Immunomodulatory activity of Anacardium occidentale L. in Laboratory animals

 

Salma A. Bagwan*, Nilofar S. Naikwade

Department of Pharmacology, Appasaheb Birnale College of Pharmacy, Sangli.

*Corresponding Author E-mail: salmabawan233@gmail.com

 

ABSTRACT:

Aim: To study the phytochemical analysis and immunomodulatory activity of ethyl acetate extract of nut of Anacardium occidentale Linn. Methods: A laboratory based experimental method was used to evaluate immunomodulatory activity by dividing animals into six different groups, comprising six animals in each group. The immunomodulatory activities were determined by using carbon clearance test, delayed type hypersensitivity (DTH) test and In vivo antibody (HA) titer response to SRBCs. Result: The response produced by oral administration of ethyl acetate extract of nut of A.occidentale showed a significant dose dependent effect by increase in Phagocytic activity, DTH response and augmentation of humoral immune response to sheep red blood cells. Conclusion: Ethyl acetate extract of nut of A. occidentale showed a significant immunostimulatory effect on both the humoral and cell-mediated immune systems in the Wistar albino rats.

 

KEYWORDS: Anacardium occidentale L., Immunomodulatory activity, Phytochemical analysis, Levamisole, Cyclophosphamide.

 

 


1. INTRODUCTION: 

Recently, a healthy diet paradigm has been very important for maintaining normal healthy wellbeing, which places more emphasis on the positive aspects of diet. Change in lifestyle and change in nutritional habits may have a positive or negative impact on health. Nutritional status of the body also determines immunological defense against certain pathogenic diseases.1 In the 21st century the world is facing the incidence of a large number of infectious diseases. In 2019, coronavirus diseases (SARS-COV-2) is a pandemic which causes severe acute respiratory syndrome. About 15-18 million people are infected and many of them have died. A variety of antibiotic and antiviral drugs are available on the market which are effective at resisting infection, but after prolonged use they become ineffective due to thedevelopment of antibiotic resistance to the pathogen. Not any treatment has been definitively shown effective.

 

 

However, the prevention is better than cure strategies such as vaccination and the use of immunomodultors might have beneficial effects on controlling the pathological immune response. So, recently researchers might have been thinking more about the method of immunomodultors as a future treatment.2 Abnormalities of the immune system that result in defective immune responses make individuals susceptible to infections by viruses, bacteria, fungi and parasites.3

 

In immunopharmacology we study the effect of the different drugs which modify immune mechanisms in the host body. The world health organization (WHO) estimates that about 4 billion people, of the world population, presently use herbal medicine for primary health care.4 The certain herbs which possess a wide variety of therapeutic properties which include some nutraceuticals. Nutraceuticals are pharmaceutically blended products which possess both nutritional as well as medicinal properties, so that such a product is designed to improve the overall physical health, to fight against day-to-day challenges such as stress and also increase durability of survival. Some herbs show a wide variety of therapeutic values, such as immunity booster, anti-diabetic, anticancer, antimicrobial, and gastro-protective action. Herbs could be the option/choice to formulate as Nutraceuticals.

Nowadays, the problem faced by people due to a weaker immune system is that individuals make them more prone to frequent infection.5 One of the main disadvantages of the use of anti-biotic is that it can develop microbial resistance when used frequently. Like an army of soldiers, this system has many vital functions and uses the body’s defense to guard against damage, disease and infectious organisms. These immune cells are activated by the presence of cytokines, such as Interferon-gamma (IFN-γ) and Tumor necrosis factor-α (TNF-α).6,7 so it is important to increase the overall natural resistance of individuals to fighting against infectious agents. The purpose of the study is to find out the immunopharmacological activity of Anacardium occidentale as a nutraceuticals which has been suggested in the Indian system of medicine for curing a number of diseases.

 

2. MATERIALS AND METHODS:

2.1 Collection of plant:

The nuts of A. occidentale were collected from the region of Ratnagri, Maharashtra. After collection, the plants were authenticated by the botanical Service of India Pune, Maharashtra (BSI/WRC/IDEN.CER./2020/95 dated 01/10/2020).

 

2.2 Sample preparation:

The nuts of A. occidentale was thoroughly clean. They are cut into small pieces, grinded into powder and stored in an airtight container.

 

2.3 Extraction of sample:

The dried powder samples of nuts of A. occidentale were undergone soxhlet extraction using ethyl acetate solvent. The concentrated extract was collected and stored in an airtight container for further use.8,9

 

2.4 Preliminary phytochemical investigation:

Phytochemical investigations were performed to identify the presence of various chemical constituents in the extracts, as per Khandelwal 2012.10

 

2.5 Animals used:

Wistar albino rats (weighing approximately 150 to 200g) were taken from the animal house of Appasaheb Birnale College of Pharmacy, Sangli, Maharashtra. The experimental protocol was approved by the institutional animal ethics committee (IAEC/ABCP/16/2020-21) and the care of laboratory animals was taken as per the guidelines of CPCSEA.

 

2.6 Acute toxicity studies:

Ethyl acetate extract of nut of A. occidentale was tested for acute toxicity studies as per procedure given in OECD guidelines 425. Doses selected 150, 300 and 450 mg/kg body weight of animals and were observed for any sign of toxicity and mortality.11,12

 

2.7 Antigenic material:

2.7.1 Preparation of sheep RBCs:

Sheep blood was collected in sterile Alsever’s solution in 1:1 proportion, Alsever’s solution (freshly prepared) blood was kept in the refrigerator and processed for the preparation of SRBCs batch, by centifugating at 2000 rpm for 10min and washing with physiological saline 4-5 times and then suspending into buffered saline for further use13

 

2.7.2 Preparation of carbon ink suspension:

Camlin ink was diluted eight times with saline and used for carbon clearance test in a dose of 10μl/gm. body weight of rat.13

 

2.8 Grouping and treatment schedule:

Table 1: Grouping and treatment schedule

Groups

Test substance

Dose (P.O)

Group I

Control

10 ml/kg

Group II

Levamisole (Std-1)

50 mg/kg

Group III

Cyclophosphamide (Std-2)

50 mg/kg

Group IV

EAAO-150

150 mg/kg

Group V

EAAO-300

300 mg/kg

Group VI

EAAO-450

450 mg/kg

 

2.9 In vivo immunopharmacological activity:

1.9.1 Carbon clearance test

1.9.2 Delayed type hypersensitivity test

1.9.3 In vivo antibody (HA) titer response to SRBCs

 

2.9.1 Carbon clearance test.

Phagocytic activity of reticuloendothelial system was assay by carbon clearance test; phagocytic index is used to calculate as a rate of carbon elimination by reticuloendothelial system.

 

Procedure

1.     In this test Animals were divided into six groups comprising 6 animals in each. As per grouping schedule mention in table 1

2.     Carbon ink suspension was injected via tail vein to each rat 48 hours, after the seven days of treatment.

3.      Blood sample (25 μl) were then withdrawn from the retro-orbital plexus under mild ether anesthesia at 5 and 15 min after injection of colloidal carbon ink lysed in 0.1% sodium carbonate solution (3 ml).

4.     The optical density was measured spectrophotometrically at 660 nm.

5.     The phagocytic activity was calculated using the following formula14,15,16,17

 

K= Log OD1-Log OD2/ t2 - t1

Where ODI and OD2 are the optical densities at time t1 and t2, respectively

2.9.2. Delayed type hypersensitivity test:

Delayed hypersensitivity reaction is a reaction of cell mediated immunity and become visible only after 16-24 hrs.

 

Procedure:

1.     In this test Animals were divided into six different groups each comprising 6 animals. As per grouping schedule mention in table 1.

2.     Immunized Rat with 0.1ml of 20% SRBCs in normal saline intraperitonially on 14th day of the study.

3.     On day 21st, animals from all groups get challenge with 0.03ml of 1% SRBCs in sub plantar region of right hind paw. Foot pad reaction was assessed after 4h, 8h and 24 h i.e., on 22nd day. Increase in footpad edema was measured with the help of vernier        caliper. 14,15,16,17

 

2.9.3.         In vivo antibody (HA) titer response to SRBCs:

The highest dilution of a sample at which clumping is seen is considered as haemagglutination.

 

Procedure:

1.     The animals were immunized by injecting 0.1 ml of SRBC suspension containing 0.5 × 109 cells intra-peritoneally on day 0.

2.     Blood samples were collected in micro centrifuge tubes from individual animal by retro-orbital puncture on day 11.

3.     The blood samples were centrifuged and serum was obtained.

4.     Antibody levels were determined by the haemagglutination technique. Equal volumes of individual serum samples of each group were pooled. Two-fold serial dilutions of pooled serum samples made in 25 μl volume of normal saline in micro-titration plates were added to 25 μl of 1% suspension of SRBCs in saline.

5.     After mixing, the plates were incubated at 37°C for 1 h and examined for haemagglutination under a microscope.

6.     The reciprocal of the highest dilution of the test serum agglutination was taken as the antibody titre.18,19

 

2.9 Statistical analysis:

Mean Value ±SEM was used to express the test outcome. The one way analysis of variance (ANOVA) technique was used to estimate the variation in a collection of data. The individual comparison of group mean value were done by Dunnet’s Test. Statistics were considered significant when the P< 0.05.

 

 

3. RESULTS:

Table 2: Phytochemical investigation of ethyl acetate extract of nut of A. occidentale

S. No

Test

Result

1

Test for carbohydrate

Molish’s test ( General test)

Benedict’s test

 

-

-

2

Test for protein and amino acid

Biuret test ( General test)

Millon’s test

Ninhydrin test

 

-

-

-

3

Test for glycoside

For cardiac glycoside

Baljets test

Legal test

Keller-Killiani test

For anthraquinone glycoside

Borntargers test

Modified Borntargers

For Saponin glycoside

Foam test

 

 

++

-

-

 

++

++

 

-

4

Test for flavonoid

Sulphuric acid test

Shinoda test

Lead acetate solution test

 

+++

+++

-

5

Test for alkaloid

Dragendroff’s test

Mayer’s test

Hager’s test

Wagnar’s test

 

-

-

-

-

6

Test for tannin and Phenolic compound

5% Fecl3

Lead acetate solution

Bromine water

Nitric acid

 

+++

+++

+++

-

7

Test for steroids

Salkowaski test

Libermannburchard test

 

-

-

8

Test for Vitamin C

+

Note: (-): Absent, (+): Present, (++): Moderate Present, (+++): Strongly Present

 

The phytochemical screening of ethyl acetate extract of  A. occidentale were done by qualitative chemical test and results obtained were showed in table. [Table 2]

 

1.     Carbon clearance test:

Table 3 Result of carbon clearance test

Groups

Treatments

Dose and route of administration (P.O)

Phagocytic index

(Mean±SEM)

I

Control

10ml/kg

0.0122± 0.0014

II

Levamisole

(Std-1)

50mg/kg

0.0729±0.0038****

III

Cyclophosphamide (Std-2)

50mg/kg

0.0067±0.00005ns

IV

EAAO

150mg/kg

0.0441±0.0015****

V

EAAO

300mg/kg

0.0529±0.0025****

VI

EAAO

450mg/kg

0.0603±0.00308****

Value are expressed as (Mean ± SEM)n=6 ****p<0.0001 statistically significant when compared with control group by ANOVA followed by Dennett test.

 

 


 

Fig 1: Graphical representation of carbon clearance test.

 

2) Delayed type hypersensitivity test:

Table 4: Result of delayed type hypersensitivity test

Groups

Treatments

Dose and route of administration (P.O.)

% Increment in foot pad thickness

4 h

8 h

24 h

I

Control

10ml/kg

7.20±0.5510

14.67±0.4261

20.39±0.4597

II

Levamisole (Std-1)

50mg/kg

26.875± 0.57698****

34.94± 0.4822****

44.50± 0.4191****

III

Cyclophosphamide (Std-2)

50mg/kg

5.83±0.4890ns

9.382±0.5684****

11.01±0.7774****

IV

EAAO

150mg/kg

11.76±0.0645****

21.16±0.1377****

23.63±0.1476****

V

EAAO

300mg/kg

15.03±0.1527****

24.86±0.2016****

28.06±0.0985****

VI

EAAO

450mg/kg

18.90±0.1337****

28.30±0.1291****

32.41±0.1545****

Values are expressed as (Mean ± SEM) n=6 ****p<0.0001 statistically significant when compared with control group by ANOVA followed by Dunnett test.


 

 

Fig: 2 Graphical representation of delayed type hypersensitivity test at 4 h.

 

Fig: 3 Graphical representation of delayed type hypersensitivity test at 8 h.

 

 

Fig: 4 Graphical representation of delayed type hypersensitivity test at 24 h.

 

3) In vivo antibody (HA) titer response to SRBCs.

Table 5 Result of In vivo antibody (HA) titer response to SRBCs

Groups

Treatments

Dose and route of administration (P.O.)

HA Titer

(Mean ± SEM)

I

Control

10 ml/kg

9.33±1.3385

II

Levamisole

(Std-1)

50 mg/kg

469.33 ± 42.8326****

III

Cyclophosphamide (Std-2)

50 mg/kg

5.33±0.8465ns

IV

EAAO

150 mg/kg

58.66±5.3540ns

V

EAAO

300 mg/kg

117.33± 10.7081**

VI

EAAO

450 mg/kg

213.33± 27.0897****

Values are expressed as (Mean±SEM) n=6 ****p<0.0001 statistically significant when compared with control group by ANOVA followed by Dunnett test.

 

 

Fig: 5 Graphical representation of In vivo antibody (HA) titer response to SRBCs

 

4.    DISCUSSION:

The various plants, fruits, vegetables, nutraceuticals used in the Indian medical system of Ayurveda exhibit a wide range of pharmacological qualities19. Finding plant phytochemicals with immunomodulatory action that might one day be used as alternative medications is gaining more and more attention in the modern era. The scientific findings from the various studies of the nutsof  A. occidentale plant is widely grown for therapeutic purposes20. The major goal of the present study was to concentrate on the immunomodulatory activity of ethyl acetate extract of the nut of A. occidentale with particular attention to its potential immunomodulatory and protective effects in animal models. Acute oral toxicity studies of ethyl acetate extract of A. occidentale were performed by OECD guideline 425. We had selected doses 150, 300 and 450 mg/kg which were safe and the dose did not show any signs of toxicity or mortality.20,21 The study was conducted using five different methods, each of which offers details on the impact on various immune system components. The results of the study suggested that the selected dose 450 mg/kg of the ethyl acetate extract of A. occidentale stimulated overall humoral and cell mediated immune response due to presence of different phytoconstituents like vitamin A vitamin C, fat, protein, carbohydrate, calcium, phosphorus, iron tannins, cardol, anacardic acid, triglycerides, fatty acids, alkyl-substituted phenol, water, cholesterol and the main constituents of the free fatty acids are palmitic and oleic acids. A. occidentale also reported different pharmacological properties like anti-microbial, anti-bacterial, anti-septic, anti-oxidant, anticancer, anti-diabetic and anti-ulcerative property.

 

4.1 Phytochemical investigation:

Phytochemical studies showed ethyl acetate extract of nuts of A. occidentale showed presence of flavonoids, Vitamins C, polyphenols, Glycoside and terpenoids, which may  responsible to modulate immune response.

 

4.2 Carbon clearance test:

Reticuloendothelial system (RES) consist of mononucleated phagocytes and which are mainly responsible to causes phagocytosis. Exogenously administered antigen, i.e. colloidal carbon ink preparation, are recognized as foreign invaders and served by RES through this process of phagocytosis. An increase in removal of carbon particle from the blood stream indicated increase in phagocytic activity.22,23 Effect of EAAO on the phagocytic activity by carbon clearance test is shown in (Table 3). In carbon clearance test EAAO treated all groups exhibited significantly high phagocytic index (p<0.0001) when compare with control group. EAAO treated group showed phagocytic index 0.0441±0.0015**** of dose 150mg/kg, 0.0529±0.0025**** of dose 300mg/kg and 0.0603±0.00308**** of dose 450mg/kg which indicated stimulation of reticuloendothelial system to when compare with control. Similarly Cyclophosphamide treated group showed phagocytic index 0.0067±0.00005ns which showed not significant effect on reticuloendothelial system when compare with control group. Increases phagocytic index of ethyl acetate extract of A. occidentale of dose 450mg/kg close to results obtained by standard levamisole which is the indicator of increased in vivo phagocytic activity and granulopoetic system competency in removing foreign particles, which is a sign ofincreased immune response to foreign particles or antigens.

 

4.3 Delayed type hypersensitivity test:

Delayed hypersensitivity test (DTH) is a type IV hypersensitivity reaction which was used to evaluate the skin hypersensitivity reaction after inoculation of the foreign antigen.24 This antigen triggers immune response specifically memory T cells that in responsible for margination and inflammation at a site of antigen challenge (SRBCs) that more prominently observed after 24hrs. Further, DTH response also helps to improve lytic enzyme concentration as well as phagocytic activity for more efficient destruction of microbes. Effects of EAAO on cell mediated immune response by DTH induces footpad edema is shown in (Table 4)  EAAO treated groups significantly showed an increase in footpad edema (p<0.0001) potentiating delayed type hypersensitivity response when compare with control group at the interval of 4h, 8h and 24h which indicates the activation of cellular immune response, release of cytokine increases permeability to vessels and leads to vasodilation, increase in accumulation of phagocyte leading  to inflammation. The EAAO treated groups showed % increment in footpad edema 11.76±0.0645****, 15.03±0.1527**** and 18.90±0.1337**** of dose 150, 300, 450 mg/kg at 4 h respectively. After 8 h EAAO treated group showed % increment in foot pad thickness 21.16±0.1377****, 24.86±0.2016**** and 28.30±0.1291**** of dose 150, 300, 450 mg/kg respectively. DTH response prominently showed after 24h which showed significant repose on foot pad thickness. The EAAO treated groups showed % increment in foot pad thickness 23.63±0.1476****, 28.06±0.00985**** and 32.41±0.1545**** at a doses 150, 300, 450mg/kg respectively when compare with control. Increases DTH response of ethyl acetate extract of .A occidentale of dose 450mg/kg close to results obtained by standard Levamisole at 24h which is the indicator of increased in cell mediate immune response to foreign particles or antigens.

 

4.4 In vivo antibody (HA) titer response to SRBCs:

HA titer response is used to assays humoral immune response, it involve interaction of B-cell with the antigen and their subsequent proliferation ,differentiation into plasma cell which secrete antibodies[25,26]The augmentation of the humoral immune response to SRBCs  indicated the enhanced responsiveness of T and B lymphocyte ,increase in antibody titer. Humoral antibody response to SRBCs challenge was found to be significantly (p<0.0001) increase by EAAO treated groups when compare with control (Table 5) EAAO 450 mg/kg showed HA titer response 213.33±27.0897**** and EAAO 300mg/kg showed 117.33±10.7081** where EAAO 150mg/kg showed not significant effect over haemagglutination response when compare with control.

 

Overall immunostimulant effect were observed by EAAO treated groups of different doses positive response were observed over reticuloendothelial system, delayed type hypersensitivity reaction andincrease in antibody response against antigen when compare with control as well as Levamisole (Std-1) it indicate ethyl acetate extract of nuts of A. occidentale showed immunomodulatory activity.

 

5.    CONCLUSION:

Nutraceuticals play a very important role in day to day life for maintaining normal healthy body function. They also have an important role in boosting the immune response of our body to defense against pathogenic infection. According to the academy of nutrition and dietetics, good nutrition habits such as eating a healthy and balanced diet are crucial to promoting a strong immune system, and   suggested essential nutrients that support immune health include vitamins A, B6, C, and E, as well as folic acid, zinc, selenium, and iron A. occidentale is nutraceuticals which possesses number of beneficial effect on human body it shows anti-microbial, anti-oxidant, anticancer, anti-diabetic and anti-ulcerative property. Immunomodulatory potential of A. occidentale could be attributed to the presence of flavonoids, Vitamins, polyphenols, Glycoside and terpenoids, which may modulate and potentiate humoral as well as cellular immunity. This emphasizes the future scope of this study.

 

6.    ACKNOWLEDGMENTS:

The authors would like to thank Appasaheb Birnale College of Pharmacy, Sangli for their kind support during studies.

 

7. REFERENCE:

1.      Das L, Bhaumik E, Raychaudhuri U, Chakraborty R. Role of nutraceuticals in human health. J Food Sci Technol. 2012; 49(2): 173-83

2.      https://www.who.int>docs coronavirus diseases (covid -19) weekly report ( dated as 8/8/22)

3.      Gayathiri. K, Gopi Sudheer Kumar. J, Kavimani. S. Evaluation of Immunomodulatory activity of Salvia hispanica. L Seeds. Research J. Pharm. and Tech. 2017; 10(12): 4255-60. doi: 10.5958/0974-360X.2017.00779.X

4.      Syed Safiullah Ghori, Ayesha Ruqsar, Mohammed Akram, Zehra Fatima,Mir Iftekhar Ali Arafath. Evaluation of Immunomodulatory activity of Ficus dalhousiae Miq leaves Methanolic extract against Cyclophosphamide Induced Myelosuppression. Research J. Pharm. and Tech. 2018; 11(8): 3427-30. doi: 10.5958/0974-360X.2018.00632.7

5.      Mykola А. Komisarenko, Ivan M. Polischuk, Taras V. Upyr, Narzullos B. Saidov. Study of Amino Acid Composition and Immunomodulatory Activity of Rubus idaeus Alcoholic Extract. Research J. Pharm. and Tech. 2021; 14(3): 1329-32. doi: 10.5958/0974-360X.2021.00236.5

6.      Shantilal Singune, Jai S Vaghela. Investigation of Immunomodulatory Activity of Methanolic Extract and Isolated Compound of Pavonia odorata Roots in Mice. Research Journal of Pharmacy and Technology. 2021; 14(7): 3489-4. doi: 10.52711/0974-360X.2021.00605.

7.      Suhailah Hayaza, Win Darmanto, Sri Puji Astuti Wahyuningsih, Raden Joko Kuncoroningrat Susilo, Saikhu Akhmad Husen, Dwi Winarni, Ruey-an Doong. Immunomodulatory Activity of Okra Raw Polysaccharide Extract by Regulating TNF-A, IFN-G Levels, and Cell Apoptosis in DEN-induced mice. Research Journal of Pharmacy and Technology. 2022; 15(2): 546-0. doi: 10.52711/0974-360X.2022.00088

8.      Bhokare P, Khadke A, Kuchekar G and Kulkarni S. Comparative study of different extraction technique and phytochemical screening of Delonixregia. J Pharmacogn Phytochem 2018;7(4):133-8.

9.      Kumar AR, Shaik R, Yeshwanth D. Phytochemical evaluation of Delonixregia, Samaneasaman, Bauhinia variegatga. International Journal of research in Pharmacy and Chemistry. 2013; 3(4): 768-72.

10.   Dr. Khandelwal K R . Practical Pharmacognosy, 22nd edition, Nirali Prakashan Pune 2012.

11.   Shewale VD, Deshmukh TA, Patil LS, Patil VR. Anti-Inflammatory Activity of Delonixregia (Boj. Ex. Hook). Adv Pharmacol Sci. 2012; 174-85.

12.   Ahmed J, Nirmal S, Dhasade V, Patil A, Kadam S, Pal S, Mandal S, Pattan S. Hepatoprotective activity of methanol extract of aerial parts of Delonixregia. Phytopharmacology. 2011; 1(5): 118-22.

13.   Dashputre N.L and Naikwade N.S. Immunomodulatory activity of Abutilon indicum Linn. On Albino Mice. IJPSR. 2010; 1(3): 178-84.

14.   Tripathi S, Maurya A.K, Kahrana M, Kaul A and Sahu R.K. Immunomodulatory property of ethanolic extract of Trigonellafoenum-graeceum leaves on mice. Scholar Research Library. 2012; 4(2): 708-13.

15.   Sharma A, Rangari V. Immunomodulatory activity of methanol extract of Adansonia digitata L. Trop J Pharm Res. 2016; 15(9): 1923-27

16.   Savant C, Joshi N, Reddy S, Mannasaheb BA, Joshi H. Immunomodulatory medicinal plants of India: A review. Int. J. Pharma. Toxicol. 2014; 1(4): 109-15.

17.   Ganeshpurkar A, Saluja AK. Experimental animal models used for evaluation of potential immunomodulators: A mini review. Bulletin of Faculty of Pharmacy, Cairo University. 2017; 55(2): 211-16.

18.   Shukla S and Mehta A. Comparative phytochemical analysis and in vivo immunomodulatory activity of various extracts of Stevia rebaudiana leaves in experimental animal model. Frontiers in Life Science. 2015; 8(1): 55-63.

19.   Sneha J Anarthe, Sunitha D. Sandhyarani. Immunomodulatory activity of methanolic extract of Trigonellafoenum-graecum whole plant in Wistar albino rat. American Journal of Phytomedicine and Clinical Therapeutics. 2014; 2 (9): 1081-92.

20.   Ghase M, Devkate R, Fargade D. The flamboyant delonixregia (Gulmohar). International Journal of Creative Research Thoughts. 2022; 10(3): 586-93

21.   Ajith Y, Dimri U, Dixit SK, Singh SK, Gopalakrishnan A, Madhesh E, Rajesh JB, Sangeetha SG. Immunomodulatory basis of antioxidant therapy and its future prospects: An appraisal. Inflammopharmacology. 2017; 25(5): 487-98.

22.   Patel P, Asdaq SM. Immunomodulatory activity of methanolic fruit extract of Aeglemarmelos in experimental animals. Saudi Pharm J. 2010; 18(3):161-5.

23.   Nyi Mekar Saptarini, Driyanti Rahayu, Endah Kartikawati. Immunomodulatory Activity of Crude Bromelain of Pineapple (Ananas comosus (L.) Merr.) Crown from Subang District, Indonesia. Research J. Pharm. and Tech. 2020;  13(11): 5177-5182. doi: 10.5958/0974-360X.2020.00905.1

24.   Shantilal Singune, Jai S Vaghela. Investigation of Immunomodulatory Activity of Methanolic Extract and Isolated Compound of Pavonia odorata Roots in Mice. Research Journal of Pharmacy and Technology. 2021; 14(7): 3489-4. doi: 10.52711/0974-360X.2021.00605

25.   Sudha P, Asdaq SM, Dhamingi SS, Chandrakala GK. Immunomodulatory activity of methanolic leaf extract of Moringaoleifera in animals. Indian J Physiol Pharmacol. 2010; 54(2): 133-40.

26.   Janeway CA, Travers P, Walport JR, Shlomchik, MJ. The immune system in health and disease: immunobiology. 5. New York: Garland Publishing. 2001: 1-312.

 

 

 

 

 

 

Received on 22.11.2022            Modified on 07.03.2023

Accepted on 21.06.2023           © RJPT All right reserved

Research J. Pharm. and Tech 2024; 17(3):1296-1302.

DOI: 10.52711/0974-360X.2024.00203