Santhosh Suddagoni, Int. J. in Pharm. Sci.

, 2023, Vol 1, Issue 6, 86-98 | Research

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PHARMACEUTICAL SCIENCES

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Research Article
Nephroprotective and Anti-Inflammatory Activity of Combination of
Punica Granatum (Fruit Peel) And Tectona Grandis (Bark)
Dr. Santhosh Suddagoni1*, Dr. kapil malviya2, Dr. Santikari Sesha Phanindra3, Dr.
M. Venkata Ramana4, Bodige Divya5, Dr. Ganesh Akula6
1
Department of pharmacology, Mansarovar Global University, Bilkisganj, Dist. Sehore, Bhopal, Madhya
Pradesh, India 466001.
2
Department of pharmacology, Mansarovar Global University, Bilkisganj, Dist. Sehore, Bhopal, Madhya
Pradesh, India 466001
3
Department of pharmaceutical chemistry, Surabhi Dayakar Rao College of Pharmacy, Rimmanaguda, Gajwel,
Siddipet, Telangana, India-502312
4
Department of pharmaceutical chemistry Surabhi Dayakar Rao College of Pharmacy, Rimmanaguda, Gajwel,
Siddipet, Telangana, India-502312
5
Department of pharmacology, Jyothismathi intitute of pharmaceutical sciences, Ramakrishna colony,
thimmapur, karimnagar, Telanagana, india-505481
6
Department of pharmaceutical chemistry, CMR college of pharmacy. Kandla koya, Hyderabad, Telangana,
india-501401.

ARTICLE INFO ABSTRACT

Received: 30 May 2023 Herbal medication in addition referred to as herbal treatment or
Accepted: 02 June 2023
phytoconstuients refers to employ a plant’s seed, berries, roots, leaves, bark or
Published: 14 June 2023
flowers for healthful functions. The procedure of herbs to treat a variety of
Keywords:
Phytomedicine, Punica
different ailments is universal and exists in every human culture on Earth. The
granatum, Tectona grandis, plants Punica granatum and Tectona grandis is important medicinally and
Diabetes, Nephroprotective and claimed to be helpful in the treatment of different ailments like diabetes,
anti-inflammatory actions inflammations, eruptions and bites of poisonous animals etc., and contains
DOI: different classes of compounds of pharmacological importance such as,
10.5281/zenodo.8039301 alkaloids, glycosides, saponins, flavonoids and triterpines and have been screen
biologically for Nephroprotective and anti-inflammatory actions.

*Corresponding Author: Dr. Santhosh Suddagoni

Address: Department of pharmacology, Mansarovar Global University, Bilkisganj, Dist. Sehore, Bhopal, Madhya Pradesh,
India 466001
Email : [email protected]
Relevant conflicts of interest/financial disclosures: The authors declare that the research was conducted in the absence of
any commercial or financial relationships that could be construed as a potential conflict of interest.

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INTRODUCTION compounds of pharmacological importance such
Herbal medication in addition referred to as herbal as, alkaloids, glycosides, saponins, flavonoids and
treatment or phytomedicine refers to, berries, triterpines and have been screen biologically for
roots, leaves, bark or flowers for healthful Nephroprotective and anti-inflammatory actions15-
functions1-5. Herbalist contains a long tradition of 18
.
use outside of typical medication. Ancient Chinese
and Egyptian papyrus writings describe healthy
use for plants at the same as early as 3000 BC3.
Native cultures (such as African and Native
American) use herbs in their heal rituals, whereas
others developed ancient healthy systems (such as
written material and ancient Chinese medicine)
Fig. No.: 2 Fruit peel of Punica granatum
during which seasoned therapy were used.
Researchers start that folks in numerous elements
of the globe attend uses of similar plants for a
similar purpose6-9. The procedure of herbs to treat
a variety of different ailments is universal and
exists in every human culture on Earth. Despite
this, the major use of medical herbs still occurs in
societies which are not fully developed Because of
the high costs involved with industrialized modern Fig. No.: 3 Tectona grandis plant
medicines, many people living in increasing
nations simply do not have the financial resources METERIALS AND METHOD
to give for them, and as a result, they are enforced Table No: 1 List of materials and equipment’s used
to use normal herbs as an affordable alternative.10- during experiment.
14
. Sr. No. Name of the materials and equipment
Anesthetic ether (Sigma Solvents and
1
Pharmaceuticals, Mumbai)
2 Formalin (Hi-Media Laboratory, Mumbai.)
3 Ethanol (Nice-Cochin)
4 Carrageenin (Nice-Cochin)
5 Diclofenac sodium (Cipla, Mumbai)
6 Distilled water.
7 Evans blue (S.D. Fine chemicals, Mumbai)
Lead Mercury (S.D. Fine chemicals,
Fig.No.1 Herbs 8
Mumbai)
The plants Punica granatum and Tectona grandis Restraint cages (Shri. Venkateswara
9
is important medicinally and claimed to be helpful Enterprises, Bangalore)
in the treatment of different ailments like diabetes, 10 Chloroform (Nice-Cochin)
inflammations, eruptions and bites of poisonous Refrigerator (Whirlpool-Kelvinator India
11
animals etc., and contains different classes of Ltd, India)

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Collection and authentification of plant Experimental Procedure:
materials: Albino rats (150-200 g) be separated into 6 groups
The stem of Punica granatum and Tectona each containing 6 animals they be fast overnight
grandis were collected from limited area and prior to and through the test but include free access
authentified by botanist Dr. Narasingham BSI. to water. Group A be served as toxicant control
Preparation of special extracts (petrolium treated among toxicant Carrageenan, group B with
ether, chloroform, ethanolic and aqueous Diclofenac (40 mg/kg p.o.) that serve because
extracts): normal Groups C, D, E and F be administer with
The fruit peel of Punica granatum and bark of EEPG, EETG and AEPG, AETG (200 mg/kg and
Tectona grandis were dried in shade at room high 400 mg/kg dose p.o) respectively. The rats of
temperature then Subjected to size decrease to a groups B, C, D, E and F be administer with 1% of
fine powder with the help of mixer grinder. Then Carrageenin into sub plantar part of right hind paw
the powder was extract with petroleum ether, of rats 1h after administration of
chloroform, ethanol and water consecutively by Diclofenac/extracts. directly here later than the
simple extraction. The extract was move into the oedema volumes of the injected paws be
before weighed clear beaker and evaporate to a considered plethysmographically at prefixed time
thick paste on the water bath, maintained at 50oC interval. The variation among paw volume of the
to get extracts. The extract be lastly air dry treat animals is considered and the mean oedema
thoroughly to remove all traces of the solvent and quantity be calculated19-28. Percentage decrease
the percentage yield was planned each time before in oedema volume was calculated by use the
extracting with next solvent, marc will be dried in formula.
hot air oven below 50oC and each extract will be Vo - Vt
concentrated by distilling off the solvent and then Percentage reduction = x 100
Vo
evaporated to dryness on a water bath to get the
Where,
extracts.
Vo = Volume of the paw of control at time ‘t’.
Pharmacological evaluation:
Vt = Volume of the paw of drug treated at time ‘t’.
Depending upon the occurrence of
Statistical analysis:
phytoconstituent in the different extracts, the
All consequences will be expressed as mean ±
ethanolic and aqueous extracts are selected for the
SEM from 6 animals. Statistical variation in mean
following pharmacological actions.
will be analyze use one-way ANOVA (analysis of
• Anti-inflammatory activity variance) follow by position hoc test (Dunnett‘s‘t’
• Nephroprotective activity test). P< 0.05*, 0.01** and 0.001*** will be
Anti-Inflammatory Activity By Carrageenan measured as statistically important29-37.
Induced Paw Oedema60, 61,74 Nephroprotective activity by Gentamicin
Group A: Control (Carrageenin 1%) Induced Nephrotoxicity in Rats47, 52
Group B: Standard (Diclofenac 30 mg/ kg) The evaluation of the ethanolic extract for
Group C: EEPG+EETG (200 mg/kg p.o) nephroprotective action be done according to the
Group D: EEPG+EETG (400 mg/kg p.o) process known in the literature with minor
Group E: AEPG+AETG (200 mg/kg p.o) modification.
Group F: AEPG+AETG (400 mg/kg p.o)

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Total 36 animals are in use and 6 rats were selected Preliminary phytochemical screening:
in each of the follow groups: EEPG+EETG and AEPG+AETG were subjected
Group I: Control group for phytochemical selection and start to include
Group II: Gentamicin control group (60mg/kg) tannins, sterols, flavonoids, glycosides, and
Group III: Gentamicin + Ethanolic extract alkaloids triterpines in ethanolic and aqueous
(400mg/kg) extracts. The phytochemical constituents of
Group IV: Gentamicin + ethanolic extract different extract of Punica granatum and Tectona
(200mg/kg) grandis are shown below:
Group V: Gentamicin + Aqueous extract Table No.:3. Phytochemical evaluation of
(400mg/kg) combined extracts of Punica granatum and Tectona
Group IV: Gentamicin + Aqueous extract grandis
(200mg/kg). Sr. Petroleu Chloro Etha
Tests Water
No. mether form nol
Activity profile of the test formulations in
01 Alkaloids -ve -ve +ve +ve
gentamicin induced changes in different
02 Carbohydrates -ve -ve +ve +ve
parameters:
Serum uric acid is the ending product of purine 03 Flavonoids -ve -ve +ve +ve
catabolism. So, any fault in the glomerular 04 Saponins -ve -ve +ve +ve
filtration rate causes the rise into the stage of uric 05 Sterols +ve +ve +ve +ve
acid in the blood. The raise later than gentamicin 06 Tannins -ve -ve +ve +ve
07 Glycosides -ve -ve +ve +ve
can be credited to the GFR impairment. The
exchange of the elevation by any substance may be
analytic of the exchange of the GFR Pharmacological activities:
impairment53-59. Acute oral toxicity study:
RESULTS AND DISCUSSION The mice treated with EEPG+EETG and
The nature and percentage yield of different AEPG+AETG at a dose of 200 mg/kg, p.o.
extract be given below: exhibited usual behaviour, without any symbols of
Table No.: 2 Nature and Percentage yield of the passivity, stereotypy and vocalization. Their motor
extracts movement and secretory symbols were also usual
Name of the %Yield and no sign of depression. EEPG+EETG and
Sr. No. Nature Colour
Extract (w/w) g AEPG+AETG even up to the dose level of 2000
Dark mg/kg body weight did not create any behavioural
1. Pet.ether Sticky 2.00
green symptoms or mortality. So 1/10th and 1/5th doses
Dark of (maximum dose tested for each extract) be
2. Chloroform Sticky 6.50
green chosen as medium and high dose and were used in
Dark the current study to investigate nephroprotective
3 Ethanol Sticky 9.50
green and anti- inflammatory actions
Dark
4 Aqueous Sticky 11.00 Anti-inflammatory activity by Carrageenin
brown
induced paw oedema model in rats:
The EEPG+EETG and AEPG+AETG with the
selected doses i.e. 200 and 400 mg/kg consist of
show a important drop in paw oedema volume in

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Carrageenin induced paw oedema in rats at dose have significantly reduced oedema volume
different time interval. Results are tabulated in by 13.9, 24.28, 15.11, 17.29.
Table No. 4.7. Diclofenac sodium (30 mg/kg) was % respectively noted as time reliant result.
used as typical situation and it has considerably During 3rd h of study EEPG+EETG and
reduced paw oedema volume by 17.81% at 1st h, AEPG+AETG with 200mg/kg and 400 mg/kg
27.58% at 2nd h, 50.09% at 3rd h and74.13 % at doses have considerably reduced oedema volume
4th h, which was created to be a moment by 32.07, 34.33, 38.08,
dependent result. 47.46 % correspondingly noted as time dependent
During 1st h of study EEPG+EETG and result.
AEPG+AETG with 200mg/kg and 400 mg/kg During 4th h of study EEPG+EETG and
doses have extensively reduced oedema volume by AEPG+AETG with medium and high doses have
3.43, 10.72, 3.43, 7.72 % respectively noted as significantly reduced oedema volume 51.72,
time dependent result. 56.89, 55.17, 60.34 % respectively which was
During 2nd h of study EEPG+EETG and record as time dependent effect and result be
AEPG+AETG with 200mg/kg and 400 mg/kg graphically represent in Fig No.:4.5.
Table No. 4. Anti-inflammatory effect of EEPG+EETG and AEPG+AETG on paw volume in Carrageenin
induced paw edema in rats
Paw oedema volume
Groups Treatment
60 min 120 min 180 min 240 min
Control Carragenin 0.466±0.042 0.4833±0.047 0.533±0.04 0.58±0.030
Standard Diclofenac 0.383±0.030 0.35±0.02236 0.266±0.033 0.15±0.0223
sodium 6
(30mg/kg)
EEPG+EETG 200mg/kg 0.45±0.0428 0.4166±0.047 0.366±0.033 0.2833±0.03
0
EEPG+EETG 400mg/kg 0.416±0.047 0.366±0.033 0.35±0.022 0.25±0.022
AEPG+AETG 200mg/kg 0.45±0.0428 0.41±0.036 0.33±0.0421 0.26±0.049
AEPG+AETG 400mg/kg 0.43±0.0210 0.4±0.042 0.28±0.030 0.233±0.033

Fig. No. 4 Anti-inflammatory effect of EEPG+EETG and AEPG+AETG on pawvolume in

Carrageenin induced paw oedema in rats

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Table No. 5. Percentage reduction of paw volume in Carrageenin induced paw oedema
% Reduction in paw volume
Groups Treatment
60 min 120 min 180 min 240 min
Control Carragenin - - - -
Standard Diclofenac sodium 17.81 27.58 50.09 74.13
(30mg/kg)
EEPG+EETG 200mg/kg 3.43 13.9 32.07 51.72
EEPG+EETG 400mg/kg 10.72 24.28 34.33 56.89
AEPG+AETG 200mg/kg 3.43 15.11 38.08 55.17
AEPG+AETG 400mg/kg 7.72 17.29 47.46 60.34

Fig. No. 5 Percentage reduction of Paw oedema volume in different groups

Nephroprotective activity by Gentamicin Table No: 6 % of Body weight change in
induced nephrotoxicity in albino rat’s model: gentamicin induced Nephrotoxicity in albino rats.
Gentamicin like other aminoglycoside antibiotics % of body weight
Groups
causes nephrotoxicity by inhibiting protein change
synthesis in renal cells. This alters the body weight Normal control 3.44 ± 0.190
Toxic control 9.926± 0.448***
and elevates the usual levels of blood urea and
EEPG+EETG (200mg/kg) 8.216 ± 0.410***
serum creatinine.
EEPG+EETG (400mg/kg) 5.776 ± 0.463**
The administration of ethanolic and aqueous
AEPG+AETG (200mg/kg) 6.3± 0.44
extracts of combined doses (200mg/kg and 400
AEPG+AETG (400mg/kg) 4.4 ± 0.411
mg/kg) of Punica granatum and Tectona grandis Values are expressed in mean ±SEM where n = 6,
has shown a significant reduction in blood urea Significant at P < 0.05*, 0.01** and 0.001***,
and serum creatinine levels as shown in the tables compared to control group.
below.

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Fig. No: 6 % of Body weight changes in special Fig. No: 8. Serum creatinine level of special groups
groups by Gentamycin inducedNephrotoxicity in gentamicin induced Nephrotoxicitymodel
model. DISCUSSION
Table No.: 7 Blood urea and serum creatinine
In the current study the fruit peel of Punica
levels of different groups by gentamicin induced
granatum and bark of Tectona grandis were chosen
Nephrotoxicity
Serum
for the anti-inflammatory activity. It was
Groups Blood urea previously reported that the individuals of the
creatinine
Normal control 31.03± 5.018 1.023± 0.053 Punica granatum and Tectona grandis have potent
Toxic control 70.591 ± 4.064*** 2.05± 0.081*** anti-inflammatory activity. The preliminary
EEPG+EETG phytochemical research reveals the presence of
57.385 ± 11.724** 1.918± 0.07***
(200mg/kg) alkaloids, flavonoids, saponins, tannins and
EEPG+EETG glycosides in both ethanolic and aqueous extracts.
45.916 ± 11.65* 1.53 ± 0.11***
(400mg/kg) Inflammation is a state which is cause by chemical
AEPG+AETG mediators like prostaglandins and cytokinin’s. It is
51.948 ± 3.673*** 1.668± 0.080***
(200mg/kg) report that the phytochemicals like flavonoids,
AEPG+AETG saponins and tannins decrease the inflammation.
38.046 ± 5.280 1.191 ± 0.053
(400mg/kg)
In the current study the above-mentioned
Values are expressed in mean ±SEM where n = 6,
phytochemicals are current in both the extracts this
Significant at P < 0.05*, 0.01** and 0.001***,
can be recognized for its anti- inflammatory
compared to control group.
movement.
The fruit peel Punica granatum be reported to
contain phenolic compounds like flavonoids,
which possess antioxidant property, The fruit peel
of Punica granatum were previously report for its
nephroprotective movement on diabetes-induced
renal damage. Nephrotoxicity is a regular cause of
human being which is mainly caused due to
oxidative stress. In the current study both the
Fig. No:7. Blood urea level of different groups in extracts containing polyphenolic compounds have
gentamicin induced Nephrotoxicity model antioxidant property, so the nephroprotective
activity of the above-mentioned plants Punica

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granatum and Tectona grandis is appropriate to its acute subacute level in naturally infected
antioxidant property. sheep. Natl J Physiol Pharm Pharmacol
CONCLUSION 2022;12(02):115-120
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and inflammation may be due to creation of 2005; 4(19);1-6.
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ACKNOWLEDGEMENTS Bharat BA; From Ancient Medicine to recent
Authors are very grateful to the staff of Medicine: Ayurvedic concept of Health and
Mansarovar Global University College of Their Role in Inflammation and Cancer;
Pharmacy for their technical assistance. Journal of the Society for Integrative
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International Journal of Pharmacognosy and
HOW TO CITE: Dr. Santhosh Suddagoni*, Dr. kapil
Phytochemical Research; 2012;4(1);12-16. malviya, Dr. Santikari Sesha Phanindra, Dr. M. Venkata
72) Juhi M, Asia Y, Rattan deep S and Aradhana; Ramana, Bodige Divya, Dr. Ganesh Akula,
Nephroprotective and Anti-Inflammatory Activity of
phytochemical investigation and in- vitro Combination of Punica Granatum (Fruit Peel) And
antioxidant potential of leaves of Murraya Tectona Grandis (Bark), Int. J. in Pharm. Sci., 2023, Vol
koenigii; International journal of integrative 1, Issue 6, 86-98.
https://doi.org/10.5281/zenodo.8039301
biology; 2009;7(3);171-174.

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