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