ammonium Chloride in Rats.

PURPOSE: Nephrolithiasis is a common urinary tract disease, in addition to the pain and treatment costs, there may be significant complications resulting from the stones. This study intended to investigate the effects of Polygonum Aviculare L. aqueous extract (PAE) on urolithiasis induced by ethylene glycol (EG) and ammonium chloride (AC) in rats. MATERIALS AND METHODS: Sixty-four male Wistar rats were randomly divided into eight groups (n = 8). Rats in the normal control group (I) received no treatment. The sham groups (III and IV) were given PAE. at 100 and400 mg/kg by gavage for 28 days. The disease control group (II), the prevention groups ( V and VI), and the therapeutic groups (VII and VIII), received 1% EG and .25 AC in their drinking water for 28 days. The prevention groups (from the start of EG administration), and the therapeutic groups (from the 14th day of EG administration),received PAE at 100 and 400 mg/kg by gavage. At the end of the experiment, kidneys were examined for CaOx deposits and tubulointerstitial changes. RESULTS: The number of CaOx crystals and tubulointerstitial changes increased significantly in group II rats compared to groups I, III, and IV (P < .001). The number of CaOx crystals (P < .001) and tubulointerstitial changes (P < .001) in the prevention groups, and the number of CaOx crystals (P < .05) and interstitial changes (P < .05) inthe therapeutic groups declined significantly compared to group II. CONCLUSION: Results show aqueous extract of Polygonum Aviculare L. is effective in the prevention and treatment of kidney stones.

Therapeutic Effects of Aqueous Extracts of Cerasus Avium Stem on Ethylene Glycol- Induced Kidney Calculi in Rats.

PURPOSE: To investigate the therapeutic effects of the aqueous extract of Cerasus Avium stem on kidney calculi. MATERIALS AND METHODS: In this experimental study, forty-eight (48) male Wistar rats were randomly allocated into six (6) groups and were studied during a 30 day period. Group A served as normal control and Group B received 1% ethylene glycol in drinking water (EG group). C, D, E, and F Groups, received 1% ethylene glycol from day 1 and were used as prevention and treatment subjects. Rats in prevention groups of low dose (C) and high dose (D) extract, were gavaged with 200 and 400 mg/kg extract respectively from first day of the experiment and treatmentgroups of low dose (E) and high dose (F) extract, were gavaged with 200 and 400 mg/kg extract respectively from the 15th day of the experiment. RESULTS: On the 30th days of the experiment, serum level of magnesium and potassium decreased significantly in EG group compared with A,C,D,E and F groups (P < .05), while serum level of calcium, creatinine, uric acid, sodium and urine level of calcium, creatinine, uric acid, increased significantly in EG group compared with A,C,D,E and F groups (P < .05). In the prevention and treatment groups, the number of deposits decreased significantly compared with EG group on the 30th day (P < .05). CONCLUSION: Cerasus Avium stem has a therapeutic effect on calcium oxalate stones in rats with nephrolithiasis and reduces the number of calcium oxalate deposits.

Antiurolithiasis Activity of Bioactivity Guided Fraction of Bergenia ligulata against Ethylene Glycol Induced Renal Calculi in Rat.

Dried rhizome of Bergenia ligulata (pashanbhed) is commonly used as a traditional herbal medicine with a wide range of therapeutic applications including urolithiasis. Aqueous extract of B. ligulata was prepared through maceration followed by decoction (mother extract, 35.9% w/w). Further, polarity based fractions were prepared successively from mother extract which yielded 3.4, 2.9, 5.4, 7.5, and 11.3% w/w of hexane, toluene, dichloromethane (DCM), n-butanol, and water fractions, respectively. The in vitro, ex vivo, and real-time antiurolithiasis activity of mother extract and fractions were carried out using aggregation assay in synthetic urine and in rat plasma. The study revealed that DCM fraction has significantly (p < 0.05) greater inhibitory potential than other fractions. Ethylene glycol in drinking water (0.75%, v/v) for 28 days was used for induction of urolithiasis and the curative effects of mother extract and DCM fraction were checked for the level of oxalate, calcium, creatinine, uric acid, and urea of both urine and serum. Treatment with mother extract and DCM fraction at a dose of 185 mg/kg and 7 mg/kg, respectively, in ethylene glycol induced rats resulted in a significant (p < 0.05) decrease in serum and urine markers. Histological study revealed lower number of calcium oxalate deposits with minimum damage in the kidneys of mother extract and DCM fraction treated rats. This result provides a scientific basis for its traditional claims.

Detection of carcinogenic metals in kidney stones using ultraviolet laser-induced breakdown spectroscopy.

The UV single-pulsed (SP) laser-induced breakdown spectroscopy (LIBS) system was developed to detect the carcinogenic metals in human kidney stones extracted through the surgical operation. A neodymium yttrium aluminium garnet laser operating at 266 nm wavelength and 20 Hz repetition rate along with a spectrometer interfaced with an intensified CCD (ICCD) was applied for spectral analysis of kidney stones. The ICCD camera shutter was synchronized with the laser-trigger pulse and the effect of laser energy and delay time on LIBS signal intensity was investigated. The experimental parameters were optimized to obtain the LIBS plasma in local thermodynamic equilibrium. Laser energy was varied from 25 to 50 mJ in order to enhance the LIBS signal intensity and attain the best signal to noise ratio. The parametric dependence studies were important to improve the limit of detection of trace amounts of toxic elements present inside stones. The carcinogenic metals detected in kidney stones were chromium, cadmium, lead, zinc, phosphate, and vanadium. The results achieved from LIBS system were also compared with the inductively coupled plasma-mass spectrometry analysis and the concentration detected with both techniques was in very good agreement. The plasma parameters (electron temperature and density) for SP-LIBS system were also studied and their dependence on incident laser energy and delay time was investigated as well.

Chronic trimethyltin chloride exposure and the development of kidney stones in rats.

We recently reported that occupational exposure to trimethyltin (TMT) is a risk factor for developing kidney stones. To further examine the association between TMT exposure and the formation of kidney stones, we conducted a 180-day animal study and exposed the randomly grouped Sprague-Dawley (SD) rats to TMT in the drinking water at doses of 0, 8.2, 32.8 and 131.3 µg kg(-1) day(-1). Transient behavioral changes were observed in the high-dose group during the first 2 weeks of exposure. TMT exposure led to a significant dose-dependent inhibition of renal H(+)/K(+)-ATPase and an increase in urinary pH. In comparison to no kidney stones being identified in the control and the lowest dose group, 1 rat in the 32.8 µg kg(-1) day(-1) dose group and 3 out of 9 rats in the 131.3 µg kg(-1) day(-1) dose group were found to have stones in the kidney/urinary tract. Pathological analysis showed that more wide spread calcium disposition was observed in kidneys of rats with TMT exposure compared with the rats in the control group. However, X-ray diffraction (XRD) analysis found that the kidney stones were mainly composed of struvite with the formula: NH4MgPO4 6H2O, while calcium-containing components were also detected. Together, this study further demonstrates through animal studies that chronic exposure to a relatively low level of TMT induces nephrotoxicity and increases the risk for developing kidney stones.

Antioxidant therapy prevents ethylene glycol-induced renal calcium oxalate crystal deposition in Wistar rats.

PURPOSE: Renal epithelial cell injury by reactive oxygen species is a prerequisite step in the pathogenesis of urolithiasis, and there is increasing evidence that reactive oxygen species is produced and oxidative stress (OS) is developed during idiopathic calcium oxalate nephrolithiasis. It appears that the administration of natural antioxidants has been used to protect against nephrolithiasis in human and experimental animals. METHODS: Calcium oxalate urolithiasis was induced experimentally by administration of 0.75 % v/v ethylene glycol in drinking water of male Wistar rats weighing 150-200 g. Study was conducted in 4- and 8-week periods. In the 4-week period, Group 1 (control) was fed a standard commercial diet. Group 2 received the same diet with the addition of 0.75 % of ethylene glycol (EG). Group 3 received EG plus the diet, and water with additional antioxidant nutrients, and lemon juice as the dietary source of citrate (EG + AX). Group 4 was the same as Group 3, but with no EG in water. In the 8-week study protocol, Group 5 was fed the standard diet with EG in water for the first 28 days, followed with no EG. Group 6 (curative group) received the diet with EG for the first 28 days, followed by discontinuation of EG plus the addition of antioxidant nutrients. Group 7 was provided the diet with antioxidant nutrients for 8 weeks. Group 8 (preventive group) received the diet with antioxidant nutrients for 4 weeks, followed by antioxidant nutrients with EG for the next 4 weeks. Lime juice was given along the antioxidants. After treatment period, kidneys were removed and used for histopathological examination. RESULTS: In the 4-week study, the mean number of crystal deposits in Group 2 was significantly higher than that of animals in Group 3. After 8 weeks, animals given curative antioxidant supplementation within the second 4-week period developed fewer deposits in Group 6 as compared to Group 5 animals. In the other preventive AX loading Group 8, the number of crystal deposits was substantially less than that of either Group 2 or Group 5 animals (EG-treated rats). CONCLUSION: Results showed a beneficial effect on treating and superior renal protection for preventing stone deposition in the rat kidney. These results provide a scientific rationale for preventive and treatment roles of antioxidant nutrient complex in human kidney stone disease.

Catechin prevents the calcium oxalate monohydrate induced renal calcium crystallization in NRK-52E cells and the ethylene glycol induced renal stone formation in rat.

BACKGROUND: Reactive oxygen species play important roles in renal calcium crystallization. In this study, we examined the effects of catechin, which have been shown to have antioxidant properties on the renal calcium crystallization. METHODS: In the vitro experiment, the changes of the mitochondrial membrane potential, expression of superoxide dismutase (SOD), 4-hydroxynonenal (4-HNE), cytochrome c, and cleaved caspase 3 were measured to show the effects of catechin treatment on the NRK-52E cells induced by calcium oxalate monohydrate (COM). In the vivo study, Sprague-Dawley rats were administered 1% ethylene glycol (EG) to generate a rat kidney stone model and then treated with catechin (2.5 and 10 mg/kg/day) for 14 days. The urine and serum variables were dected on 7 and 14 days after EG administration. The expression of cytochrome c, cleaved caspase 3, SOD, osteopontin (OPN), malondialdehyde (MDA), 8-hydroxy-2'-deoxyguanosine (8-OHdG) in kidney were measured. Furthermore, the mitochondrial microstructure in the kidney was also examined by transmission electron microscopy. RESULTS: Catechin treatment could prevent the changes in mitochondrial membrane potential and expression of SOD, 4-HNE, cytochrome c, and cleaved caspase 3 in NRK-52E cells induced by the COM. For the in vivo experiments, the EG administration induced renal calcium crystallization was also prevented by the catechin. The expression of SOD, OPN, MDA, OPN and 8-OHdG, were increased after EG administration and this increase was diminished by catechin. Moreover, catechin also prevented EG induced mitochondrial collapse in rat. CONCLUSIONS: Catechin has preventive effects on renal calcium crystallization both in vivo and in vitro, and provide a potential therapeutic treatment for this disease.

Whole Leea macrophylla ethanolic extract normalizes kidney deposits and recovers renal impairments in an ethylene glycol-induced urolithiasis model of rats.

OBJECTIVE: To investigate the antilithiatic effect of the whole Leea macrophylla (L. macrophylla) Roxb (Leeaceae) ethanol extract in ethylene glycol-induced urolithiasis model of rats. METHODS: Forty two seven weeks old male wistar albino rats were randomly divided into two major groups namely: preventive (n=18) and therapeutic (n=24). Preventive group was further subdivided into 3 groups of 6 rats namely: preventive control (PC), preventive lithiatic control (PLC) and preventive lithiatic L. macrophylla (PLLM). Similarly, therapeutic group was also subdivided into 4 groups of 6 rats namely: therapeutic control (TC), therapeutic lithiatic control (TLC), therapeutic lithitatic L. macrophylla (TLLM) and therapeutic lithiatic cystone (TLCYS). The lithiasis was induced by 0.75% (v/v) ethylene glycol in the drinking water of all groups except the PC and TC groups. The urinary ionic parameters such as calcium, inorganic phosphate, oxalate, magnesium & creatinine and renal morphology were altered by ethylene glycol, which were partially recovered by 14 d preventive and almost fully recovered by 28 d therapeutic intervention trials with L. macrophylla extract (500 mg/kg BW daily). RESULTS: Significant difference on recovery was observed between preventive and therapeutic interventional trials. Anti-urolithiatic effect of cystone was significantly (P<0.001) higher than extracts. L. macrophylla extract was found nontoxic in the acute toxicity test. CONCLUSION: The results of this study demonstrated very promising anti-urolithiatic effect of L. macrophylla extract with preventive and therapeutic treatments in this experimental condition.

Anti-urolithiatic effects of Punica granatum in male rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional use of Punica granatum has been reported to regulate urine discharge and controls the burning sensation of urine. MATERIALS AND METHODS: Animals model of calcium oxalate urolithiasis was developed in male rats by adding ethylene glycol 0.75% in drinking water. The Punica granatum chloroform extract (PGCE) and Punica grantum methanol extract (PGME) orally at 100, 200 and 400mg/kg, respectively, were administered along with ethylene glycol for 28 days. On 28 day, 24h urine was collected from individual rats and used for estimation of urine calcium, phosphate and oxalate. The serum creatinine, urea and uric acid levels were estimated in each animal. The kidney homogenate was used for the estimation of renal oxalate contents. The paraffin kidney sections were prepared to observe the CaOx deposits. RESULTS: The ethylene glycol control (Gr.-II) had significant (P<0.001 vs. normal) increase in levels of urine oxalate, calcium and phosphate, serum creatinine, urea and uric acid and renal tissues oxalates, as compared to normal (Gr.-I). The paraffin kidney sections show significant histopathological changes. The treatment of PGCE and PGME at 100, 200 and 400mg/kg doses, significantly (P<0.001 vs. control) decreased the urine oxalate, calcium and phosphate, renal tissue oxalates and serum creatinine, urea and uric acid, in EG induced urolithiasis after 28 days. CONCLUSIONS: The PGCE and PGME at the doses of 400mg/kg, found to be more effective in decreasing the urolithiasis and regeneration of renal tissues in male rats.

[Influence of electroacupuncture combined with intensive moxibustion intervention on pathological changes of kidney and ureter tissues, plasma and uterine Ca2+, creatinine, and urea nitrogen levels in calcium oxalate stone rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation combined with moxibustion (MOX) on changes of the kidney and ureter tissue, plasma and uterine Ca2+, creatinine (Or) and urea nitrogen (UN) concentrations in rats with renal calcium oxalate stone, so as to reveal its mechanism underlying relieving kidney stone. METHODS: A total of 60 Wistar rats were randomized into control group, model group, EA plus MOX of "Pangguangshu" (BL 28), "Sanyinjiao" (SP 6) and "Zhishi" (BL 52) (routine EA-MOX group), EA-intensive (INT) MOX of "Shenshu" (BL 23, EA-INT-MOX-BL 23 group) and EA-INT-MOX-Ashi group (12 rats/group). Renal calcium oxalate stone model was established by feeding the animalwith 1% ethylene alcohol and 1% ammonium chloride for 7 days. EA was applied to BL 28, SP 6, BL 52, BL 23, and Ashi-point for 15 min, respectively. For rats of the EA-INT-MOX-BL 23 group, EA intervention was applied to the superficial, medium and deep layers of BL 23 for 5 min, respectively, and INT MOX was given for 11 - 13 min. Plasma and uterine Ca2+ conctents were detected by absorption spectrometry, plasma and urine Cr contents determined by sarcosine oxidase method. Blood UN (BUN) level was determined by enzyme coupling rate method and uric acid (UA) detected by photoelectric colorimetry method. RESULTS: Compared with the control group, inflammatory cell infiltration, congestion, space enlargement in the glomerulus and renal interstitium, and unsmooth of the ureter wall with exfoliated necrosis tissue in the ureter were seen in the model group. These situations were relatively milder in the routine EA-MOX, EA-INT-MOX-BL 23 and EA-INT-MOX-Ashi groups. In comparison with the control group, plasma and uterine Ca2+ Cr, BUN, and UA levels were increased significantly in the model group (P < 0.01). While compared to the model group, plasma and uterine Ca2+ Cr, BUN, and UA levels were down-regulated considerably (P < 0.05, P < 0.01) in the three treatment groups, and the effects of both EA-INT-MOX-BL 23 and EA-INT-MOX-Ashi groups were significantly superior to those of the routine EA-MOX group (P < 0.05, P < 0.01). CONCLUSION: EA plus intensive moxibustion can effectively lower plasma and uterine Ca2+, Cr, BUN and UA levels, and improve pathological changes of the kidney in kidney stone rats, which may contribute to its effect in bettering renal function. The effect of EA plus intensive moxibustion is markedly better than that of routine EA plus moxibustion in lowering plasma and urine biochemical indexes.