Adryan Fristiohady1, Taifo Mahmud2, Nuralifah1, Loly Subhiaty Idrus1,
La Ode M. J. Purnama3, Lidya A Haruna3, Tyaz Andarzia1, Fery I. Armadany1,
Fadhliyah Malik1, Rathapon Asasutjarit3, Monsicha Khuanekkaphan3, Idin Sahidin1,
Wahyuni Wahyuni1
1Faculty of Pharmacy, Universitas Halu Oleo, Kendari 93232, Indonesia.
2Department of Pharmaceutical Sciences, Oregon State University,
Corvallis, Oregon 97331-3507, United States of America.
3Thammasat University Research Unit in Drug, Health Product Development, and Application (DHP-DA), Department of Pharmaceutical Sciences, Faculty of Pharmacy.
*Corresponding Author E-mail: adryanfristiohady@uho.ac.id
ABSTRACT:
This study aims to evaluate the anti-inflammatory activity of Lycopene in mice with ear oedema and in vitro activity. In this study, mice were used and performed in vitro anti-inflammatory tests of Lycopene with the HRBC method and protein denaturation. Moreover, the anti-inflammatory assay of Lycopene was also evaluated in xylene-induced right-ear mice. Diclofenac sodium solution was used as a positive control. The results showed that Lycopene provides anti-inflammatory activity in vitro by stabilizing the human red blood cell (HRBC), preventing lysis, and inhibiting protein denaturation with IC50 32.47 μg/ml. In addition, Lycopene also reduced the thickness of xylene-induced oedema in the right ear of mice. Lycopene provides anti-inflammatory properties by inhibiting HRBC lysis and protein denaturation and stabilizing HRBC, decreased the thickness of Oedema in the right ear of mice and the TNF-⍺ levels.
KEYWORDS: Lycopene, Anti-Inflammatory, HRBC, Protein denaturation, Oedema.
INTRODUCTION:
Inflammation is a local reaction of an area of the body to tissue damage characterized by symptoms such as swelling, fever, and pain. Inflammation can be classified as either acute or chronic. During acute inflammation, immune cells eliminate viruses or other pathogens through various mechanisms. In the context of a sustained or repeated inflammatory response, various cytokines are secreted to impair macrophage function.
Several inflammatory cytokines and enzymes play roles in inflammation responses, such as tumour necrosis factor α (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS)1,2,3,4. The cytokines interleukin-6 (IL-6) and TNFα are increased in most inflammatory settings and have been recognized as targets for therapeutic interventions5. IL-6 is a soluble mediator with pleiotropic effects on inflammation, immune response, and haematopoiesis6. Whereas Tumour necrosis factor-⍺ (TNF-⍺, also known as cachectin) is a potent proinflammatory cytokine that plays a vital role in the immune system during inflammation, cell proliferation, differentiation, and apoptosis7. Inflammatory effects can be a severe problem if not handled properly. Chronic inflammation, including metabolic syndrome, type 2 diabetes, non-alcoholic fatty liver disease (NAFLD), cardiovascular disease, cancer, depression, autoimmune diseases, neurodegenerative diseases, sarcopenia, osteoporosis, and immune sensitization could cause complicated problems8.
Currently, non-steroidal anti-inflammatory drugs (NSAIDs) are used for inflammatory diseases. These drugs temporarily suppress inflammation, inhibition of cyclooxygenase enzymes which are responsible for the conversion of Arachidonic acid to prostaglandins, but their long-term use causes side effects such as gastrointestinal tract ulceration and renal morbidity. So, research is focused on finding new drugs with pharmacological actions without side effects9,2,10.
Lycopene, a carotenoid compound that is the main component of red fruits and vegetables such as tomatoes and other fruits, is composed of 40 carbon atoms with an acyclic open chain structure connected via unsaturated and conjugated double bonds, 13 double bonds with 11 conjugated double bonds and two unconjugated double bonds. This distinctive conjugated polyene structure gives Lycopene its bright red colour and antioxidant properties. Lycopene is a potent protector against oxidative damage to DNA, proteins, and lipids, reduces cardiovascular disease risk factors, and has anticancer, antioxidant, antibacterial, immunomodulatory, wounds healing, and anti-inflammatory effects1,11,12,13,14,15. Most studies point to the anti-inflammatory effect of Lycopene due to its ability to modulate the pathways responsible for the induction of inflammatory mediators16.
Lycopene from Chlorella marina showed 83% inhibition in carrageenan-induced male Sprague Dawley rat leg oedema. Lycopene showed decreased activity of cyclooxygenase (COX) and lipoxygenase (LOX) in monocytes, decreased Myeloperoxidase (MPO) in serum, C-reactive protein (CRP), and ceruloplasmin activity in plasma9. Another study demonstrated that Lycopene significantly inhibits the formation of leg oedema at two doses (25 and 50mg/kg) in acute and repeated administration. In addition, Lycopene exhibits local anti-inflammatory activity and attenuates liver damage caused by I/R. Mice underwent ischemia for 45 minutes in three-quarters of the liver, followed by reperfusion for 2 hours. In this model, Lycopene was administered daily in two doses (25 and 50mg/kg) for a 14-day trial17. Lycopene in watermelon exhibits very high antioxidant activity and superoxide anion scavenging activity comparable to Trolox. A lycopene from watermelon at dose (100µg/mL) inhibited the expression of iNOS and COX-2 mRNA levels and their proteins, indicating solid anti-inflammatory activity. Thus, lycopene is a good source of antioxidants and anti-inflammatory agents18, protects against oxidative damage to lipids, proteins and DNA13. Lycopene also restrained the activation of NF-κB and JNK, which cause inflammation, and suppressed the expression of TNF-α, IL-1β, IL-6, COX-2, and iNOS in SW480 human colorectal cancer cells1. However, there is no data reported the anti-inflammatory data from Lycopene to inhibits the ear oedema and compare with the in-vitro analysis. According to the descriptive, this study aims to test the anti-inflammatory activity of Lycopene in mice with ear oedema and in vitro evaluation.
MATERIALS AND METHODS:
Materials:
Lycopene was obtained from Shaanxi Jintai Biological Engineering Co,. Ltd. Buffer phosphate pH 7.4, diclofenac sodium, topical xylene, bovine serum albumin (BSA), tris base, sodium chloride (NaCl), and sodium carboxymethylcellulose (Na-CMC) were used in this study. All chemicals used were analytical grade.
Animals:
In this study, mice (20g) were used. The mice were acclimatized for 7 d prior experiment and maintained in the cage at 25°C with a 12 h light/dark cycle. All experiment involving animals was conducted according to ethical clearance no. 146b/UN29.20.1.2/PG/2023 declared by Research Ethical Committee, Halu Oleo University, Indonesia.
In vitro Anti-inflammatory of Lycopene:
Anti-inflammatory of Lycopene in HRBC:
Blood used for the HRBC method was collected from healthy volunteers. Then, the blood was centrifuged for 10 min at 3,000 rpm at room temperature. The supernatant was collected and washed with isosaline solution. Later, the 10% v/v isosaline red blood cell suspension was obtained for an anti-inflammatory test with HRBC method. The 10% v/v isosaline red blood cell suspension was mixed with phosphate buffer pH 7.4 (1 ml), hyposaline solution (2 ml), 10% v/v isosaline red blood cell suspension (0.5 ml), and 1 ml either of lycopene (Lyc) at different concentration and diclofenac sodium solution for sample test and positive control, respectively. The negative control was also prepared without adding the sample test or diclofenac sodium solution. All mixtures were incubated at 56°C for 30 min and continued by centrifugation at 5,000 rpm for 10 min. The supernatant was collected, and the haemoglobin content was measured with a UV-vis spectrophotometer at λ560 nm. The haemolysis and stability percentage were calculated as follows:19
% Hemolysis = (Sample Absorbance/Negative Control Absorbance) x 100 (1)
% Stability = (Sample Absorbance/Negative Control Absorbance) x 100 (2)
Anti-inflammatory of Lycopene on Protein Denaturation:
Protein denaturation is one of the in vitro analyses for determining anti-inflammatory. Briefly, 50μl of diclofenac sodium solution and sample solution were added into 5ml of 0.2% v/v BSA (bovine serum albumin) in TBS (tris buffer saline). The mixtures were then incubated at 25°C for 30 minutes, continued at 72°C for a minute, and cooled at 25°C for 25 minutes. Then, the absorbance was measured with a UV-vis spectrophotometer at λ 660nm. The inflammatory activity was calculated by estimating the protein denaturation inhibition, calculated as follows:
% Inhibition = [(Negative Control Absorbance - Sample Absorbance)/Negative Control Absorbance] x 100
Anti-Inflammatory Effects of Lycopene on Xylene-Induced Ear Oedema:
Thirty mice were randomly divided into six groups, which were the sham group (normal control), naďve group (administered 0.5% of Na-CMC), positive group (administered 0.2mg of diclofenac sodium), 0.025, 0.05, and 0.1μg/ml of lycopene, respectively. On day 8, animals were induced with topical xylene for inflammatory response. Later, the right ear thickness of mice was measured with an ultrasonic thickness gauge meter at min-15, 30, and 45, respectively. After 45 minutes, the mice were sacrificed, and their blood was collected for further analysis20.
The inflammatory percentage was calculated as follows:
% Inflammatory = [(Dt – D0)/D0] x 100 (1)
Dt is the ear thickness at the indicated time (t), and D0 is the ear thickness at min-0. Then, the inflammatory inhibition was calculated by:
% Inhibition = [(a – b) / a] x 100 (2)
Whereas a is % inflammatory in the sham group, and b is %inflammatory in treatment group.
Statistical Analysis:
Data collected was analysed using GraphPad Prism 9. The descriptive data were expressed as mean±standard error of the mean. The anti-inflammatory activity was analysed by ANOVA test with a CI of 95%. The difference was considered significant if the p-value was less than 0.05.
RESULT:
Lycopene (Lyc) is one of the natural products that is mainly isolated from tomatoes. Lycopene has broad pharmacological activities, including antioxidant, anti-mutagenic, anticancer, anti-inflammatory, and anti-atherogenesis21.
The human red blood cells (HRBC) method is an in vitro assay for estimating the anti-inflammatory properties of samples22,23. This assay allows the characterization of the capacity of Lyc to protect erythrocytes from haemolysis when heat is supplied24. The erythrocyte membrane mimics the lysosomal membrane, thereby the ability of samples to stabilize the erythrocyte membrane might mimic the membrane to release activated neutrophils from lysosomal components, which leads further tissue inflammation and damage22. The results found that Lyc could stabilize human red blood cells (HRBC) and prevent HRBC lysis. Most anti-inflammatory drugs exert beneficial effects by inhibiting either the release of lysosome enzymes or by stabilizing the lysosome membrane, which is one of the principals responsible for the inflammatory process25,26,10, so Lyc has the potential to be used as an anti-inflammatory drug that inhibits lysosomal enzymes or stabilizes their membrane24. The result is shown in Figure 1 and Table 1.
Table 1. The % haemolysis and % stability of Lyc and Na-Dic.
|
Concentration (µg/mL) |
Lyc |
Na-Dic |
|||||||||||
|
Hemolysis (%) |
Stability (%) |
Hemolysis (%) |
Stability (%) |
||||||||||
|
Mean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
|
|||||
|
Lower Bound |
Upper Bound |
Lower Bound |
Upper Bound |
Lower Bound |
Upper Bound |
Lower Bound |
Upper Bound |
|
|||||
|
10 |
41.00 |
39.89 |
42.12 |
59.00 |
57.88 |
60.11 |
46.17 |
45.31 |
47.03 |
53.83 |
52.97 |
54.69 |
|
|
20 |
35.99 |
34.30 |
37.68 |
64.01 |
62.32 |
65.70 |
41.16 |
39.84 |
42.47 |
58.84 |
57.53 |
60.16 |
|
|
30 |
31.61 |
30.85 |
32.38 |
68.39 |
67.62 |
69.15 |
36.62 |
35.20 |
38.05 |
63.38 |
61.95 |
64.80 |
|
|
40 |
24.73 |
24.28 |
25.18 |
75.27 |
74.82 |
75.72 |
27.70 |
26.86 |
28.54 |
72.30 |
71.46 |
7315 |
|
|
50 |
17.06 |
15.14 |
18.98 |
82.94 |
81.02 |
84.86 |
18.31 |
15.18 |
21.44 |
81.69 |
78.56 |
84.82 |
|
|
60 |
9.39 |
8.17 |
10.61 |
90.61 |
89.39 |
91.83 |
10.95 |
9.30 |
12.61 |
89.05 |
87.39 |
90.70 |
|
Data is presented as mean with CI 95%
Figure 1. The haemolysis and stability activity of Lycopene and Diclofenac sodium in human red blood cells (HRBC). Data is presented as mean with CI 95%
Lyc also has an anti-inflammatory by inhibiting the denaturation protein. It correlates with the inflammatory response to stimuli. The protein denaturation might lead to tissue injury27 and the production of autoantigens in inflammation conditions. Inhibition of protein denaturation can be inhibited inflammatory activity20,28. Thereby, the anti-inflammatory activity in vitro with protein denaturation inhibition might be the potential for evaluating the anti-inflammatory activities of the sample27. According to the results (Table 2 and Figure 2), IC50 of Lyc and Na-Dic was found to be 32.47 and 11.66μg/ml. Lycopene (Lyc) could inhibit protein denaturation, although Na-Dic significantly inhibited the protein denaturation.
The results showed that the administration of Lyc 25, 50, and 100 reduced the thickness of mice induced with xylene (Table 3 and Figure 3). The higher concentration of Lycopene showed better efficacy in decreasing the oedema, and they are comparable with the positive control used. Lyc might work as anti-inflammation in acute inflammation due to its ability to reduce oedema less than 30 min induced by inhibiting the release of substance P (SP). Xylene releases SP, secreted by nerve and inflammatory cells, including macrophages, eosinophils, lymphocytes, and dendritic cells, by binding to NK-1R (neurokinin-1 receptor). It increases the capillary permeability and leukocyte infiltration, leading to Oedema29,30.
Table 2. The protein denaturation inhibitory activity of Lyc
|
Concentration (µg/mL) |
Inhibition (%) |
IC50 (µg/ mL) |
||
|
Na-Dic |
Lyc |
Na-Dic |
Lyc |
|
|
10 |
50.281±0.117 |
35.288±0.205 |
11.66 |
32.47 |
|
20 |
52.983±0.204 |
36.664±2.261 |
||
|
30 |
54.564±0.189 |
50.433±0.044 |
||
|
40 |
56.706±0.158 |
53.646±0.190 |
||
|
50 |
59.051±0.262 |
56.859±0.187 |
||
|
60 |
60.938±0.267 |
59.918±0.035 |
||
|
70 |
62.978±0.182 |
61.397±0.077 |
||
Figure 2. The inhibitory concentration of Lyc (A) and Na-Dic (B) in inhibiting the protein denaturation
Table 3. %oedema and %inhibition of Lyc in xylene-induced right ear mice. Data is presented as mean with CI 95%
|
Group |
Oedema (%) |
||||||||
|
15 min |
30 min |
45 min |
|||||||
|
ean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
||||
|
Lower Bound |
Upper Bound |
Lower Bound |
Upper Bound |
Lower Bound |
Upper Bound |
||||
|
Control (-) |
321.78 |
297.27 |
356.28 |
470.95 |
454.06 |
487.83 |
633.52 |
582.46 |
684.57 |
|
Control (+) |
280.51 |
252.50 |
308.51 |
143.54 |
131.17 |
155.91 |
5.67 |
5.08 |
6.26 |
|
Lyc 25 |
296.49 |
220.45 |
372.53 |
235.30 |
184.31 |
286.28 |
136.41 |
114.55 |
158.28 |
|
Lyc 50 |
288.63 |
202.13 |
375.14 |
195.64 |
103.41 |
287.86 |
73.32 |
59.59 |
87.05 |
|
Lyc 100 |
272.18 |
257.19 |
287.17 |
151.38 |
147.92 |
154.83 |
14.40 |
11.36 |
17.43 |
Continue Table 3
|
Group |
Inhibition (%) |
||||||||
|
15 min |
30 min |
45 min |
|||||||
|
Mean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
Mean |
95% Confidence Interval for Mean |
||||
|
Lower Bound |
Upper Bound |
Lower Bound |
Upper Bound |
|
Lower Bound |
Upper Bound |
|||
|
Control (-) |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
Control (+) |
12.71 |
-1.90 |
34.04 |
69.51 |
65.81 |
73.20 |
99.10 |
98.96 |
99.25 |
|
Lyc 25 |
7.74 |
-18.55 |
42.36 |
50.01 |
38.01 |
62.00 |
78.44 |
73.90 |
82.97 |
|
Lyc 50 |
10.10 |
-22.16 |
17.49 |
58.44 |
38.55 |
78.34 |
88.41 |
85.47 |
91.34 |
|
Lyc 100 |
15.40 |
13.31 |
13.98 |
67.85 |
67.06 |
68.65 |
97.72 |
97.07 |
98.37 |
Figure 3. The thickness of ear right of xylene-induced oedema in mice. Data is measured at min-15, 30, and 45. Data is presented as mean with CI 95%
CONCLUSION:
Lycopene provides anti-inflammatory properties in vitro by inhibiting human red blood cell (HRBC) lysis and protein denaturation and stabilizing HRBC. In addition, it also decreased the thickness of xylene-induced Oedema in the right ear of mice and decreased the TNF-⍺ levels.
CONFLICT OF INTEREST:
Authors declared there is no conflict of interest.
FUNDING:
This research has been funded by The internal research grant of Faculty of Pharmacy, Halu Oleo University 2023.
ACKNOWLEDGMENTS:
Thank you to the World Class Professor 2023 Program by Directorate of Resources Affairs, Directorate General of Higher Education, Research, and Technology, Ministry of Education, Culture, Research and Technology, Republic of Indonesia.
REFERENCES:
1. Cha JH, Kim WK, Ha AW, Kim MH, Chang MJ. Anti-inflammatory effect of Lycopene in SW480 human colorectal cancer cells. Nutrition Research and Practice, 2017; 11(2): 90-96.
2. Dhalendra G, Satapathy T, Roy A. Animal models for inflammation: A review. Asian Journal of Pharmaceutical Research. 2013; 3(4): 207-212.
3. Mamillapalli V, Chapala RH, Sareddu TKS, Kondaveeti LS, Pattipati S, Khantamneni P. Evaluation of phytochemical and in vitro anti-inflammatory activity of leaf and fruit extracts of Casuarina equisetifolia. Asian Journal of Pharmacy and Technology. 2020; 10(3): 143-148.
4. Ganer R, Kamble MA, Dhabarde DM, Ingole AR, Baheti JR. Evaluation of Analgesic and In-vitro Anti-Inflammatory potential of Fruit Flesh Extract of Terminalia catappa Linn. Research Journal of Pharmacognosy and Phytochemistry. 2017; 9(4): 228-230.
5. Scheller J, Chalaris A, Schmidt-Arras D, Rose-John S. The pro-and anti-inflammatory properties of the cytokine interleukin-6. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research. 2011; 1813 (5): 878-888.
6. Tanaka T, Narazaki M, Kishimoto T. IL-6 in inflammation, immunity, and disease. Cold Spring Harbor perspectives in biology.2014; 6(10): a016295
7. Zelová H, Hošek J. TNF-α signalling and inflammation: interactions between old acquaintances. Inflammation Research. 2013; 62(7): 641-651
8. Furman D, Judith C, Eric V, Pedro CB, Sasha T, Claudio F, Luigi F, Derek WG, Alessio F, Gary WM, Adrew HM, Alberto M, Cornelia MW, Nir B, Jorg JG, Thimas AR, Rita BE, Alejandro L, Nicole K, George MS. Chronic Inflamation in the Etiology of Deases Across the Life Span. Natural Medicines. 2020; 25(12): 1822–1832.
9. Renju GL, Kurup GM, Saritha KCH. Effect of lycopene from Chlorella marina on high cholesterol-induced oxidative damage and inflammation in rats. Inflammopharmacology. 2013; 22: 45-54.
10. Siju P, Ghetia R, Vadher B, Manvar MN. In-Vitro Anti-inflammatory Activity of Fractions of Ailanthus excelsa Roxb. by HRBC Membrane Stabilization. Asian Journal of Pharmacy and Technology. 2015; 5(1): 29-31.
11. Kim SO, Ha TVA, Choi YJ, Ko S. Optimization of homogenization–evaporation process for lycopene nanoemulsion production and its beverage applications. Journal of Food Science. 2014; 79(8): N1604-N1610.
12. Yaping Z, Wenli YU, Weile HU, Ying Y. Anti-inflammatory and anticoagulant activities of Lycopene in mice. Nutrition Research. 2003; 23 (11): 1591-1595.
13. Sheriff SA, Devaki, T. Effect of Lycopene on general clinical parameters during D-galactosamine/Lipopolysaccharide (D-GalN/LPS) induced hepatitis in Rats. Research Journal of Pharmacy and Technology. 2012; 5(3): 398-403.
14. Sahana K. Lycopene as an antioxidant and its medicinal uses. Research Journal of Pharmacy and Technology. 2015; 8(8): 1043-1047.
15. Chopra M, Bhaumik A, Reddy AG, Kumar PS, Srikant B. Extraction and Isolation of Bioactive Molecule Lycopene from Water Melon and Evaluation of Anti Diabetic Activity against STZ Induced Rats. Research Journal of Pharmacy and Technology. 2018; 11(1): 101-106.
16. Moia VM, Leal PF, Almeida PT, Barbosa CL, Ricci-Junior E, Cruz RE, Magalhaes RAL, Savio MSF, Sampson A, Hussain IS, Alexis F, de OliveiraHenriques MD, Santos-Oliveira R. Lycopene used as Anti-inflammatory Nanodrug for the Treatment of Rheumathoid Arthritis: Animal assay, Pharmacokinetics, ABC Transporter and Tissue Deposition. Colloids and surfaces. B, Biointerfaces. 2020; 188: 110814.
17. Bignotto L, Rocha J, Sepodes B, Eduardo-Figueira M, Pinto R, Chaud M, Mota-Filipe H. Anti-inflammatory effect of Lycopene on carrageenan-induced paw oedema and hepatic ischaemia–reperfusion in the rat. British Journal of Nutrition. 2009; 102(1): 126-133.
18. Kim CH, Park MK, Kim SK, Cho YH. Antioxidant capacity and anti‐inflammatory activity of Lycopene in watermelon. International Journal of Food Science and Technology. 2014; 49 (9): 2083-2091.
19. Yesmin S, Paul A, Naz T, Rahman ABM, Akhter SF, Wahed MII, Emran TB, Siddiqui SA. Membrane stabilization as a mechanism of the anti-inflammatory activity of ethanolic root extract of Choi (Piper chaba). Clin Phytosci 2020; 6(59).
20. Dharmadeva S, Galgamuwa LS, Prasadinie C, Kumarasinghe N. In vitro anti-inflammatory activity of Ficus racemosa L. bark using albumin denaturation method. Ayu. 2018; 39(4): 239–242.
21. Chaudhary P, Sharma A, Singh B, Nagpal AK. Bioactivities of phytochemicals present in tomato. Journal of Food Science and Technolog. 2018; 55(8): 2833–2849
22. Saleem TK, Azeem AK, Dilip C, Sankar C, Prasanth NV, Duraisami R. Anti-inflammatory activity of the leaf extacts of Gendarussa vulgaris Nees. Asian Pacific Journal of Tropical biomedicine. 2011; 1(2): 147-149
23. Jaiganesh KP, Jasna TJ, Tangavelou AC. Phytochemical, In vitro anti-inflammatory and antimicrobial potential of Hugonia mystax L. Research Journal of Pharmacognosy and Phytochemistry. 2021; 13(4): 169-173.
24. Amaro HM, Barros R, Tavares T, Almeida R, Pinto IS, Malcata FX, Guedes AC. Gloeothece sp.—Exploiting a New Source of Antioxidant, Anti-Inflammatory, and Antitumor Agents. Marine Drugs. 2021; 19(11): 623
25. Amabye TG. Antioxidant and anti-inflammatory properties of cultivated mushrooms grown in mekelle city Tigray Ethiopia. International Journal Nutrition Food Sciences. 2015; 4(5): 578-583.
26. Gorla US, Savithri M, Rao GSN, Niharika Y, Sathya P, Harika V. Evaluation of anti-inflammatory activity of Hydroalcoholic extract of Ananas cosmosus fruit peel by HRBC membrane stabilisation. Asian Journal of Pharmaceutical Research. 2018; 8(1): 33-35.
27. Osman NI, Sidik NJ, Awal A, Adam NA, Rezali NI. In vitro xanthine oxidase and albumin denaturation inhibition assay of Barringtonia racemosa L. and total phenolic content analysis for potential anti-inflammatory use in gouty arthriti. Journal of Intercultural Ethnopharmacology. 2016; 5(4): 343–349.
28. Thawkar B, Kale M, Oswal M, Maniyar K, Kadam K, Kamat S. To study anti-inflammatory activity of 70% methanolic Extract of Triumfetta rhomboidea: In vitro Study. Research Journal of Pharmacy and Technology. 2016; 9(3): 241-244.
29. Singsai K, Charoongchit P, Chaikaew W, Boonma N, Fhanjaksai P, Chaisatan K. Antilipoxygenase and Anti-Inflammatory Activities of Streblus asper Leaf Extract on Xylene-Induced Ear Edema in Mice. Advances in Pharmacological and Pharmaceutical Sciences. 2020
30. Sun K, Song X, Jia R, Yin Z, Zou Y, Li L, Yin L, He C, Liang X, Yue G, Cui Q,Yang Y. Evaluation of Analgesic and Anti-Inflammatory Activities of Water Extract of Galla Chinensis In Vivo Models. Evidence-based complementary and alternative medicine:eCAM.2018.
|
Received on 06.03.2024 Revised on 25.07.2024 Accepted on 31.10.2024 Published on 27.03.2025 Available online from March 27, 2025 Research J. Pharmacy and Technology. 2025;18(3):1052-1057. DOI: 10.52711/0974-360X.2025.00151 © RJPT All right reserved
|
|
|
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License. |
|