NEPHROPROTECTIVE PLANTS

Moona A Latheef MadhukkalHH, Sithara Ravindran

National College of Pharmacy, Manassery, Calicut

INTRODUCTION

Man and his domesticated animals have since the time immemorial been largely dependent on

plants for the essential for their existence by way of food, clothing, shelter and medicines etc,

besides various other uses.1 Since disease, decay and death always coexisted with life, the study

of diseases and their treatment must have also been contemporaneous with the dawn of the

human intellect. The primitive man must have used as therapeutical agents and remedial

measures those things which he was able to procure most easily. There is no authentic record of

medicines used by the primitive man. But the Rigveda which is the oldest book in the library of

man supplies curious information on the subject.

In his work on plants and animal under domestication, Darwin says "From innumerable

experiments made through dire necessity by savages of every land, with the result handed down

by tradition, the nutritious, stimulating and medicinal properties of the most of unpromising

plants were probably first discovered." 

The doctrine of signatures would all account for the use of several plants as medicinal agents.

The reason for the extensive use of vegetable drugs may be the fact that plants are everywhere at

hand, their number is very great and their focus are distinct and peculiar and these are procured

without trouble.

It is greatly to the credit of people of India, that they were acquainted with a far large no. of

medicinal plants than the natives of any other country on the face of the earth.2 Many Indian

fruits, grains and vegetables employed as useful dietary articles forms a chief factor in the cure

of diseases, as well as preservation of health and good nutrition.3Herbs have always been the

principle form of medicine in India and they are becoming popular throughout the world, as

people strive to stay healthy in the face of chronic stress and pollution and to treat illness with

medicine that work in concert with the body's own defences. Thus medicinal plants play an

important role in the lives of rural people.4 A plant is said to be medicinal when "at least one part

possesses therapeutic properties."

One may recognize four stages in the development of the implements in the treatment of disease.

In the first stage, crude drug were employed, prepared in the roughest manner, such as powered

cinchona or metallic antimony. In the next stage, these were converted to more active and more

manageable forms, such as extractions or solutions, watery or alcoholic. In the third stage, the

pure active principles, separated from the crude drugs were employed Eg: morphine and quinine.

In the 4thstage, instead of attempting to extract out medicine from the natural products in which

they are contained, such substances are synthesized which possess particular desired

actions.2 Medicinal plants have curative properties due to the presence of various complex

chemical substances of different composition, which are found as secondary plant metabolites in

one or more parts of these plants.

The purpose of pharmaceuticals research is to develop new drugs. The discovery of medicinal

value of foxglove (Digitalis purpurea) is the case where traditional herbal knowledge led to

major advance in medicine. Phytochemical investigations on plants have not only yielded many

compounds of medicinal importance, but have also enriched our knowledge of the subject and

understanding of natural products.

Ayurveda and siddha system of medicine, the traditional heritage of India include many true

tested medicinal plants/drugs for various diseases and to which there is no answer in modern

medicine till today.

Nephrology is indeed an ancient discipline with a noble and distinguished legacy that spans at

least three previous millennia. A bronze artifact closely resembling the human kidney and dating

to 1300BC was excavated from the ruins of the temple of kition and at least thirteen references to

the kidney can be found in the old tastement. Before the time of the christ, Greek physicians

prescribed botanical material to promote diuresis and employed blood letting and other means

for removal of excess body fluids. Hyppocrates (460-375BC) was skilled in microscopic detail of

urine analysis. Artaeus of Cappadoicia (30-90AD) and Galan (130-200AD) recognized kidney as

the organ responsible for urine formation.5 By the middle of 1800, the structural complexity of

mammalian kidney was revealed & unraveled through improved optics & microscopy. If a few

names had to be chosen among the pioneers, we could mention Marcello Malpighi and Loreuzo

Bellin in Italy & Antoine Ferrein in France for the birth of renal anatomy, Sir William Bowman

in England & Karl Ludwig in Germany for renal physiology & Richard Bright in London &

Pierra Rayer in Paris for the diseases of kidney. Kidney is an important excretory organ in the

human body. The function of kidney is not only to excrete metabolic waste products, but also to

maintain the acid base balance, endocrine function like erythropoietin production.6

Ancient literature has prescribed various herbs for the cure of kidney disease. The term

"Pashanabeda" has been sited in literature to identify a group of plants, which have been

extensively used in indigenous system of medicine to dissolve urinary calculi & stones.

Eg: Aerva lanata, Crataeva nurvala, Pongamia prinnata etc. Some other plants mentioned in

literature include T.terrestris, O.sanctum, Zea mays etc.

ANATOMY & PHYSIOLOGY OF KIDNEY

Figure. 1: Section of Kidney

Paired kidneys are reddish bean shaped organs about 10-12cm long, 5-7cm wide, 3cm thick and

has a mass of 135-150g.10 The kidneys lie on the posterior abdominal wall, one on each side of

vertebral column, behind the peritoneum and below diaphragm. They extend from the level of

12th thoracic vertebrae to 3rd lumbar vertebrae.9 Near the centre of concave boarder is a deep

vertical fissure called the renal hilum, through which the ureter emerges from the kidney along

with blood vessels, lymphatic vessels & nerves.

The kidney consists of two distinct region, outer renal cortex & inner renal medulla. The urine

collects to calyx and then to renal pelvis which empties into ureter. The functional unit of kidney

is nephron and there are about 1million nephron in each kidney.10

STRUCTURE OF NEPHRON

Figure.2: Nephron

It consists of a tubule closed at one end, and the other end opening into a collecting tubule. The

closed end form Bowmann's capsule, which encloses the glomerulus.

The remaining parts of nephron is about 3cm long & consist of

Proximal convoluted tubule (PCT) Henley's loop Distal convoluted tubule

These nephrons are packed tightly to make up the kidney parenchyma.9

FUNCTIONS OF KIDNEY

The main function of kidney can be categorized as

Formation of urine Water & electrolyte balance Production of hormones & enzymes

In the resting adult, kidney receives 1.2-1.3litres of blood/min. In an adult, the GFR averages

120ml/min. The collecting duct of kidney is an area of fine control of ultrafiltrate composition &

volume, where final adjustment in electrolyte composition is made by the action of

mineralocortioid & ADH. The hypertonicity of medullary interstitium plays an important role in

concentrating the urine. The kidney not only excretes the metabolic substances, but also toxic

agents from the body.6 Hence kidney becomes one of the important targets for the toxicity of

agents more than other organs in body. Factors that make kidney particularly prone to actions of

nephrotoxicity include,

High levels of toxins are delivered to the kidney's large blood supply. The large surface area of renal tubular epithelium provide site for toxin interaction

& uptake. The availability of specific transport mechanisms that mediate cellular uptakes.

The normal concentrating mechanism of kidney can increase concentration of

toxins.

The presence of the metabolic processes in the renal tubular cell, can release toxic components &

induce damage. 

RENAL FAILURE

The term renal failure primarily denotes failure of the excretory function of kidney, leading to

the retention of nitrogenous waste products of metabolism in blood. In addition, there is failure

of regulation of fluid & electrolyte balance along with endocrine dysfunction.7 The renal failure

is fundamentally categorized into acute renal failure & chronic renal failure.

ACUTE RENAL FAILURE

Acute renal failure is characterized by azotemia that progresses rapidly over several hours or

days. It may or may not accompanied by oliguria & there is a sudden & reversible loss of renal

function.

Fig.3 Histopathology of kidney with ARF Fig.4. Histopathology of normal kidney

Early recognition of ARF is critical, because it is often asymptomatic. It is detected by

measuring serum creatinine level & is more specific than measurement of blood urea nitrogen

(BUN). There are many causes of ARF which could be,

Pre renal ARF 

It is due to under perfusion of kidney. It accounted for 21% of ARF cases. It can be thought of as

"a good kidney looking at a bad world." It is quickly reversible with appropriate therapy. 

Post renal ARF

It is caused by obstruction of urinary tract. It accounted for 10% of cases.

Intrinsic ARF

It is due to disease in parenchyma. It accounted for 69% of cases. Among the renal causes of

acute renal failure, acute tubular necrosis is more common accounting for 85% of incidence.

ATN occurs due to either ischaemia or toxins. The toxins can be either exogenous or

endogenous. The exogenous agents are radiocontrast agents, cyclosporins, antibiotics,

chemotherapeutic agents, organic solvents, acetaminophen, & illegal abortifacients.7

CHRONIC RENAL FAILURE

It is a syndrome characterized by progressive & irreversible deterioration of renal due to slow

destruction of renal parenchyma, eventually terminating in death when sufficient no. of nephrons

have been damaged.8 Various causes are glomerulonephritis, diabetes mellitus, chronic

pyelonephritis, hypertension.9 antineoplastic agents like cyclophosphamide, viniristne, cisplatin

etc.7

NEPHROTOXIC AGENTS

Drugs, diagnostic agents & chemical are well known to be nephrotoxic. The following are some

of the important nephrotoxic agents.11

A) Heavy metal 

Mercury, arsenic, lead, bismuth

B) Antineoplastic agents

Alkylating agents

Cisplatin, cyclophosphamide

Nitrosoureas: Streptozotocin, Carmustine, Lomustine & Semustine

Antimetabolites

High dose Methotrexate, Cytosine Arabinose, high dose 6-thioguanine, 5-flurouracil

Antitumour antibiotics

Mitomycin, Mithramycin, Doxorubicin

Biologic agents 

Recombinant leukocyte and interferon

C) Antimicrobial agents

Tetracycline, Acyclovir, Pentamidine, Sulphadiazine, Trimethoprin, Rifampicin

AmphotericinB 

D) Aminoglycosides

Gentamicin, Amikacin, Kanamycin, Streptomycin

E) Miscellaneous

Radiocontrast agents

Non-steroidal anti-inflammatory agents: Ibuprofen, Indomethacin, Aspirin etc

NEPHROPATHIESDUE TO TOXIC MECHANISM

Toxins may directly affect membrane permeability. Eg: with amphotericin & polyene antibiotics.

They can act by increasing the activity of membrane phospholipase & by inhibiting normal

reconstruction of the membrane. Phospholipid degradation products, lysophospholipids & free

fatty acids have membrane detergent properties. Even in the absence of major changes in

membrane permeability, the failure of plasma membrane pumps will cause potential injury

changes in the cation homeostasis of the cell eg: Na-K-ATPase & Ca ATPase pumps. The

activity of each may be affected by limitation of ATP, compromise of function of enzyme

protein or changes in the phospholipid microenvironment surrounding the enzyme.12 Toxin may

also lead to remodelling of the surface of the renal tubular cell, thus changing the area available

for transportation.

Numerous experiments have shown during cellular insult, an early and common change is the

accumulation of intracellular Calcium. This increase is found at plasma membrane, in

mitochondria and endoplasmic reticulum & in cytoplasm. An increase in intracellular Ca can

modify the permeability of internal membrane of the mitochondria & thus change in the

electrochemical gradient across it, which decreases the oxidative phopsphorylative capacity of

the mitochondria. Disordered permeability will then lead to loss of enzymes & nucleotides.12

In rats given gentamicin, the appearance of cellular necrosis & renal failure is well so related

with an increase in Ca in renal cortex & mitochondria. The intracellular metabolism of drugs

leads to the formation of reactive metabolites, which are toxic for cell, as are free radicals. The

superoxide ion normally formed, during oxidation forms hydroxyl radicals, which lead to lipid

peroxidation. This inturn causes oxidative deterioration of polyunsaturated lipids of membranes

& causes the dramatic modification of structure & function. The toxic agent reduces the

concentration of antioxidants, superoxide dismutase, glutathione, catalase, vit.E, ascorbic acid

which are the protective tissues that reacts & remove reactive oxygen species.4 Nephrotoxin

induced changes in tubule cells integrity may be sublethal or lethal. Such prelethal changes

include development of abnormally enlarged lysosomes & myeloid bodies, loss of brush border

membrane & vacuolization & dilation of the endoplasmic reticulum. Enzymuria resulting from

loss of some of these damaged membranes in the urine has been used to gauge the occurrence of

renal tubule cell injury & to follow it serially.11

TOXIC ACTIVATION AND FREE RADICAL PRODUCTION

Numerous in

vivo & in vitro studies have demonstrated the effect of free radicals like reactive oxygen

metabolites viz. superoxide, hydroxyl ions & hydrogen peroxide which are important mediators

of tissue injury. Free radicals can be defined as chemical species possessing unpaired electrons,

which are formed by hemolytic cleavage of a covalent bond of a molecule, by loss of a single

electron from a normal molecule or by the addition of a single electron to a normal molecule.

Free radicals may be positively charged (cation radical), negatively charged (anion radical) or

neutral These free radicals have very short half-life, high reactivity & damaging activity towards

macromolecules like proteins, DNA & lipids. These species may be either oxygen derived

reactive oxygen species (ROS) or nitrogen derived reactive nitrogen species (RNS). ROS

includes superoxide, hydroxyl, hydroperoxyl, peroxyl, alkoxyl as free radicals & hydrogen

peroxide, hypochlorous acid, ozone & singlet oxygen as non-radicals. The RNS are mainly nitric

oxide, peroxynitrile, & nitrogen dioxide & dinitrogen trioxide. Free radical injury & oxidative

stress have been implicated in many renal diseases like acute renal failure, IgA nephropathy,

anaemia of chronic renal failure & ischaemic kidney.

Superoxide ion & hydroxyl radical are formed during natural oxidative reaction by the action of

endoplasmic recticulum, mixed function oxidases, NADPH oxidases, xanthine oxidases etc. The

hydroxyl radical is highly reactive species formed by process of Fenton's reaction .The hydroxyl

radical can also be formed from oxidative metabolism of arachidonate by cyclooxygenase in the

presence of hydroperoxides.4

Many studies have shown that infusion of ROS from a chemical or cellular origin produces

glomerular dysfunction & injury to mesangial or endothelial cells with associated altered

golmerular permselectivity (protienuria). H2O2 perfusion via renal artery can produce

mesangiolysis & endothelial cell detachment.13

FORMATION OF FREE RADICAL

Free radical can be formed in three ways:

By hemolytic cleavage of covalent bond of a normal molecule, with each fragment

retaining one of the paired electrons. By loss of single electron from normal molecule. By addition of a single electron to a normal molecule.14They are constantly generated in

vivo.

PRODUCTION OF FREE RADICALS IN CELLS

Free radicals are produced by15

Ionising radiation. Accidental or deliberate Some enzymes utilize free radicals at their at their active sites in the process of catalysis.

Activated phagocytes also deliberately generate free radicals such as superoxide. These are produced by the leakage of electron transport chain such as those in

mitochondria & Endoplasmic reticulum. Metabolities of certain compounds exert toxicity through free radical mechanism

eg.Carbon tetra chloride is metabolized to toxic trichloromethyl free radicals by CYP

450 in liver.14

CISPLATIN TOXICITY

Cisplatin is a potent anticancer drug. It is used intensively in man, being effective in ovarian &

bladder carcinoma, neuroblastoma & head & neck carcinoma, & lymphoma as well as thyroid

endometrial neoplasm. However the most significant activity is observed in testicular cancer.

The clinical use of cisplatin is often complicated by nephrotoxicity Ototoxicity, gastrointestinal

disturbances like nausea, vomiting & myelosuppression. Early clinical trials of cisplatin in

cancer patients showed a striking incidence of persistent azotaemia & acute renal failure.

Cisplatin infusion at a dose of 20-mg/m2 over 4hr caused an increase in the filtration fraction &

decreased glomerular filtration rate. (offerman 1984). Nephrotoxicity is importantly modulated

as a result of biotrasformation. Tubular dysfunction has also been demonstrated very early after

cisplatin administration. The predominant pattern of injury include

Focal areas of tubular necrosis Desquamation of necrotic epithelial cells Necrotic cells with swollen mitochondria Cell membrane rupture with subsequent vesicle formation Necrotic debris filling the tubular lumens.

Loss of brush border Nuclear condensations.

Cisplatin has site-specific nephrotoxic effect on the proximal tubule of the rat. Three distinct

segments S1, S2, S3 been

described

for the proximal tubule of the rat by a number of investigators. A focal loss or thinning of the

microvillus

brush border was evident at all levels of microscopy. In some cells the brush border was

completely obliterated with only a few microvilli remaining. The cytoplasm of many cells

appeared condensed. Clumping of nuclear chromatin & increased number of cytoplasmic

vesicles could be seen in many of the injured cells. Other cells appeared to round up & lose their

normal orientation & often protruded into the tubular lumen. Completely necrotic cells were

evident & could be seen sloughing into the tubular lumen. In some areas, only a bare basal

lamina remained. Cells adjacent to these areas appeared to flatten out & send long thin

cytoplasmic process out to reline the basal membrane.

MECHANISM OF ACTION OF CISPLATIN INDUCED RENAL TOXICITY

The exact mechanism of cisplatin nephrotoxicity is unclear. Experimental studies have shown

that there is an abrupt fall in the effective renal plasma flow within 3 hrs of the i.p. dose of

cisplatin. It is known to be filtered by the glomeruli & concentrated in the glomerular filterate

from which it is activated in the presence of a low intra cellular chloride concentration. The low

intracellular concentration of chloride facilitates the displacement of chloride by the water

molecule yielding a positively charged, hydrated & hydroxylated complex. Hydration of

cisplatin induces formation of monochloro monoaquodiamino platin or diaquo diammineplatin.

These agents alkylate the purine & pyrimidine bases of nuclear material.12 Renal damage is seen

in proximal tubular S3 portion, the distal tubule & collecting duct. 

Other proposed explanation of the nephrotoxicity of cisplatin include the possibility that it

include generate reactive metabolites that bind covalently to tissue macromolecules. The

nephrotoxic effects might also be due to sulphydryl binding of heavy metal. A reduction in

sulphydryl groups in the rat renal cortex has been demonstrated; this occurred before any

significant change in renal function could be detected, suggesting that this biochemical change

may be a primary event. Cell fractionations have shown that the greatest decline of sulphydryl

groups occurs in the mitochondrial & cytosol fractions; these also had the highest concentrations

of platinum.12 A recent study found that cisplatin induced proximal tubule injury could be

ameliorated by the administration of hydroxylradical scavengers. In these studies cisplatin

(5mg/kg BW) caused lipid peroxidation. The hydroxyl radical scavenger prevented acute renal

failure by altering tubule damage & enhancing the regenerative response of damaged tubule cells

protection from cisplatin toxicity has generally focused on providing free radical scavengers.4

AMINOGLYCOSIDE INCLUDED NEPHROTOXICITY

Aminoglycoside antibiotics including gentamicin are widely used in the treatment of gram-

negative infections. However the major complication of the use of these drugs is nephrotoxicity,

accounting for 10-15% of all cases of acute renal failure. The nephrotoxicity of gentamicin is

well established in man & experimental animals. Renal tubular cell injury produced by

gentamicin evolves subacutely over several days & is characterized by following

Cellular necrosis Large lysosomes & myeloid bodies Mitochondrial structural alteration like swollen & ruptured mitochondria, Accumulation of gentamicin in renal proximal convoluted tubules. Suppression of free radical defence mechanism Phospholipidosis

The first step involved in the pathogenesis is the transport of the drug into proximal tubular cells

where they become concentrated & where they exert their toxic influence. The second step

involves the deleterious interaction of these agents with one or more intracellular metabolic

processes, which ultimately is expressed as a depression of renal function. With regard to

gentamicin, chronic exposure of cultured fibroblast to high levels of gentamicin, lead to

accumulation of the antibiotic within lysosomes accompanied by inhibition of lysosomal

sphingomylinase & a marked generalized phopholipidosis. The development of large lysosomes

& myeloid bodies during gentamicin nephrotoxicity has been well documented and inhibition of

kidney sphingomyelinase has been reported. Gentamicin induces inhibition of renal cortical

mitochondrial oxidative phosphorylation before histological evidence of severe proximal cellular

damage. There is significant reduction in whole kidney ATP levels, ADP dependant and

dinitrophenol (DNP) uncoupled respiration.16 Gentamicin treatment in vivo increased renal

cortical MDA levels decreased the total glutathione, increased the GSSH/GSH ratio, sharply

reduces levels of esterified arachidonic acid and induced a generalized shift from

polyunsaturated fattyacids. Gentamicin also decreased the activities of catalase & SOD.11

MECHANISM OF ACTION OF GENTAMICIN INDUCED RENAL TOXICITY

Gentamicin binds to the receptors located on the apical membrane of proximal tubular cells. It is

postulated that gentamicin being a cationic drug binds to the anionic phosphoinositides located

on the apical membrane. The binding of drug receptor is followed by pinocytosis of the drug-

receptor complex to a secondary lysosome. Within the lysosome, gentamicin might interfere with

the catabolism of receptor by directly inhibiting phospholipase C, by modifying substrate-

enzyme affinity or by raising the intralysosomal pH above the effective range of enzyme.

Inhibition of the activity of lysosomal phospholipase C leads to the accumulation of

phosphotidylinositol rich myeloid bodies within lysosomes leading to phospholipidosis.

AMELIORATION OF NEPHROTOXICITY

Antioxidant defence against ROS injury

Antioxidants are classified into 3 groups

Enzymatic antioxidants Vitamins Miscellaneous

Enzymatic antioxidants

Enzymes are the basic natural protectives of the organism against oxygentoxicity(burk80)

Eg: Superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase, G6PD.

Vitamins

These form another group of natural antioxidants

Eg: vit. A,C,E & K

Miscellaneous

Some other compound may also function as antioxidants 

Eg: Uric acid, beta-carotene, bilirubin, albumin, retinol, flavonoids and other phenolic

compounds of plant origin like curcumin dehydrozingerone, eugenol, isoeugenol.

An intricate system has evolved in respiring cells to prevent ROS causing injury. Two SODs are

known that catalyse dismutation of superoxide to hydrogen peroxide cytoplasmic (copper &

zinc) and mitochondrial (manganese) forms. Hydrogen peroxide is dealt with catalase containing

enzyme present in peroxisomes, and by cytoplasmic glutathione peroxidase, a seleno enzyme

which catalyses the reaction of glutathione disulphide. The glutathione disulphide is reduced

back to glutathione by an NADPH dependant enzyme, glutathione disulphide reductase.

A number of non-enzymatic defences against oxidant injury are available to the cell such as

alpha tocopherol, ascorbate, ceruloplasmin & the heat shock protein family. Melatonin, a pineal

hormone with antioxidant property, protects against Gentamicin-Induced nephrotoxicity. It

inhibits lipid peroxidation, restores the antioxidant levels & also regulates calcium channels.

The protective effect of sodium chloride has been shown in many experimental models of acute

toxic renal failure. For cisplatin ,glycerol, mercuric chloride & uranyl nitrate, a high-salt diet is

more protective than a normal diet. Sodium chloride may decrease the activity of the rennin-

angiotensin system.12

Diuretics have been also prescribed to prevent the appearance of acute renal failure. Mannitol

was the first to be used. It has been reported to ameliorate Amphotericin B & gentamicin

nephrotoxicity. It prevents an increase in urea & blood creatinine concentration, but does not

modify the intensity of the microscopic lesions.12

Flavanoids are phenolic compounds widely distributed in fruits, vegetables, plant extracts as well

as plant derived beverages Eg : tea & red wine. These have generated interest because of their

broad pharmacological effects such as vasoprotective, anti-inflammatory, antiviral & antifungal

actions. Many of these affects are related to their antioxidant properties, which may be due to

their ability to scavenge free radicals & to synergestic effects of other antioxidant. Another

mechanism not yet extensively studied, may result from interaction between flavanoid & metal

ions (esp iron & copper) leading to chelates. For Eg: it has been reported that concomitant

administration of quercetin with cisplatin showed considerable decreases in levels of marker for

nephrotoxicity & lipid peroxide & increased ATPase activities compared to CDDP treated group.

Glutathione content & antioxidant enzyme activities were significantly increased.

ANIMAL MODELS USED IN EXPERIMENTAL STUDIES

IN VIVO MODELS

GM treated albino rat Cisplatin treated albino rats Cisplatin treated rabbits GM treated guinea pigs Mercuric chloride treated mice Ethylene glycol treated mice

IN VITRO MODELS

Vero cells

PLANT SHOWING NEPHROPROTECTIVE ACTIVITY

1 EFFECT OF Aerva lanata ON GENTAMICIN & CISPLATIN MODELS OF ACUTE

RENAL FAILURE17

The ethanol extract of entire plant of Aerva lanata was studied for its nephroprotective activity in

cisplatin & gentamicin induced acute renal injury in albino rats of either sex. In the curative

regimen, the extract at dose levels of 75,150 & 300mg/kg showed dose dependant reduction in

the elevated blood urea and serum creatinine & normalized the histopathological changes in the

cisplatin model. In the gentamicin model, the rats in the preventive regimen also showed good

response to the ethanol extract at 300mg/kg. The findings suggest that the ethanol extract

of Aerva lanata possesses marked nephroprotective activity with minimal toxicity and could

offer a promising role in the treatment of acute renal failure caused by nephrotoxins like cisplatin

& gentamicin.

2 PROTECTIVE EFFECT OF Pongamia pinnata FLOWERS AGAINST CISPLATIN &

GENTAMICIN INDUCED NEHROTOXICITY IN RATS18

When ethanolic extract of flowers of Pongamia pinnata (300 &600mg/kg) was administered

orally in rats followed by cisplatin (5mg/kg ip), toxicity of cisplatin as measured by loss of body

weight, elevated blood urea & serum creatinine declined significantly. Similarly in gentamicin

(40mg/kg sc) induced renal injury, the extract 600mg/kg normalized the raised blood levels of

urea & serum creatinine levels. Reversal of cisplatin & gentamicin renal cell damage was

confirmed on histopathological examination. The results suggested that the protective effects is

through antioxidant property of two flavonoids kaempferol and 3,5,6,7,8-penta methoxy flavone.

3 Salviae radix EXTRACT PREVENTS CISPLATIN INDUCED ACUTE RENAL

FAILURE IN RABBITS19

The present study was carried out to determine if Salviae radix extract (SRE) exerts a beneficial

effect against cisplatin induced renalfailure in rabbits. Rabbits were pretreated with SRE orally

followed by cisplatin injection (5mg/kg ip). Cisplatin injection caused a reduction in GFR, which

was accompanied by an increase in serum creatinine levels. The fractional Na+ excretion and

lipid peroxidation were also increased. All these changes were prevented by SRE pretreatment.

Cisplatin treatment invitro in renal cortical slices increased LDH release and lipid peroxidation,

which were prevented by SRE and its effect may be attributed to its antioxidant action.

4 PROTECTIVE EFFECT OF GYCYRRHIZIN ON GENTAMICIN INDUCED ACUTE

RENAL FAILURE IN RATS20

The effects of glycyrrhizin (200 mg/kg/day) on renal function in association with the regulation

of aquaporin 2 water channel in rats with gentamicin (100 mg/kg/day)-induced acute renal

failure was investigated. Polyuria in rats with gentamicin-induced acute renal failure was

associated with down-regulation of renal aquaporin 2 in the inner and outer renal medulla, and

cortex. Glycyrrhizin administration restored the expression of aquaporin 2 with paralleled

changes in urine output. Changes in renal functional parameters, such as creatinine clearance,

urinary osmolality, and solute-free reabsorption, accompanying acute renal failure were also

partially restored after administration of glycyrrhizin. Histological changes in rats with

gentamicin-induced acute renal failure were also abrogated by glycyrrhizin treatment. The above

results suggest that glycyrrhizin treatment could ameliorate renal defects in rats with acute renal

failure induced by gentamicin.

5

Ginkgo biloba EXTRACT AMELIORATES GM INDUCED NEPHROTOXICITY IN

RATS21

The effect of Ginkgo biloba (EGb), a plant extract with an antioxidant effect, has been studied on

gentamicin-induced nephrotoxicity in male wistar rats. Ginkgo biloba extract (300 mg/kg BW)

was administered orally concurrently with gentamicin (80 mg/kg BW). Estimations of urine

creatinine, glucose, blood urea, serum creatinine, plasma and kidney tissue MDA were carried

out after gentamicin treatment. Kidneys were examined using histological techniques. Blood

urea and serum creatinine were increased with gentamicin. Creatinine clearance was significantly

decreased with gentamicin. Changes in blood urea, serum creatinine and creatinine clearance

induced by gentamicin were significantly prevented by Ginkgo biloba extract. There was a rise

in plasma and kidney tissue MDA with gentamicin, which were significantly reduced to normal

with Ginkgo biloba extract. Histomorphology showed necrosis and desquamation of tubular

epithelial cells in renal cortex with gentamicin, while it was normal with Ginkgo biloba extract.

These data suggest that supplementation of Ginkgo biloba extract may be helpful to reduce

gentamicin nephrotoxicity.

6 EFFECT OF Cassia auriculata ROOT EXTRACT ON CISPLATIN & GM INDUCED

RENAL INJURY22

The ethanol extract of the roots of Cassia auriculata was studied for its nephroprotective activity

in cisplatin- and gentamicin-induced renal injury in male albino rats. In the cisplatin model, the

extract at doses of 300 and 600 mg/kg body wt. reduced elevated blood urea and serum

creatinine and normalized the histopathological changes in the curative regimen. In the

gentamicin model, the ethanol extract at a dose of 600 mg/kg body wt. reduced blood urea and

serum creatinine effectively in both the curative and the preventive regimen. The extract had a

marked nitric oxide free-radical-scavenging effect. The findings suggest that the probable

mechanism of nephroprotection by C.auriculata against cisplatin- and gentamicin-induced renal

injury could be due to its antioxidant and free-radical-scavenging property.

7 AGED GARLIC EXTRACT ATTENUATES GM INDUCED RENAL DAMAGE AND

OXIDATIVE STRESS IN RATS23

Aged garlic extract (AGE), an antioxidant, has a protective role in this experimental model of

male Wistar rats were studied. AGE was given at a dose of (1.2 mL/kg/12 hours) followed by

GM (70 mg/kg/12 hours). Nephrotoxicity was made evident by:

1) the increase in blood urea nitrogen and plasma creatinine

2) the decrease in plasma glutathione peroxidase (GPx) activity and the urinary increase in N-

acetyl-beta-D-glucosaminidase activity and total protein

3) necrosis of proximal tubular cells 

4)increase in the renal levels of oxidative stress markers: nitrotyrosine and protein carbonyl

groups and the decrease in manganese superoxide dismutase (Mn-SOD), GPx, and glutathione

reductase (GR) activities. 

These alterations were prevented or ameliorated by AGE treatment. Furthermore, AGE

prevented the GM-induced The protective effect of AGE was associated with the decrease in the

oxidative stress and the preservation of Mn-SOD, GPx, and GR activities in renal cortex. These

data suggest that AGE may be a useful agent for the prevention of GM-nephrotoxicity.

8 THE EFFECTS OF Nigella sativa OIL ON GM NEPHROTOXICITY IN RATS24

In this work, tested whether oral treatment of rats with N. sativa oil (0.5, 1.0 or 2.0 ml/kg/day)

would ameliorate nephrotoxicity of GM (80 mg/kg/day im) concomitantly with the oil.

Nephrotoxicity was evaluated histopathologically and by measurement of concentrations of urea,

creatinine and total antioxidant status (TAS) in plasma and reduced glutathione (GSH) and TAS

in kidney cortex. The results indicated that GM treatment caused moderate proximal tubular

damage, significantly increased the concentrations of creatinine and urea, and decreased that of

TAS and GSH. Treatment with N. sativa oil produced a dose-dependent amelioration of the

biochemical and histological indices of GM nephrotoxicity that was significant at the two higher

doses used, and it increased GSH and TAS concentrations in renal cortex and enhanced growth.

The results suggest that N. sativa may be useful in ameliorating signs of GM nephrotoxicity in

rats.

9 FLAVONOID OF Drynaria fortunei PROTECTS AGAINST ARF25

The flavonoid fraction (FF) from Drynaria fortunei was investigated to determine its biological

activity expression in three acute renal failure animal models Guinea pigs & mercuric chloride

treated mice. Guinea pigs received 100 mg/kg of gentamicin & 10 mg/kg of FF. FF treatment

prevented the GM toxicity, ie; the increase in BUN and creatinine levels. Mice were treated once

with 6 mg/kg of mercuric chloride, followed by 10 mg/kg of FF. BUN and creatinine levels were

found to be significantly higher on the mercuric chloride treatment and is ameliorated by FF

treatment. In conclusion, the present study suggests that FF prevents nephrotoxicity, improves

kidney function and promotes kidney primary epithelial tubular cell regeneration. 

10

THE ROLE OF GINSENOID-Rd IN CISPLATIN INDUCED ARF26

Ginsenoside-Rd has been proved to decrease the severity of renal injury induced by cisplatin, in

which proximal urinaferous tubules represent the main site of injury. When ginsenoside-Rd was

given orally at a dose of 1 or 5 mg/kg body weight/day prior to cisplatin injection, the activities

of the antioxidation enzymes superoxide dismutase and catalase were higher, while

malondialdehyde levels in serum and renal tissue were lower in the treated rats than in the

controls. The levels of urea nitrogen and creatinine in serum were decreased in rats given

ginsenoside-Rd. Decreased urinary levels of glucose, sodium and potassium reflected a

protective action against the renal dysfunction caused by cisplatin. In addition, it was

demonstrated that ginsenoside-Rd affected cultured proximal tubule cells exposed to cisplatin.

11 NEPHROPROTECTIVE ACTION OF Tribulus terrestrisAND Crataeva nurvala IN

ALBINO RATS27

Nephrotoxic model was developed in male albino rats by administering GM. The aqueous extract

of fruits of T.terrestris (65 or 130mg/kg) and C.nurvala (70 or 145mg/kg) after GM

administration. Urine was examined for sugar, albumin, RBC & epithelial cells.

Histopathological changes were also noted. The drug showed a dose dependant nephroprotective

action against GM toxicity. The results indicate that the two indigenous plants would ameliorate

renal effects in albino rats with acute renal failure induced by GM.

12 EFFECT OF Ocimum sanctum AQUEOUS LEAF EXTRACT ON GM INDUCED

NEPHROTOXICITY IN RATS28

Nephrotoxicity was induced in rats by GM (180mg/kg/day ip).O.sanctum aqueous leaf extract

(OS) was given orally at a dose of 100 mg/kg/day along with GM. Concurrent administration of

OS significantly prevented rise in levels of serum creatinine, blood urea & plasma MDA which

elevated by GM. It also significantly prevented the histological damage caused by GM. The

results suggested that OS probably by virtue of its antioxidant property prevented GM induced

nephrotoxicity in rats.

13 THE EFFECT OF TREATMENT WITH THE MEDICINAL PLANT Rhazya stricta ON

GM INDUCED NEPHROTOXICITY29

Crude water extract of R. Stricta leaves (0.25, 0.5 and 1 g/Kg) was given orally to rats and

thereafter, concomitantly with GM (80 mg/Kg/day). Nephrotoxicity was evaluated

histopathologically and biochemically by measuring the concentrations of urea and creatinine in

serum, reduced glutathione (GSH), lipid peroxidation and superoxide dismutase (SOD) activity

in kidney cortex. The results suggested that a dose-related amelioration in the indices of toxicity

was noted when the two higher doses of the plant extract were given. The two higher doses,

significantly and dose-dependently increased SOD activity and GSH concentration, and

decreased that of lipid peroxides in the kidney cortex. These results suggest that R. stricta water

extract may contain compounds that could potentially ameliorate GM nephrotoxicity in rats.

14 RENOPROTECTIVE EFFECT OF GRAPE SEED EXTACT IN ETHYLENE

GLYCOL INDUCED NEPHROTOXIC MICE30

Grape seed extract in ethylene glycol (EG) induced nephrotoxicity in mice was studied for its

nephroprotective activity. Mice received grape seed extract 100mg/kg BW was given after EG

(2ml/kg BW po) administration. Grape seed extract in mice produced significant reduction of

urinary LDH, blood urea, creatinine & dilated tubules lined by normal intact epithelium

indicating recovery. The results suggest that the renoprotective effect of Vitis vinifera seed

extract is due to improvement in antioxidant status.

15

CYTOPROTECTIVE ROLE OF Solanum nigrum AGAINST GM INDUCED

KIDNEYCELL (VERO CELL) DAMAGE INVITRO31

The 50% ethanol extract of the whole plant of Solanum nigrum was tested in vitro for its

cytoprotection against gentamicin-induced toxicity on Vero cells. Cytotoxicity was significantly

inhibited as assessed by the Trypan blue exclusion assay and mitochondrial dehydrogenase

activity (MTT) assay. The test extract also exhibited significant hydroxyl radical scavenging

potential, thus suggesting its probable mechanism of cytoprotection.

16 EVALUATION OF NEPHROPROTECTIVE EFFECT OF INDIAN MEDICINAL

PLANS (IMPs) IN EXPERIMENTAL GM INDUCED NEPHROTPXICITY32

The effect of administration of IMPs, Withania somnifera, Emblica officinalis, Glycyrrhiza

glabra on BUN, serum creatinine, bodyweight MDA, renal histopathology were evaluated with

administration of GM (150mg/kg/day) in female rats. Concurrent administration of IMPs &

alpha lipoic acid prevented the rise in BUN, serum creatinine, kidney MDA to varying degrees.

Thus IMPs show promise as protective agents against experimental nephrotoxicity.

DISCUSSION

This study was conducted to establish the nephroprotective activity of plants. Various models

have been used to substantiate the nephroprotective activity of herbals. They were GM in albino

rats, cisplatin in rabbits, mercuric chloride in mice, ethylene glycol in mice etc. These

nephrotoxic agents caused nephropathy mainly due to their free radical generation in kidney

tissues. And the kidney damage was indicated by changes in renal function parameters like

creatinine, BUN, and the enzymes suchn as GPx, SOD and was also confirmed

histopathologically. Above works certified that, by ameliorating all the allied effects, mainly due

to antioxidant property the plants likeA.lanata, P.pinnata, C.auriculata, S.radix, G.glabra,

G.biloba, N.sativa, D.fortunei, T.terrestris, C.nurvala, O.sanctum, S.nigrum, V.vinifera have

nephroprotective activity.

CONCLUSION

As we gone through various studies on treatment of kidney disorders, we can conclude that

herbal plants play a unique role in medicine. There is no synthetic drug which relieve overall

insufficiency of kidney. But indigenous plants possess tissue rejuvenator property which is

anyway unavoidable. To Indians, who are brought upon Indian food, soul & climate with Indian

habits of life and environment, Indian drugs naturally suit better and safer than European

constitution built upon their peculiar food, climate, habits and manner of life. This may perhaps

be the reason why in numerous cases,where synthetic medicines fails, Indiginous system of

medication succeed.

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Posted by Soman   at 11:49 PMLabels: AMELIORATION, ANATOMY PHYSIOLOGY OF KIDNEY, CISPLATIN INDUCED RENAL TOXICITY, FLAVONOID OF Drynaria fortunei, FORMATION OF FREE RADICAL, GENTAMICIN, nephroprotective plants