"Evaluation of the Litholytic and Nephroprotective Effects of Bryophyllum pinnatum Leaf Extract in an In Vivo Model of Calcium Oxalate Nephrolithiasis" (KEY IJP************557)

• R. Sagarika, Sk. Gousiachandini, K. Joharika, A. Ramya,Falak Naaz,Dr. Jaganath Patro

Abstract

ABSTRACTBackground: Renal calculi, or kidney stones, affect a significant portion of the global population, leading to severe pain, hematuria, and renal dysfunction. Current treatment options include pharmacotherapy, lithotripsy, and surgery, all of which can be costly and recurrent. Herbal alternatives with litholytic properties have gained attention as complementary therapies.Objective: To evaluate the efficacy of Bryophyllum pinnatum leaf extract in the in vivo dissolution and prevention of renal calculi formation.Methods: A controlled in vivo study was conducted using experimental animal models induced with ethylene glycol to promote calcium oxalate stone formation. The animals were treated with aqueous extract of Bryophyllum pinnatum leaves. Biochemical, histological, and urinary analyses were performed to assess antiurolithiatic activity.Results: Administration of Bryophyllum pinnatum showed a significant reduction in urinary oxalate and calcium levels, along with histopathological improvement in renal architecture compared to control.Conclusion: Bryophyllum pinnatum leaf extract demonstrates significant litholytic and nephroprotective activity, supporting its traditional use and potential as a therapeutic candidate for renal calculi.Key wordsBryophyllum pinnatum, renal calculi, kidney stones, anti urolithiatic activity, litholytic effectINTRODUCTIONRenal calculi (kidney stones) remain one of the most prevalent and painful urological disorders worldwide. Epidemiological studies suggest that kidney stones affect approximately 1015% of the global population, with high recurrence rates despite modern interventions 1 2. The most common types of stones, primarily composed of calcium oxalate, are difficult to manage due to their multifactorial aetiology involving diet, dehydration, metabolic disorders, and genetic predisposition 3.Pharmacological treatments often include thiazide diuretics, citrate supplements, or potassium alkali therapy 4. However, these interventions carry potential side effects and may not prevent recurrence, prompting the exploration of alternative natural remedies 5. Traditional medicinal systems such as Ayurveda and Siddha have long employed various herbal remedies for the treatment of urolithiasis6.Bryophyllum pinnatum, also known as Patharchatta in Ayurveda, belongs to the Crassulaceae family and is traditionally used for wound healing, anti-inflammatory, and antiurolithiatic purposes7. Phytochemical investigations have revealed that its leaves are rich in flavonoids, saponins, glycosides, and phenolic compoundsagents known to inhibit crystal aggregation, nucleation, and retention in the renal system 8 9.In recent years, preclinical studies have demonstrated that Bryophyllum pinnatum possesses potential antiurolithiatic activity, particularly in animal models of nephrolithiasis 10 11. These effects are primarily attributed to its diuretic action, antioxidant properties, and its ability to modulate urinary constituents responsible for crystal formation 12.Given the limitations of conventional therapies and the promising pharmacological profile of Bryophyllum pinnatum, this study aims to explore its in vivo efficacy in preventing and treating renal calculi through a scientifically controlled experimental model.MATERIALS AND METHODSStudy Design and Ethical ApprovalThis experimental, controlled in vivo study was conducted in the Department of Pharmacology, Browns College of Pharmacy, following approval from the Institutional Animal Ethics Committee (IAEC), constituted under the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India. All procedures complied with CPCSEA norms for the ethical treatment of laboratory animals.AnimalsA total of 24 healthy adult male Wistar albino rats weighing between 180220 grams were selected for the study. The animals were housed in polypropylene cages under standard laboratory conditions with a 12-hour lightdark cycle, temperature of 222C, and relative humidity of 55%65%. They were fed with a standard pellet diet and had access to water ad libitum.Plant Material and ExtractionFresh leaves of Bryophyllum pinnatum were collected from the local botanical garden and authenticated by a taxonomist from the Department of Botany, insert universityinstitution name. The leaves were washed, shade dried, and coarsely powdered. Aqueous extract was prepared using cold maceration for 48 hours, followed by filtration and concentration using a rotary evaporator at 40C. The extract was stored at 4C until further use.Induction of Renal CalculiUrolithiasis was experimentally induced using a 0.75% vv ethylene glycol (EG) solution administered in drinking water for 28 days, a well-established model for calcium oxalate crystal formation This method promotes hyperoxaluria and mimics human pathological conditions of renal calculi.Experimental GroupingAnimals were randomly divided into four groups (n 6 per group):Group I (Normal Control): Received standard diet and drinking water.Group II (Negative Control): Received 0.75% ethylene glycol in drinking water for 28 days to induce nephrolithiasis.Group III (Standard Group): Received ethylene glycol and Cystone (750mgkg body weight, p.o.) from day 15 to day 28.Group IV (Test Group): Received ethylene glycol and Bryophyllum pinnatum aqueous extract (500mgkg body weight, p.o.) from day 15 to day 28.Biochemical AnalysisAt the end of the treatment period, 24-hour urine samples were collected using metabolic cages. Urine was analyzed for calcium, oxalate, phosphate, magnesium, and creatinine using standard colorimetric methods. Blood was collected via retro-orbital plexus under anaesthesia, and serum was separated for biochemical parameters like blood urea nitrogen (BUN), serum creatinine, and uric acid.Kidney HistopathologyAfter sacrifice, both kidneys were harvested, weighed, and examined grossly. One kidney was fixed in 10% neutral buffered formalin, processed, and embedded in paraffin. Sections of 5m thickness were stained with hematoxylin and eosin (H&E) and evaluated under a microscope for calcium oxalate crystal deposition, tubular necrosis, and inflammatory changes.Statistical AnalysisData were expressed as mean standard deviation (SD). Inter-group comparisons were performed using one-way ANOVA followed by Tukeys post hoc test. A p value of <0.05 was considered statistically significant. Statistical analysis was performed using GraphPad Prism (version 9.0).