Influence of Lycopene on Renal Tissue Injury in Celebrex Induced Albino Rats; An Experimental Research Study

Objective: To analyze the renal tissue injury in Celebrex induced albino rats with improvement by lycopene. Study design: Experimental research study. Abode of study: Animal House, Basic medical sciences institute, Karachi, Materials and Methods: Celebrex and antioxidant agent lycopene were used in this experimental study. Drugs were administered orally to forty male albino rats weighing of around 250gm for research. Rats were retained in isolated cages and alienated into 4 sets including control, diseased, treated groups. The drugs were administered by gastric tube once24-hourly for four weeks. After finalization of research work, rats were sacrificed and body tissues were preserved for staining. Results: gB Gomori’s calcium phosphate stained sections showed marked deposition of dark black particles of alkaline phosphate in cellular cytoplasm of proximal convoluted tubules but gC showed scarce deposition of black particles of alkaline phosphate in the cellular cytoplasm of proximal Original Research Article Sundus et al.; JPRI, 34(25A): 14-19, 2022; Article no.JPRI.84423 15 convoluted tubules. Conclusion: This experiment discloses that lycopene recuperate the raised level of alkaline phosphate in the cytoplasm of proximal convoluted tubules.


INTRODUCTION
Non-Steroidal Anti-inflammatory drugs are the number one most frequently recommended medicine & COX enzyme inhibitor for various ailments like inflammation, fever, arthritis and painful illnesses. They further categorize on the basis of (COX) enzyme selectivity. One group inhibits both COX 1 &2 whereas second group is only cox-2 inhibitor [1]. Non-selective group raise the risk of gastric ulcers and intestinal bleeding due to weakening of prostaglandin-dependent mucosal protective ability because NSAIDs inhibits prostaglandins synthesis [2,3]. PGE 2 regulates the fluid metabolism and hemodynamic in renal tissue and its inhibition plays a significant role in inflammation, peptic ulcer ailment, renal dysfunction and impairment of intestinal mucosa [4]. Myeloperoxidase enzyme (MPO) found in neutrophils and macrophages plays an important role in inflammation and oxidative stress [5,6]. Organs in which Xenobiotic metabolism occur are kidney, liver & GIT. It is the type of metabolism in which biochemical alteration of drug take place by enzymes. Celebrex is COX-2 enzyme inhibitor & COX-2 enzyme mainly affect salt and water excretion, so it has toxic effect on kidney.  [15,16]. Lycopene found in blood plasma anti-carcinogenic, antiinflammatory and antioxidant in character. Inhibits prostaglandin synthesis & also prevents various malignancies, Alzheimer's ailment, & cardiovascular illnesses, as well as reduces ageing [17]. It prevents cellular damage due to scavenging properties of reactive oxygen species, singlet oxygen-quenching capability, several conjugated double bonds & reacts with reactive oxygen species to decrease DNA mutilation [18,19,20]. It also prevents lipid peroxidation in cellular basement membrane [21,22,23]. It detoxifies toxins by antioxidation, anti-inflammation activitiy [24,25]. The experimental research study was organized to examine the advantageous effects of lycopene in comparison with injurious effects of Celebrex.

MATERIALS AND METHODS
This is a 4 weeks experimental research study, came about in JPMC (Jinnah postgraduate medical center), Karachi along with endorsement from ethical review board No.F-1-2/BMSI-E.COMT/039/JPMC. 40 fully grown up healthy albino rats, three to four months old, around 250gm were taken from USA lab and fostered in animal house in isolated cages. Assessed for 7 days for their health condition and weighed former the beginning of research and isolated into 4 sets (control, diseased, treated) and dosage calculated according to their weight. Tablet Celebrex purchased from Getz Pharma & lycopene pigment capsule from General Nutrition Corporation [26,27], were administered by gastric tube 0.05g/kg orally once24-hourly for four weeks. After finalization of research work, rats were sacrificed and body tissues were preserved for staining. gA= control. gB=: Celebrex 0.05g/kg per oral. (Morbid group) gC= Celebrex with lycopene pigment 0.05g/kg per oral. gD= lycopene pigment 0.05g/kg per oral.
Throughout the experimental duration animals were deeply observed for variance in their conditions. They were sedated by ethanol and fixed on a dissecting panel. A vertical cut was given from sternum to pubic bone. Renal tissue was washed with saline water and fixed in 10% formalin for 24 hours, then kept for dehydration in 70% alcohol overnight. Further dehydrated by ascending concentrations of alcohol, after that cleared in xylene. Paraffin infiltration take place at 58 0 C in lab oven and paraffin blocks prepared. 4 micron dense slices excised on microtome & placed in warm H 2 O bath at 40 0 C and fixed on albumenized glass slides at 30 -32 0 C. Gomori's calcium phosphate was used to observe alkaline phosphatase activity in proximal convoluted tubular cells (8X ocular and 40X objective lens) [28].

Microscopic Observation of Renal
Tissue with the Help of Gomori's Calcium Phosphate Staining gA: Gomori's calcium phosphate stained renal sections of gA showed less or no black particles of alkaline phosphate in the apical area of cellular cytoplasm. (Fig.1).
gB: Gomori's calcium phosphate stained kidney sections showed marked deposition of dark black particles of alkaline phosphate in cellular cytoplasm of proximal convoluted tubules (Fig. 2). gC: Gomori's calcium phosphate stained section of this group showed rare deposition of black particles of alkaline phosphate in the cellular cytoplasm (Fig. 3).
gD: Gomori's calcium phosphate stained section of this group showed rare or no black particles of alkaline phosphate in the apical area of cellular cytoplasm just like gA. Gomori's calcium phosphate stained section of gC showed rare deposition of black particles of alkaline phosphate in the cellular cytoplasm at apical surface of proximal convoluted renal tubules. Similar results were also explained by [15,25] who determined that amendment & reversal of harmful effects in serum levels of alkaline phosphatase occur with administration of lycopene due to its potent antioxidant properties.

CONCLUSION
This experimental research study concluded that gB rats had marked accumulation of dark black particles of alkaline phosphate in cellular cytoplasm of proximal convoluted tubules due to renal injury on the other hand gC animals presented with rare deposition of black particles of alkaline phosphate in the cellular cytoplasm. Thus our hypothesis from this experimental work is that celebrex is injurious for kidneys& other vital organs while lycopene is advantageous for health because of its anti-oxidative properties.

LIMITATION OF STUDY
We have insufficient funds to do it in more depth.

CONSENT
It is not applicable.