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© 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 297
Anti-anemic Activity Of Polygonum Glabrum
Gouthami K*, Dr I Veena rani, Dr K Vanitha prakash, Department of pharmacology, SSJ college of pharmacy,
Gandipet, Hyderabad, Telangana.
Abstract
In present research work, anti-anemic effect of methanolic extract (ME) of aerial parts of polygonum glabrum (PG)
was studied in phenylhydrazine (PHZ) induced anemic rats. Fifteen albino wistar rats (3 rats in each group)
weighing 100-150 grams were randomly distributed into: normal control group, anemic control group, standard
reference group administered with vitamin B12 syrup and two test control groups administered with 200mg/kg and
400mg/kg of ME of aerial part of PG respectively twice daily. ME of PG was given orally for one month for test
groups and collect blood samples from all groups. Collected blood samples were estimated for complete blood
count includes RBC, Hb, WBC, PCV, MCH, Platelets, MCV and MCHC. It was observed that, test control group
with 400mg/kg of PG dose improves the Hb percentage in PHZ induced anemic rats very similar to standard
reference group. It concludes the anti-anemic effect of phenolics and flavonoids of PG in PHZ induced anemic rats.
Key words: Anti-anemic, Methanolic, Polygonum glabrum, Phenylhydrazine.
1. Introduction
Anemia may occur at any stage of the life cycle, and it is more chances of occurrence in not only in pregnant
women but also in young children. Iron deficiency anemia (IDA) in 2002 was major contributor of global burden
and affected majorly the health condition of people. Anemia is defined as a decrease in the number of red blood
cells (RBCs) there by leads to a decrease in the content of hemoglobin and or a reduction in both the number of
hemoglobin and RBCs. The condition without enough content of hemoglobin to supply oxygen to the various
tissues, anemia occurs. The cause of anemia is loss of blood heavily occurs by different origins like infection of
parasites like ascaris, hookworms, menstruation, and schistosomiasis may decrease content of hemoglobin (Hb) in
blood. Both Acute and severe infections, like tuberculosis, malaria, cancer, and HIV may decrease content of
hemoglobin (Hb) in blood too. The availability of other deficiencies of micronutrients, like vitamins like folic acid,
riboflavin A, B12, and trace elements such as zinc, iron, and copper might raise the anemia risk.
© 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 298
Figure no.1 Anemia in Pregnant women (Bruno de Benoist et al., 2008)
In developing and undeveloped countries, herbal medicines are going to occupy major role in health care system.
2. Methodology
2.1 Plant description:
Leaves of polygonum glabrum (Genus: Polygonum L, Family: Polygonaceae) used in present research work, it is an
annual herb. Usually, grows highly in marshy lands, Plains from the coast to 1400 m height. It is observed in Java,
South China, Africa, Afghanistan, India, Nepal, Sri Lanka, Pakistan, South Japan, Malaysia, Sylhet and where as in
Chittagong; it is observed in ditches and swamps. In India: Maharashtra, Orissa, Punjab, Rajasthan, Sikkim,
Karnataka, Gujarat, Madhya Pradesh, Uttar Pradesh and West Bengal
Figure no.2 Polygonum glabrum
2.2 Extraction: The aerial part of Polygonum glabrum washed with distilled water and shade dried at room
temperature and then powdered mechanically. The Methanolic extract (ME) of aerial part was prepared by
suspending the powder in Methanol by Soxhalation method.
2.3 Phytochemical screening: Phytochemical investigation of Methanolic aerial part extract will be done by
performing various tests such as alkaloids, flavonoids, glycosides, proteins, amino acids, carbohydrates, tannins,
fats, saponins and steroids.
2.4 Animals: Albino wistar rats weighing 100-150 grams were chosen for present research work. Animals were
fasted for food prior to dosing but not water overnight withheld. Three animals were kept for each and every dose.
Animals were observed for behavioral alterations for 4 hours and incase of mortality for 48 hours (O.E.C.D
© 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 299
GUIDELINES 423). All the animals testing were done as per the approval of Institutional Animal Ethical
Committee.
2.5 Anti-anemic studies: Induction of Anemia was done in rats by oral treatment of PHZ at 10mg/kg for eight
days. Rats with hemoglobin (Hb) lesser than 13 g/dl were selected for further studies.
In-vivo Study: Rats were divided into five groups of three rats each and study design was used in present research
work explained in table no.1.
Statistical analysis: Experimental data were analyzed by applying one way analysis of variance (ANOVA) and LSD
multiple range tests to determine significant differences between means.
Table no.1 Study design for anti-anemic activity
GROUPS TREATMENT
Group I Animals served as the positive control (normal animals) and were administered 0.9 % normal saline
solution.
Group II Animals served as the negative control (anemic animals) and were administered 0.9 % normal saline
solution.
Group III Animals treated with Standard Vitamin B12 Syrup
Group IV Animals were administered graded doses of 200 mg/kg body weight of the Methanolic aerial part
extract of Polygonum glabrum respectively twice daily (morning and evening).
Group V Animals were administered graded doses of 400 mg/kg body weight of the Methanolic aerial part
extract of Polygonum glabrum respectively twice daily (morning and evening).
Analysis of Hematological Parameters:
1. Count of The red blood cell (i.e. RBC)
2. Count of White blood cell (i.e. WBC)
3. Hemoglobin concentration (i.e. Hb)
4. Haematocrit were estimated at end of study by Semi Auto-Biochemistry Analyzer.
3. Results
3.1 Percentage Yield: The extract showed maximum of 14 % yield.
© 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 300
3.2 Phytochemical screening:
Table No.2 Phytochemical Analysis
TEST RESULT
ALKALOIDAL TEST Positive
CARBOHYDRATES TEST Positive
STEROIDS TEST Negative
GLYCOSIDES TEST Positive
SAPONINS TEST Positive
FLAVONOIDS TEST Negative
TRITERPINOIDAL TEST Negative
TANNINS TEST Positive
PROTIEN& AMINOACIDS TEST Positive
3.3 Pharmacological investigations
3.3.1 Acute toxic study: Acute toxic study was done by following OECD guidelines. The Lethal dose LD50 cut- off
doses obtained for various extracts are enlisted in Table no.3.
Table No. 3 Acute oral toxic study of various extracts of Polygonum glabrum
Extract GHS classification LD50 cut-off dose Therapeutic dose
ME leaves Class 4 2000 mg/kg b.w 200 mg/kg b.w
ME leaves Class 4 2000 mg/kg b..w 400 mg/kg b.w
*1/5th
and 1/10th
of this LD50 dose were taken as therapeutic dose for subsequent pharmacological
screenings
GHS- Globally harmonized system
© 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 301
3.3.2 Effect of extract on body weight parameters of PHZ induced anemic rats:
Figure no.3 Effect of extract on body weight parameters of rat
(Values are Mean ± SEM (n = 6), analyzed by one way ANOVA followed Dunnett’s test Significance:
*** P<0.001, ** P<0.01, * P<0.05 from control)
3.3.3 Effect of extract on hematological parameters of PHZ induced anemic rats:
© 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 302
Figure no.4 Effect of extract on PHZ induced anemic rats
(Values are Mean ± SEM (n = 6), analyzed by one way ANOVA followed Dunnett’s test
Significance: *** P<0.001, ** P<0.01, * P<0.05 from control)
4. Summary and conclusion
Phenylhydrazine generates both hydroxyl and aryl radicals after incubation with rat liver microsomes.
Hemolytic injury induced by PHZ considered to be originated from oxidative changes to proteins of red blood cell
compared to lipids of membrane. In this current investigation, PHZ changed the parameters of hematology due to
hemolysis effects featured by reduction in hemoglobin i.e. Hb amount, counts of all RBC.
The enhancement in the indices of hematology exhibited by extract could be associated with the phenolics,
minerals constituents of the aerial parts of polygonum glabrum. The phenolics and flavonoids have showed anti
anemic effect. These compounds could have direct impact on the hemolysis protection through species of reactive
oxygen produced by PHZ.
5. References
Rang, H.P, Dale, M.M., (2003), Pharmacology, (6), USA, Churchil Livingstone Elsevier.
Williams, D. A, Lemake, T. L., (2002), Foye’s Principles of Medicinal Chemistry, (5), New York, Lippincott
Williams and Wilkins.
Watkins, W.J., Renau, T.E., (2003), Burger’s Medicinal Chemistry and Drug discover, (6),New York, A Wiley-
Interscience Publication.
Seth, S. D, Seth, V., (2009), Textbook of Pharmacology, (3), Noida, India, a division of Reed Elsevier India Private
Limited.
Chauhan, S.P., Sheth, N.R., Suhagia B.N., (2015), Hematinic effect of fruits of opuntia elatior mill on
phenylhydrazine induced anemia in rats, Ayujournal, 36(2), 208-13, doi: 10.4103/0974-8520.175549.
Raghavendra, H. L., Yogesh, H. S., Gopalakrishna, B., Chandrashekar, V. M., Sathishkumar, B. P., Vadlapudi
Kumar, (2009), An overview of herbal medicine, International Journal of Pharmaceutical Sciences,1(1), 1 - 20.
Newman, D.J., Cragg, G.M., (2007), Natural products as sources of new drugs over the last 25 years, Journal of
Natural Product, 70, 461–477, https://doi.org/10.1021/np068054v.
Vaidya, A.D.B., Devasagayam, T.P.A., (2007), Current status of herbal drugs in India: an overview, Journal of
Clinical Biochemistry and Nutrition, 41, 1–11, https://doi.org/10.3164/jcbn.2007001.
Silja, V.P., Verma, S.K., Mohanan, K.V., (2008), Ethanomedicinal plant knowledge of the Mullu kuruma tribe of
Wayanad district, Kerala, Indian Journal of Traditional Knowledge, 7(4), 604-612,
http://hdl.handle.net/123456789/2389.

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Anti-anemic Effect of Polygonum glabrum in Rats

  • 1. © 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138) IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 297 Anti-anemic Activity Of Polygonum Glabrum Gouthami K*, Dr I Veena rani, Dr K Vanitha prakash, Department of pharmacology, SSJ college of pharmacy, Gandipet, Hyderabad, Telangana. Abstract In present research work, anti-anemic effect of methanolic extract (ME) of aerial parts of polygonum glabrum (PG) was studied in phenylhydrazine (PHZ) induced anemic rats. Fifteen albino wistar rats (3 rats in each group) weighing 100-150 grams were randomly distributed into: normal control group, anemic control group, standard reference group administered with vitamin B12 syrup and two test control groups administered with 200mg/kg and 400mg/kg of ME of aerial part of PG respectively twice daily. ME of PG was given orally for one month for test groups and collect blood samples from all groups. Collected blood samples were estimated for complete blood count includes RBC, Hb, WBC, PCV, MCH, Platelets, MCV and MCHC. It was observed that, test control group with 400mg/kg of PG dose improves the Hb percentage in PHZ induced anemic rats very similar to standard reference group. It concludes the anti-anemic effect of phenolics and flavonoids of PG in PHZ induced anemic rats. Key words: Anti-anemic, Methanolic, Polygonum glabrum, Phenylhydrazine. 1. Introduction Anemia may occur at any stage of the life cycle, and it is more chances of occurrence in not only in pregnant women but also in young children. Iron deficiency anemia (IDA) in 2002 was major contributor of global burden and affected majorly the health condition of people. Anemia is defined as a decrease in the number of red blood cells (RBCs) there by leads to a decrease in the content of hemoglobin and or a reduction in both the number of hemoglobin and RBCs. The condition without enough content of hemoglobin to supply oxygen to the various tissues, anemia occurs. The cause of anemia is loss of blood heavily occurs by different origins like infection of parasites like ascaris, hookworms, menstruation, and schistosomiasis may decrease content of hemoglobin (Hb) in blood. Both Acute and severe infections, like tuberculosis, malaria, cancer, and HIV may decrease content of hemoglobin (Hb) in blood too. The availability of other deficiencies of micronutrients, like vitamins like folic acid, riboflavin A, B12, and trace elements such as zinc, iron, and copper might raise the anemia risk.
  • 2. © 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138) IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 298 Figure no.1 Anemia in Pregnant women (Bruno de Benoist et al., 2008) In developing and undeveloped countries, herbal medicines are going to occupy major role in health care system. 2. Methodology 2.1 Plant description: Leaves of polygonum glabrum (Genus: Polygonum L, Family: Polygonaceae) used in present research work, it is an annual herb. Usually, grows highly in marshy lands, Plains from the coast to 1400 m height. It is observed in Java, South China, Africa, Afghanistan, India, Nepal, Sri Lanka, Pakistan, South Japan, Malaysia, Sylhet and where as in Chittagong; it is observed in ditches and swamps. In India: Maharashtra, Orissa, Punjab, Rajasthan, Sikkim, Karnataka, Gujarat, Madhya Pradesh, Uttar Pradesh and West Bengal Figure no.2 Polygonum glabrum 2.2 Extraction: The aerial part of Polygonum glabrum washed with distilled water and shade dried at room temperature and then powdered mechanically. The Methanolic extract (ME) of aerial part was prepared by suspending the powder in Methanol by Soxhalation method. 2.3 Phytochemical screening: Phytochemical investigation of Methanolic aerial part extract will be done by performing various tests such as alkaloids, flavonoids, glycosides, proteins, amino acids, carbohydrates, tannins, fats, saponins and steroids. 2.4 Animals: Albino wistar rats weighing 100-150 grams were chosen for present research work. Animals were fasted for food prior to dosing but not water overnight withheld. Three animals were kept for each and every dose. Animals were observed for behavioral alterations for 4 hours and incase of mortality for 48 hours (O.E.C.D
  • 3. © 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138) IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 299 GUIDELINES 423). All the animals testing were done as per the approval of Institutional Animal Ethical Committee. 2.5 Anti-anemic studies: Induction of Anemia was done in rats by oral treatment of PHZ at 10mg/kg for eight days. Rats with hemoglobin (Hb) lesser than 13 g/dl were selected for further studies. In-vivo Study: Rats were divided into five groups of three rats each and study design was used in present research work explained in table no.1. Statistical analysis: Experimental data were analyzed by applying one way analysis of variance (ANOVA) and LSD multiple range tests to determine significant differences between means. Table no.1 Study design for anti-anemic activity GROUPS TREATMENT Group I Animals served as the positive control (normal animals) and were administered 0.9 % normal saline solution. Group II Animals served as the negative control (anemic animals) and were administered 0.9 % normal saline solution. Group III Animals treated with Standard Vitamin B12 Syrup Group IV Animals were administered graded doses of 200 mg/kg body weight of the Methanolic aerial part extract of Polygonum glabrum respectively twice daily (morning and evening). Group V Animals were administered graded doses of 400 mg/kg body weight of the Methanolic aerial part extract of Polygonum glabrum respectively twice daily (morning and evening). Analysis of Hematological Parameters: 1. Count of The red blood cell (i.e. RBC) 2. Count of White blood cell (i.e. WBC) 3. Hemoglobin concentration (i.e. Hb) 4. Haematocrit were estimated at end of study by Semi Auto-Biochemistry Analyzer. 3. Results 3.1 Percentage Yield: The extract showed maximum of 14 % yield.
  • 4. © 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138) IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 300 3.2 Phytochemical screening: Table No.2 Phytochemical Analysis TEST RESULT ALKALOIDAL TEST Positive CARBOHYDRATES TEST Positive STEROIDS TEST Negative GLYCOSIDES TEST Positive SAPONINS TEST Positive FLAVONOIDS TEST Negative TRITERPINOIDAL TEST Negative TANNINS TEST Positive PROTIEN& AMINOACIDS TEST Positive 3.3 Pharmacological investigations 3.3.1 Acute toxic study: Acute toxic study was done by following OECD guidelines. The Lethal dose LD50 cut- off doses obtained for various extracts are enlisted in Table no.3. Table No. 3 Acute oral toxic study of various extracts of Polygonum glabrum Extract GHS classification LD50 cut-off dose Therapeutic dose ME leaves Class 4 2000 mg/kg b.w 200 mg/kg b.w ME leaves Class 4 2000 mg/kg b..w 400 mg/kg b.w *1/5th and 1/10th of this LD50 dose were taken as therapeutic dose for subsequent pharmacological screenings GHS- Globally harmonized system
  • 5. © 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138) IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 301 3.3.2 Effect of extract on body weight parameters of PHZ induced anemic rats: Figure no.3 Effect of extract on body weight parameters of rat (Values are Mean ± SEM (n = 6), analyzed by one way ANOVA followed Dunnett’s test Significance: *** P<0.001, ** P<0.01, * P<0.05 from control) 3.3.3 Effect of extract on hematological parameters of PHZ induced anemic rats:
  • 6. © 2019 IJRAR June 2019, Volume 6, Issue 2 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138) IJRAR19K6346 International Journal of Research and Analytical Reviews (IJRAR)www.ijrar.org 302 Figure no.4 Effect of extract on PHZ induced anemic rats (Values are Mean ± SEM (n = 6), analyzed by one way ANOVA followed Dunnett’s test Significance: *** P<0.001, ** P<0.01, * P<0.05 from control) 4. Summary and conclusion Phenylhydrazine generates both hydroxyl and aryl radicals after incubation with rat liver microsomes. Hemolytic injury induced by PHZ considered to be originated from oxidative changes to proteins of red blood cell compared to lipids of membrane. In this current investigation, PHZ changed the parameters of hematology due to hemolysis effects featured by reduction in hemoglobin i.e. Hb amount, counts of all RBC. The enhancement in the indices of hematology exhibited by extract could be associated with the phenolics, minerals constituents of the aerial parts of polygonum glabrum. The phenolics and flavonoids have showed anti anemic effect. These compounds could have direct impact on the hemolysis protection through species of reactive oxygen produced by PHZ. 5. References Rang, H.P, Dale, M.M., (2003), Pharmacology, (6), USA, Churchil Livingstone Elsevier. Williams, D. A, Lemake, T. L., (2002), Foye’s Principles of Medicinal Chemistry, (5), New York, Lippincott Williams and Wilkins. Watkins, W.J., Renau, T.E., (2003), Burger’s Medicinal Chemistry and Drug discover, (6),New York, A Wiley- Interscience Publication. Seth, S. D, Seth, V., (2009), Textbook of Pharmacology, (3), Noida, India, a division of Reed Elsevier India Private Limited. Chauhan, S.P., Sheth, N.R., Suhagia B.N., (2015), Hematinic effect of fruits of opuntia elatior mill on phenylhydrazine induced anemia in rats, Ayujournal, 36(2), 208-13, doi: 10.4103/0974-8520.175549. Raghavendra, H. L., Yogesh, H. S., Gopalakrishna, B., Chandrashekar, V. M., Sathishkumar, B. P., Vadlapudi Kumar, (2009), An overview of herbal medicine, International Journal of Pharmaceutical Sciences,1(1), 1 - 20. Newman, D.J., Cragg, G.M., (2007), Natural products as sources of new drugs over the last 25 years, Journal of Natural Product, 70, 461–477, https://doi.org/10.1021/np068054v. Vaidya, A.D.B., Devasagayam, T.P.A., (2007), Current status of herbal drugs in India: an overview, Journal of Clinical Biochemistry and Nutrition, 41, 1–11, https://doi.org/10.3164/jcbn.2007001. Silja, V.P., Verma, S.K., Mohanan, K.V., (2008), Ethanomedicinal plant knowledge of the Mullu kuruma tribe of Wayanad district, Kerala, Indian Journal of Traditional Knowledge, 7(4), 604-612, http://hdl.handle.net/123456789/2389.