INTRODUCTION:

The term acute renal failure (ARF) is currently substituted by acute kidney injury (AKI). AKI is a reversible condition in which there is a sudden decline in renal function, manifested by hourly/daily/weekly elevation in serum creatinine and blood urea nitrogen (BUN). Acute renal failure (ARF), characterized by sudden loss of the ability of the kidneys to excrete wastes, concentrate urine, conserve electrolytes, and maintain fluid balance, is a frequent clinical problem, particularly in the intensive care unit, where it is associated with a mortality of between 50% and 80%.¹

Gentamicin toxicity:

Gentamicin is an aminoglycoside antibiotic that is still commonly used in the treatment of life-threatening in- fections. Their broad-spectrum activity against aero- bic gram +ve and gram-ve organisms, their chemical stability, and their rapid bactericidal action has often made them first-line drugs in a variety of clinical situ- ations.^2,3

However, high concentrations of these antibiotics are nephrotoxic. In some cases, this side effect is so severe that the use of the drug must be discontinued. It has been estimated that up to 30% of patients treated with aminoglycosides for more than 7 days show some signs of nephrotoxiciy.⁴

About 15-30% of treated subjects with these drugs are susceptible to gentamicin toxicity which manifested clinically as non-oligo uric renal failure, with a slow rise in serum creatinine and hypo osmolar urinary output develops after the prolong consumption or a time consuming treatment, produces by them due to this reason they have major limitations in clinical use.⁵

Nephroprotective Agent:

Ancient studies showed the nephroprotective activity of various herbs which are used for the treatment of kidney disease. There are various medicinal plants which posses the curative properties due to the presence of various complex chemical substances are used for the nephroprotective activity. In table No 1, the agent which are active against Nephrotoxicity produces by several drugs. They are known as Nephroprotective agents because they shows protective activity against nephrotoxicity.

Table-1 Description of medicinal plant possessing Nephroprotective activity.

Botanical Name	Family	Part used	Chemical constituents	Screening method	References
Strychnos potatorum	Loganiacseae	Seed	Flavanoids, Phenols, Saponins, Alkaloids, Steroids, Tannins, Glycosides, and Lignins.	Gentamycin induced	6
Aerva javanica	Amaranthaceae	Fresh roots	Isoquercetin, 5 methylmellein, 2-hydroxy-3-O-β-primeverosidenaphthalene-1,4-dione, Apigenin 7-Oglucoronideand Kaempferol	Cisplatin induced	7
Carica papaya	Caricaceae	Seed	Flavanoids, Phenols, Alkaloids, Protein, Sterols, Terpenoids, Carbohydrates, Steroids, Tannins, Glycosides, Terpins and Saponins	Cisplatin induced	8
Ficus religiosa L	Moraceae	Latex	Flavonoids, Amino acids and Tannins	Cisplatin induced	9
Crataeva nurvula	capparidaceae	Fruit	Kaemferol-3-O-a-D-glucosideFlavanoids, Glucosinolates, Steroids, Lupeol and Tannins	Gentamycin induced	10
Orthosiphon stamineus	Laminecawe	Whole plant	Flavanoids, Phenols, Carbohydrates, Steroids, Tannins	Gentamycin induced	11
Aerva lanata	Amaranthaceae	Whole plant	Botulin, β-sitosterol, Amyrin, HentriacontanStigma sterol	Gentamycin induced	12
Pedalium murex Linn	Pedaliaceae	Dried fruits	Flavanoids, Flavones, Alkaloids, Triterpenoids	Cisplatin induced	13
Vernonia cinerea	Compositae	Aerial parts	Triterpenoids like α-amyrin, β-amyrinand lupeol.	Cisplatin induced	14

LUTEOLIN:

Luteolin is one of the most common flavonoids present in edible plants and in plants used in traditional medicine to treat a wide variety of pathologies. Luteolin is widely distributed in the plant kingdom.

MATERIAL AND METHODS:

The study was initiated after getting IAEC (Institutional Animal Ethics Committee) approval.

Chemicals and Reagents:

Luteolin, gentamicin, normal saline and 10% formalin were purchased from CDH, Mumbai, India. All chemicals used were off analytical grade.

Animals:

Healthy adult wistar albino rats of between 2 to 3 months of age weighing about 150 gm to 250 gm were used for the study. All the animals were housed in poly propylene cages, maintain under the standard conditions (12h light: 12h dark cycle; 25+ 30 C; 35-60% humidity). They were fed with standard rat pellet diet (Hindustan lever Ltd., Mumbai India) and water.

Experimental procedure:

Luteolin was administered at the dose of 100mg/kg and 200mg/kg was selected as par Galvez et al.,1997to the rats weighing between 150 and 200 g in groups of four were used (n =4) each group consisting 5 rats. ¹⁵

In vivo Nephroprotective Activity:

Induction of Experimental Nephrotoxicity:

Gentamicin (80 mg/kg bw) solution were prepared separately in sterile distilled water. Rats were treated with gentamicin for 10 days by oral route. In order to study the effect of luteolin rats 100 mg/kg and 200mg/kg bw were used respectively. Rats were divided into four groups as following protocol.

Treatment protocol:

GROUP I: Normal control (n =6, the animal were given normal saline only)

GROUP II: Nephrotoxic control (n=6 the animal were given gentamicin for 10 days)

GROUP III: Treatment group (n=6 the animal were given gentamicin + luteolin (100 mg/kg bw) for 10 days)

GROUP IV: Treatment group (n=6 the animal were given gentamicin + luteolin (200 mg/kg bw) for 10 days)

Rats were treated as per the treatment protocol. Body weights of these rats were monitored sequentially in control and experimental animals for a period of 10 days.

Biochemical estimation:

Blood sample were collected via retro- orbital puncture at the end, the serum was rapidly separated and processed for determination of creatinine, serum urea, blood urea nitrogen (BUN), using of span Diagnostic kits. Body weight of animal was also recorded.

Histopathological Studies:

Rats were sacrificed and both kidneys were isolated from each rat. The kidneys wereprocessed for Histopathological examination. The kidney were excised quickly and fixed in 10 % formalin and stained with haemotoxylin and eosin and then observed under microscope for degeneration, fatty changes, necrotic changes and evidence of nephrotoxicity if any.

Statistical Analysis:

The results were expressed as mean + SEM. Statistical analysis was carried out by using Oneway ANOVA followed by Dunnet’s ’t’ test and P< P<0.05, **P<0.01, ***P<0.001 was considered significant.

RESULT:

Nephroprotective activity:

The results of nephroprotective activity of Luteolin on Gentamicin treated rats are shown in fig. 3.1-3.4. The urine creatinine, serum urea and blood urea nitrogen increased in gentamicin treated animals when compared to control. The luteolin treatment significantly reversed the level of urine creatinine, serum urea and blood urea nitrogen when compared to gentamicin treated rats (Figure 3.1-figure 3.4)

Table3.1 Nephroprotective activity of luteolin in gentamicin induced nephrotoxicity in Rats

GROUP	Serum Creatinine (mg/dl)	Serum urea Mg/dl	Urine Creatinine (mg/dl)	BUN Mg/dl	Body weight (% change)
Control	0.63+ 0.15	43.15+2.15	95.97+4.34	18.87+1.67	2.79+0.43
Gentamicin 80mg/kg	3.21+0.21	176.42+2.45	285.31+5.15	82.45+1.45	-11.98+0.36
Gentamicin 80mg/kg+ Luteolin 100mg/kg	2.08+0.10*	79.61+2.34*	135.83+5.55*	34.92+1.58*	-8.54+0.55
Gentamicin 80mg/kg+ Luteolin 200mg/kg	1.60+0.12**	54.26+3.98***	96.41+4.37 **	21.88+1.73*	-6.21+0.46

n=6, values are expressed mean + S.E.M.*P<0.05, **P<0.01, ***P<0.001 when compared with control group and gentamicin treated group.

Figure 3.1: Effect of Luteolinadministration on serum creatinine in rats

n=6, values are expressed mean + S.E.M.*P<0.05, **P<0.01, ***P<0.001 when compared with control group and gentamicin treated group.

Figure 3.2: Effect of Luteolinadministration on serum urea, blood urea nitrogen and urine creatinine in rats

n=6, values are expressed mean + S.E.M.*P<0.05, **P<0.01, ***P<0.001 when compared with control group and gentamicin treated group.

Figure 3.3: Effect of Luteolinadministration on percent change in body weight and kidney weight in rats

n=6, values are expressed mean + S.E.M.*P<0.05, **P<0.01, ***P<0.001 when compared with control group and gentamicin treated group.

(a)

(b)

(c)

(d)

Fig 3.4 Histopathological view of renal section from different groups of animal

Evidence for the protective effect of Luteolin in rats treated with gentamicin;(a) control,

(b) toxicant, (c) Luteolin at the dose of 100 mg/kg(d) Luteolin at the dose of 200mg/kg.

DISCUSSION:

Nephrotoxicity (from Greek: nephrons, "kidney") is a toxic effect of some substances, such as endogenous and exogenous compound and several drugs on the kidney. Nephrotoxicity can be estimated by a simple blood test. An increased level of creatinine shows declained renal function. Nephrotoxicity is caused by the drugs such as several groups of antibiotics. According to site and mechanism the nephrotoxicity can be classified such as interstitial nephritis, renal tubular necrosis, and crystalurea. In Nephrotoxicity there is increase amount of plasma drug level and decreased amount of urine drug concentrations and this leading to greater toxicity, the forced diuresis can also be used for the protection of kidney. Free radicals and ROS are also responsible to produce oxidative stress on kidneys.

When there is a expose of Nephrotoxic drugs, substance or the toxins it causes nephrotoxicity. There is number of therapeutic substance which can adversely affect the kidney causes acute renal failure like aminoglycosides antibiotics, chemotherapeutic agent and other potent drugs causes chronic interstitial nephritis and nephritic syndrome.¹³ In the present study, nephrotoxicity

In rats was successfully produced by administrating gentamicin at the dose of 80mg/kg orally. It affects the defense system by increasing the production of ROS which cause inactivation of antioxidant enzymes. The impairment of kidney function is accompained by increased in serum creatinine, serum urea, urea creatinine and blood urea nitrogen level that indicates lipid peroxidation. It is one of the essential compound for maintaining cell integrity participation in the cell metabolism. The significant and progressive weight loss in gentamicin treated rats possibly be due to the injury renal tubules and the subsequent loss of tubular cells to reabsorbed water, leading to dehydration and loss of body weight.

Luteolin is an antioxidant, active against the effect produced by the ROS at the renal tubules or kidney cells. When concurrent administration of luteolin with gentamicin in rats the biochemical parameters were reduces that showed the activity of luteolin against the nephrotoxicity.

In this study the result suggest that the statistically significant difference in biochemical parameters in toxic control group, indicate that nephrotoxicity has been induced by gentamicin. Following treatment with the luteolin all the parameters were reduced and restored to normal values.

CONCLUSION:

The present study of Luteolin is found to have nephroprotective effect in gentamycin-induced acute renal failure in Wistar albino rats. Luteolin is found to have significant action in nephrotoxicity in albino rats.

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Received on 05.07.2019 Modified on 31.07.2019

Res. J. Pharma. Dosage Forms and Tech.2019; 11(3):181-185.

DOI: 10.5958/0975-4377.2019.00032.6