Basil (Ocimum basilicum L.) and/or Celery (Apium graveolens L.) Leaves Aqueous Extracts Role in Opposition to Drinking Contaminated Water Induced Male Rats Urinary Stones and Renal Deteriorations

Abstract

Aims: The current research was designed to explore the role of basil and/or celery leaves aqueous extracts against urinary stones (urolithiasis) and renal deteriorations induced by drinking water contaminated with ethylene glycol (EG) and ammonium chloride (AC) in male rats.
 Place and Duration of Study: The Medical Research Center of the Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt. The experiment duration was 35 days, 7 days for acclimatization followed by 28 days experimentation.
 Methodology: Fifty-five adult male Sprague-Dawley rats were classified randomly into 5 groups of ten animals each, except urolithiatic control group which contained 15 rats. Rats were treated as follow: Group I: healthy control group (HCG), rats drank distilled water and received a placebo 1 ml distilled water daily by intra-gastric tube ; Group II: urolithiatic control group (UCG), rats drank distilled water containing ( 0.75% EG and 1% AC ) and the access to water was ad libitum and received 1 ml distilled water daily by intra-gastric tube.; Group III : urolithiatic rats supplemented with aqueous basil extract(U+ABE), rats drank distilled water containing ( 0.75% EG and 1% AC) and the access to water was ad libitum and supplemented with (200mg/kg body weight) aqueous basil extract daily by intra-gastric tube.; Group IV: urolithiatic rats supplemented with aqueous celery extract(U+ACE), rats drank distilled water containing ( 0.75% EG and 1% AC) and the access to water was ad libitum and supplemented with (200mg/kg body weight) aqueous celery extract daily by intra-gastric tube. ; Group V: urolithiatic rats supplemented with aqueous basil and celery extract(U+ABE+ACE) , rats drank distilled water containing ( 0.75% EG and 1% AC) and the access to water was ad libitum and supplemented with (100mg/kg body weight) aqueous basil extract and (100mg/kg body weight) aqueous celery extract daily by intra-gastric tube. At the end of the study 28 days, 24 h urine samples were collected from each individual animal and then rats were sacrificed under sodium barbiturate anesthesia. Blood samples were collected from the hepatic portal vein, serum was separated for biochemical analyses. Kidney samples were separated for tissue homogenate preparation, urinary bladder and ureters were separated and opened to collect urinary calculi.
 Results: Research results documented that basil and celery aqueous extracts contain significant amount of active constituents including flavonoids and polyphenols. Drinking water contaminated with EG and AC caused significant decrease (p≤0.01) in urine volume, pH, urinary and renal magnesium levels with significant increase (p≤0.01) in urinary and renal calcium, phosphate and oxalate levels. It also caused significant increase in kidney function tests [serum creatinine, uric acid, urea, as well as cystatin C] levels .Also renal enzyme activities of [acidic phosphatase (ACP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP)] activities were significantly increased (p≤0.01) with significant decrease in renal lactate dehydrogenase (LDH) activity. Water contamination also induced renal mitochondrial dysfunction (decreased complex-1 activity), apoptosis (increased caspase-3 activity) in association with a state of renal oxidative stress [increased renal malondialdehyde (MDA) level related to decreased renal reduced glutathione (GSH)level and superoxide dismutase (SOD) activity].Inflammation [increased renal tumor necrosis factor- alpha (TNF- α) and interleukin 18 (IL-18)] with suppressed immune function [decreased serum immunoglobulin (IgG and IgM)] levels were observed in urolithiatic control group. As a result of biochemical changes resulted from EG and AM administration mineral precipitation and urinary stones formation was observed in urolithiatic control group. Oral administration of aqueous basil and /or celery extracts to urolithiatic rats improved urine analysis parameters, renal functions, renal enzymes activities and mitochondrial function. Aqueous extracts also counteracted renal oxidative stress, inflammation, suppressed immune response and urinary stones formation.
 Conclusion: Research results proved that aqueous basil and/or celery extracts counteracted and ameliorated urinary stones formation and renal deteriorations associated with drinking water contaminated with EG and AC due to their content of active constituents including flavonoids and polyphenols. The most significant improvements were recorded in the group supplemented with both extracts.