Antiurolithiatic Activity of Ethanolic Extract of Piper cubeba Dried Fruits: An in-vitro and in-vivo Study

Introduction: Piper cubeba is a well-known traditional plant used in unani medicine belonging to the Piperaceae family and has been examined for the treatment of urolithiasis produced by calcium oxalate. Methods: Ethanolic extract of Piper cubeba (EEPC) dried fruits was subjected to phytochemical analysis and HPTLC fingerprinting. An in vitro antiurolithiatic analysis took place through conductometric titrations of CaCl₂ with Na₂C₂O₄. Acute toxicity studies conducted as per OECD guidelines. Urolithiasis was established in rats by supplementing 28 days with 0.75% ethylene glycol in the ingesting water. Beside ethylene glycol, EEPC (100, 200 and 400 mg/kg) was given orally from 15 - 28 days, serum and urine were collected from individual animals and biochemical parameters like BUN, creatinine along with uric acid in serum as well as calcium, oxalate and phosphate in urine the kidney homogenate have been measured on 28^th day. Kidney sections have been organized and histopathologically tested for calcium oxalate crystals. Results: Phytochemical analysis of EEPC disclose the presence of phenolics, tannins, steroids, terpenoids and flavonoids and HPTLC fingerprinting shows the presence of 7 terpenoids, 2 flavonoids when scanned at 540nm and 366nm. In vitro studies showed reduction in CaOx crystal aggregation and promoted nucleation after treatment with EEPC. In vivo studies also showed reduction in elevated levels of serum creatinine, BUN, uric acid, and levels of calcium, oxalate and phosphate in urine and kidney homogenate as compared to disease control rats. The results were supported by histopathological studies. Conclusion: The EEPC have shown significant antiurolithiatic activity by reducing calculi.