GANODERMA LUCIDUM TOTAL TRITERPENES ALLEVIATE INFLAMMATION AND MODULATE ANTIOXIDANT STATUS IN FREUND’S COMPLETE ADJUVANT-INDUCED ARTHRITIC RATS

GANODERMA LUCIDUM TOTAL TRITERPENES ALLEVIATE INFLAMMATION AND MODULATE ANTIOXIDANT STATUS IN FREUND’S COMPLETE ADJUVANT-INDUCED ARTHRITIC RATS

T. P. Smina, J. Mathew, K. A. Rony and K. K. Janardhanan ^*

Amala Cancer Research Centre, Thrissur - 680555, Kerala, India.

ABSTRACT: Ganoderma lucidum total triterpenes were evaluated for anti-inflammatory activity using carrageenan-induced acute and formalin-induced chronic paw oedema models in Swiss albino mice. The total triterpenes, at concentrations 10, 50, and 100 mg/kg b. wt. administered orally, showed 36.96%, 54.35%, 76.09% inhibition of acute and 41.86%, 62.79% and 79.07% inhibition of chronic inflammation. Total triterpenes were also assessed for its effect in attenuating the inflammation and modulating the antioxidant status of Freund’s complete adjuvant (FCA) induced arthritis rats. Total triterpenes when administrated orally, at concentrations 10, 50, and 100mg/kg b. wt., showed 58.73%, 73.02%, and 79.37% inhibition of paw oedema in the arthritis rats. Treatment with total triterpenes enhanced the activities of the antioxidant enzymes (SOD, GPx, and catalase) and restored the level of GSH in arthritic animals. Total triterpene treatment also attenuated the enhanced lipid peroxide levels in FCA-induced rats. All these findings support the strong therapeutic potential of Ganoderma total triterpenes against inflammation associated-diseases.

Anti-inflammatory, Anti-arthritic, Oedema, Carrageenan, formalin

INTRODUCTION: Inflammation is a defensive response for eliminating the harmful agent and injured tissues, thereby stimulating tissue repair. When this critical and usually beneficial response occurs in an uncontrolled manner, the result is excessive cellular damage that results in chronic inflammation and the destruction of healthy tissue. Inflammation has been associated with the patho physiology of numerous clinical conditions, including vascular diseases and cancer ^{1, 2}.

The term arthritis is used to define several forms of painful degenerative and inflammatory joint diseases ³. Rheumatoid arthritis is a poly-articular joint disease characterized by massive synovial proliferation, sub-intimal infiltration of inflammatory cells, and subsequent destruction of cartilage and bone ⁴. It can affect multiple organs, including muscle, bone, and other soft tissues, thus causing varying degrees of joint deformity and instability, leading to painful disabling conditions ³.

Anti-inflammatory and anti-arthritic agents exert various effects that result in the regular activity of the immune system, including the reduction of the number of immune cells. Several natural products are being used as excellent anti-inflammatory agents without worrying about potential side effects. Extracts of G. lucidum were found to possess a significant anti-inflammatory effect against different inflammatory models in animals ^{5, 6, 7}. Earlier studies conducted in our laboratory also revealed the anti-peroxidative, anti-inflammatory, and anti-mutagenic activities of ethanol extract of the mycelium of Ganoderma lucidum ⁸. Ganoderma lucidum polysaccharide was found to show significant activity in carrageenan-induced acute and formalin-induced chronic inflammation ⁹. Ganoderma lucidum polysaccharide was also found to influence the production of pro-inflammatory cytokines in activated rheumatoid synovial fibroblasts ¹⁰. The present work evaluated the anti-inflammatory activity of total triterpenes isolated from Ganoderma lucidum using carrageenan-induced acute and formalin-induced chronic paw oedema models. An attempt is also made to assess the effect of total triterpenes in attenuating the inflammation and modulating the antioxidant status of Freund’s complete adjuvant (FCA) induced arthritis rats.

MATERIALS AND METHODS:

Isolation of Total Triterpenes: Isolation of the total triterpenes from the fruiting bodies of G. lucidum was performed as described earlier ¹¹. Briefly, a chloroform soluble fraction from the 100% ethanolic extract of G. lucidum fruiting bodies was separated and concentrated. The concentrate was then loaded onto a silica gel column and eluted with petroleum ether, chloroform, methanol, and various combinations of these solvents. The fractions that answered the tests for triterpenes 12 were combined and concentrated to give the total triterpenes.

Animals: Male Swiss albino mice used for anti-inflammatory studies and female Wistar rats used for anti-arthritic studies were purchased from Small Animal Breeding Station, Mannuthy, Kerala, India, and were housed in well-ventilated cages under controlled conditions of light and humidity. Animals were provided with standard mouse chow (Sai Durga Food and Feeds, Bangalore, India) and water ad libitum. All the animal experiments were carried out as per the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Environment and Forest, Government of India, and by the approval of the Institutional Animal Ethical Committee (149/99/CPCSEA dated 23-10-2009).

Anti-Inflammatory Activity of Total Triterpenes:

Carrageenan-Induced Acute Paw Oedema: Male Swiss albino mice weighing 25 ± 2 g (6 weeks old) were divided into five groups of 6 animals. Acute inflammation was produced in all animals by the sub-plantar injection of 20 µl of freshly prepared 1% suspension of carrageenan in normal saline on the right hind paw ^{13, 14}. Group 1 comprises control animals that received only carrageenan treatment. Animals in Groups 2, 3, and 4 were pre-medicated with total triterpenes (10, 50, and 100 mg/kg body weight) and group 5 with the reference drug diclofenac (10 mg/kg body wt.), orally one hour before carrageenan injection. Group 6 animals were treated with 250μl sunflower oil which was used as the vehicle to administer total triterpenes. The paw thickness was measured using Vernier calipers before and three hours after the carrageenan challenge, from which the degree of oedema formation was calculated.

Formalin-Induced Chronic Paw Oedema: Male Swiss albino mice weighing 25 ± 2 g (6 weeks old) were treated in the same way as in the case of carrageenan-induced paw oedema model ¹⁵. Instead of carrageen, 20 µl of freshly prepared 2% formalin was used as the oedema to the genic agent. Diclofenac (10 mg/kg body weight) was used as the reference drug. The drug treatment was continued for six consecutive days. The paw thickness and the degree of oedema formation were determined regularly for six days after the formalin injection.

Determination of Anti - Arthritic Activity using FCA Induced Arthritis Model: Female Wistar rats weighing 180 ± 20g (15 weeks old) were divided into six groups, consisting of six animals. Arthritis was induced in all groups, except group 1, by the intradermal injection of 0.1ml of Freund’s Complete Adjuvant (Genei, Bangalore) into the sub-planar region of the right hind paw ^{16, 17}.

Group 1 include healthy animals that received neither the adjuvant injection nor the drug treatment. Group 2 include control animals that were inoculated with Freund’s complete adjuvant but received no drug treatment. Group 3 animals were administered with standard reference drug diclofenac orally at a dosage of 10mg/kg body weight in 0.5 ml distilled water.

Group 4, 5, and 6 animals received 10, 50, and 100 mg/kg body weight total triterpenes orally in 250 μl sunflower oil. Group 7 animals received 250 μl sunflower oil that was used as the vehicle to administer total triterpenes. Diclofenac, total triterpenes, and the vehicle were administered orally one hour before Freund’s complete adjuvant (FCA) injection. The oral administration of the total triterpenes and the vehicle were continued, once daily, for 12 days.

The change in paw thickness was assessed by measuring the right hind paw volume using A Vernier calipers just before drug administration and after adjuvant injection. The paw thickness measurement was repeated every three days after adjuvant inoculation until 22^nd day. On the 22^nd day, i.e., at the end of the experimental period, the severity of the secondary lesions was evaluated visually and scored.

The animals were killed under anesthesia and blood samples were collected directly from the heart, and the non-coagulated (heparinised) blood was used for the determination of antioxidant enzyme activities and reduced glutathione level. The serum samples, after precipitating the protein ¹⁸, were used for determining the lipid peroxidation levels ¹⁹. The whole blood was used for analyzing Hb content 20, and the erythrocyte lysate was used to estimate the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH). The activity of SOD in blood was assessed, after removing the haemoglobin, by the method of Mc Cord and Fridovich ²¹. Catalase activity in the blood was measured by the method of Aebi 22, and the glutathione peroxidase activity was determined according to the method of Hafemann et al. ²³. Reduced glutathione in blood was determined, after preparing a hemolysate in water, according to the method of Moron et al. ²⁴.

Statistical Analysis: All values are expressed as mean ± standard deviation (S.D.). Statistical evaluation of the data was done by one-way analysis of variance (ANOVA) followed by Bonferroni’s test using In-Stat Graph Pad software. A ‘p’ value less than 0.05 was considered significant with respect to the control group.

RESULTS AND DISCUSSION: Acute inflam-mation induced by carrageenan is the most commonly used method to screen antiinflam-matory agents. Effect of total triterpenes on paw oedema volumes of carrageenan and formalin-induced animals at different time intervals are given in Fig. 1 and 2, respectively. The total triterpenes significantly inhibited the acute inflammation induced by carrageenan Fig. 1 and the chronic inflammation induced by formalin Fig. 2. in the experimental animals in a dose-dependent manner. The standard drug diclofenac (10 mg/kg b. wt. when administered orally, showed 71.74% inhibition of acute and 76.74% inhibition of chronic inflammation. Whereas the total triter-penes, at concentrations 10, 50, and 100 mg/kg b. wt., showed 36.96%, 54.35%, 76.09% inhibition of acute and 41.86%, 62.79% and 79.07% inhibition of chronic inflammation Table 1.

TABLE 1: EFFECT OF TOTAL TRITERPENES ON CARRAGEENAN-INDUCED ACUTE AND FORMALIN-INDUCED CHRONIC PAW OEDEMA IN MICE

Values are Mean ± S.D., n=6, ***P<0.001, **P<0.01, nsP>0.05 with respect to control, TT- total triterpenes.

The development of carrageenan or formalin-induced oedema is a biphasic inflammatory process ^25-27. Reactive oxygen species such as superoxide, hydroxyl radical, and hydrogen peroxide play a vital role in the oedema formation by these agents ^{28 29}. The early phase of this inflammatory process is involved in the production of histamine, leukotrienes, platelet-activating factor, and possibly cyclo-oxygenase products. The delayed phase of the inflammatory response has been linked to neutrophil infiltration, formation of neutrophil-derived free radicals or oxidants such as H₂O₂, O_2, and OH, and the release of other neutrophil-derived mediators ^{30, 31}. Inhibitors of reactive oxygen species reduce the severity of inflammation, and hence, the administration of anti-oxidants can have a protective role in these conditions. Most of the anti-inflammatory drugs act as an antioxidants and scavenge free radicals generated during the inflammatory process. The result of the present study indicates that 100 mg/kg b. wt. total triterpenes possess significantly higher anti-inflammatory activity than standard drug diclofenac in attenuating both acute and chronic inflammation. Earlier studies conducted in our laboratory revealed potential in-vitro and in vivo antioxidant activities of total triterpenes 11, and this antioxidant power might have helped the total triterpenes in scavenging the free radicals produced during the delayed phase of inflammation.

Freund’s complete adjuvants (FCA) are irreplaceable components in the induction of autoimmune diseases and are the most commonly used immune-adjuvants in experimental research ³². Sub-planar injection of Freund’s complete adjuvant in the rat hind paw led to the development of arthritis. Adjuvant arthritis shares many features with human rheumatoid arthritis, and it affects most of the joints and associated tissues ^{33, 34}. Results of the present investigation reveal that total triterpenes of G. lucidum possessed pronounced anti-inflammatory properties in the immunologically mediated inflammatory response induced by injection of Freund’s complete adjuvant in rats. The change in the paw oedema volume during 22 days after inoculation of Freund’s complete adjuvant is given in Fig. 3. Total triterpenes, at concentrations 10, 50, and 100 mg/kg b. wt., when administrated orally, showed 58.73%, 73.02%, and 79.37% inhibition of paw oedema in the arthritis rats Table 2. Standard drug diclofenac (10 mg/kg b. wt.), when administered orally, showed 80.95% inhibition of the paw oedema Table 2. Adjuvant-induced arthritis in rats is a chronic disease that develops into two phases: acute periarticular inflammation followed by a phase of bone involvement ³⁵. In the FCA injected animals, the injected right hind paw showed a biphasic inflammatory response with an immediate acute phase at day post-inoculation followed by a delayed chronic phase that reached peak oedema on day 10 of the FCA inoculation. In the control and vehicle group, there was not much reduction in the paw oedema and observed a continued chronic phase.

In the treated groups, the paw thickness was found to be decreased, showing a reduction in the chronic inflammation phase, with the maximum effect on the 22^nd day. On the 22^nd day, paw volumes of diclofenac and 100 mg/ kg b.wt. total triterpenes treated animals were found to be reverted to the normal levels, indicating their efficiency in curing the inflammation induced by FCA.

Similarly, in the control group, the adjuvant-induced arthritic animals were presented with symptoms such as thickening of hind paws, inflammation on forepaws, redness, and swelling on ears and nose. All these symptoms were markedly reduced in the total triterpenes, and diclofenac treated group Table 2.

FIG. 1: EFFECT OF TOTAL TRITERPENES ON PAW OEDEMA VOLUMES OF CARRAGEENAN-INDUCED ANIMALS Values are Mean ± S.D., n=6; TT- total triterpenes (mg/kg b.wt.)

FIG. 2: EFFECT OF TOTAL TRITERPENES ON PAW OEDEMA VOLUMES OF FORMALIN-INDUCED ANIMALS Values are Mean ± S.D., n=6; TT- total triterpenes (mg/kg b.wt.)

FIG. 3: EFFECT OF TOTAL TRITERPENES ON PAW OEDEMA VOLUMES OF FCA-INDUCED RATS Values are Mean ± SD, n=6; TT- total triterpenes (mg/kg b.wt.).

TABLE 2: EFFECT OF TOTAL TRITERPENES ON PAW THICKNESS AND NUMBER OF SECONDARY LESIONS IN F C A-INDUCED ARTHRITIC RATS

Values are Mean ± SD, n=6. ***P<0.001, ^nsP>0.05 with respect to control, TT- total triterpenes (mg/kg b.wt).

Oxygen-free radicals and H₂O₂ are closely involved in the pathogenesis of rheumatoid arthritis ³⁶. Superoxide anion or other reactive oxygen species (ROS) liberated by phagocytes are recruited to inflammation sites, which is considered the primary cause of tissue damage associated with many chronic inflammatory diseases ^{37, 38}. Superoxide radicals and hydrogen peroxide, which in the presence of traces of iron salts found in synovial fluids, interact to form the highly reactive hydroxyl radical ^{37, 38, 39}. Granulocytes are sharply increased in number in this disease, and they produce large amounts of O₂^- and H₂O₂ during phagocytosis of immune complexes and other materials. The effect of total triterpenes in neutralizing these free radicals in FCA-induced arthritis rats was evaluated by determining the activities of a non-enzyme anti-oxidant GSH, and the enzymatic anti-oxidants SOD, CAT, and GPx. The activity of enzymatic anti-oxidants and glutathione was decreased in the arthritic control rats compared to the normal rats. But the administration of total triterpenes increased the activity of these enzymes in a dose-dependent manner Table 3. The catalase activity in arthritic rats was 71.03 ± 16.06 K/g Hb; while in the 100 mg/kg b.wt of total triterpenes treated group, it was improved to 178.13 ± 21.71 K/g Hb. Similarly, the SOD activity in the control group of animals was 1.95 ± 0.21 U/ml, which was enhanced to 5.74 ± 2.74 U/ml by 100 mg/kg b.wt of total triterpenes treatment. Treatment with the total triterpenes significantly enhanced the glutathione antioxidant system. The activity of GSH in total triterpenes (100 mg/kg b.wt) treated animals was 29.00 ± 14.53nmol/ml of haemolysate. The activity of GPx in blood was also increased to 3.89 ± 0.48 U/ml after treatment with 100 mg/kg b.wt of total triterpenes.

TABLE 3: EFFECT OF TOTAL TRITERPENES ON BIOCHEMICAL PARAMETERS IN BLOOD AND SERUM OF ARTHRITIC RATS

Values are Mean ± SD, n=6. ***P<0.001, **P<0.01, *P<0.05, nsP>0.05 with respect to control, TT- total triterpenes (mg/kg b. wt)

ROS is the key to joint destruction ^{37, 38}. Once generated, these free radicals provoke deleterious effects on various cellular targets, foremost among which are membrane lipids, leading to an extensive process of lipid peroxidation. The increment of lipid peroxide levels in the synovial fluid, serum, and tissue of arthritic animals and patients suffering from arthritis has already been demonstrated ^{39, 40}. Furthermore, the aggravation of arthritis was associated with the enhancement of lipid peroxidation. The effect of total triterpenes on lipid peroxide formation in the serum of the control and treated groups was evaluated, and the results are given in Table 3. The FCA-treated control group showed a substantial increase in the MDA level as compared to the normal groups. But the administration of total triterpenes significantly decreased the serum lipid peroxide level, thereby preventing the joint destruction. Investigations by other researchers also revealed the potential anti-inflammatory action of various extracts of Ganoderma lucidum in animals ^{5, 6, 7, 8, 9}. Both mycelium and fruiting bodies of Ganoderma lucidum were found to be highly beneficial against carrageenan-induced acute and formalin-induced chronic inflammation in mice ^{8, 9}. Moreover, Ganoderma lucidum extracts expressed equivalent efficacy as standard anti-inflammatory drugs in reducing inflammation. Additionally, Ganoderma lucidum extracts did not show any side effects, such as thymic involution or gastropathy, commonly exhibited by steroids or other non-steroidal anti-inflammatory drugs. All these findings support the strong therapeutic potential of Ganoderma lucidum extracts against inflammation associated-diseases.

CONCLUSION: Total triterpenes significantly inhibited inflammation in both carrageens and formalin models, as evident from the decrease in paw oedema. The anti-inflammatory activity of the total triterpenes at specific doses was significantly higher than the standard drug, diclofenac. It also diminished the inflammation and other arthritic symptoms induced by Freund’s complete adjuvant in Wistar rats. The activities of the antioxidant enzymes (SOD, GPx and catalase) and the level of GSH were restored to the normal level in the total triterpenes treated groups. Administration of different doses of triterpenes effectively attenuated the enhanced serum lipid peroxidation due to FCA administration. The results of the present study thus indicate the significant anti-inflammatory and anti-arthritic activity of Ganoderma lucidum total triterpenes.

ACKNOWLEDGEMENT: Nil

CONFLICT OF INTEREST: The authors declare that there are no conflicts of interest in publishing the manuscript. All the authors are associated and contributed to the work, and there are no competing financial interests.

REFERENCES:

1. Singh N, Baby D, Rajguru JP, Patil P, Thakkannavar S and Pujari VB: Inflammation and cancer. Ann Afr Med 2019; 18(3): 121-26.
2. Paquissi FC: The role of inflammation in cardiovascular diseases: the predictive value of neutrophil–lymphocyte ratio as a marker in peripheral arterial disease. Ther Clin Risk Manag 2016; 12: 851-60.
3. Marksa R and Allegrantea JP: Prevalence and impact of arthritis opportunities for prevention. Health Education Journal 2007; 66(1): 3-21.
4. Okamoto H and Kamatani N: A CCR-5 antagonist inhibits the development of adjuvant arthritis in rats. Rheumatology 2006; 45: 230-32.
5. Lacin N, Izol SB, Ipek F and Tuncer MC: Ganoderma lucidum, a promising agent possessing antioxidant and anti-inflammatory effects for treating calvarial defects with graft application in rats.Acta Cir Bras 2019; 25; 34(9): e201900904.
6. Feng X and Wang Y: Anti-inflammatory, anti-nociceptive and sedative-hypnotic activities of lucidone D extracted from Ganoderma lucidum. Cell Mol Biol (Noisy-le-grand) 2019; 65(4): 37-42.
7. Chen YS, Chen QZ, Wang ZJ and Hua C: Anti-Inflammatory and hepa to protective effects of Ganoderma lucidum polysaccharides against carbon tetrachloride-induced liver injury in kunming mice. Pharmacology 2019; 103(3-4): 143-50.
8. Lakshmi B, Ajith TA, Sheena N, Gunapalan N and Janardhanan KK: Antiperoxidative, anti-inflammatory, and anti-mutagenic activities of ethanol extract of the mycelium of Ganoderma lucidum occurring in South India. Teratog Carcinog Mutagen 2003; 23(S1): 85-97.
9. Joseph S, Sabulal B, George V, Antony KR and Janardhanan KK: Antitumor and anti-inflammatory activities of polysaccharides isolated from Ganoderma lucidum. Acta Pharm 2011; 61:335-42.
10. Ho YW, Yeung JSL, Chiu PKY, Tang WM, Lin ZB, Man RYKand Lau CS: Ganoderma lucidum polysaccharide peptide reduced the production of pro inflammatory cytokines in activated rheumatoid synovial fibroblast. Mol Cell Bio Chem 2007; 301:173-79.
11. Smina TP, Mathew J, Janardhanan KK and Devasagayam TPA: Antioxidant activity and toxicity profile of total triterpenes isolated from Ganoderma lucidum (Fr.) P Karst occurring in South India. Env Toxicol Pharm 2011; 32: 438-46.
12. Harborne JB: Phytochemical methods a guide to modern techniques of plant analysis. Chapman and Hall London 1973.
13. Winter CA, Risley EA and Nuss GW: Carrageen an-induced oedema in the hind paw of rat as an assay for anti-inflammatory activity. Proc Soc Exp Biol Ther 1962; 111: 544-47.
14. Demsie DG, Yimer EM, Berhe AH, Altaye BM and Berhe DF: Anti-nociceptive and anti-inflammatory activities of crude root extract and solvent fractions of Cucumis ficifolius in mice model. J Pain Res 2019; 12: 1399-09.
15. Kumar Tand Jain V: Antinociceptive and anti-inflammatory activities of Brideliaretusa methanolic fruit extract in experimental animals. The Scientific World Journal 2014; 2014: 1-12.
16. Jitta SR, Daram P, Gourishetti K, Misra CS, Polu PR, Shah A, Shreedhara CS, Nampoothiri M and Lobo R: Terminalia tomentosa Bark Ameliorates Inflammation and Arthritis in Carrageen an Induced Inflammatory Model and Freund’s Adjuvant-Induced Arthritis Model in Rats. Journal of Toxicology 2019, 2019: 1-11.
17. Snekhalatha U, Anburajan M, Venkatraman B and Menaka M: Evaluation of complete Freund's adjuvant-induced arthritis in a Wistar rat model. Comparison of thermography and his to pathology. Z Rheumatol 2013; 72(4): 375-82.
18. Satoh K: Serum lipid peroxide in cerebro vascular disorders determined by new colorimetric method. Clin Chim Acta 1987; 90: 37-43.
19. Ohkawa H, Ohishi N and Yagi K: Assay for lipid per oxidation in animal tissues by thiobarbituric acid reaction. Anal Bio Chem 197995: 351-58.
20. Drabkin DL and Austin JM: Spectophotometric studies, spectrometric constants for common haemoglobin derivatives in human, dog and rabbit blood. J Biol Chem 1932; 98: 719-33.
21. McCord JM and Fridovich I: Superoxide dismutase enzyme function for erythrocaprein. J Biochem 1969; 244: 6049-56.
22. Aebi M: Catalase estimation In: berg meyer hv editor.methods of enzymatic analysis. New York Verlag Chemie 1974; 673-84.
23. Hafemann DG, Sunde RA and Housestra WG: Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in rat. J Nutrition 1974; 104: 580-84.
24. Maron DM and Ames BN: Revised methods for the Salmonella mutagenicity test. Mut Res 1983; 113: 173-15.
25. J Damas and Liegeois JF: The inflammatory reaction induced by formalin in the rat paw. Naunyn Schmiedebergs Arch Pharma 1999; 359(3): 220-7.
26. Prabu D, Kirubanandan S, Ponnudurai K, Nappinnai M, Jinu AJS and Renganathan S: Anti-inflammatory and analgesic activities of methanol extract of Triphala - a poly herbal formulation. Orient Pharm Exp Med 2008; 8(4): 423-49.
27. Yin Z-Y, Li L, Chu S-S, Sun Q, Ma Z-LandGu X-P: Antinociceptive effects of dehydrocorydaline in mouse models of inflammatory pain involve the opioid receptor and inflammatory cytokines. Sci Rep 2016; 6:27129.
28. Adefegha SA, Leal DBR, de Oliveira JS, Manzoni AG and Bremm JM: Modulation of reactive oxygen species production, apoptosis and cell cycle in pleural exudate cells of carrageenan-induced acute inflammation in rats by rutin. Food Funct 2017; 8(12): 4459-68.
29. Barth CR, Funchal GA, Luft C, de Oliveira JR, Porto BN and Donadio MV: Carrageen an-induced inflammation promotes ROS generation and neutronphil extra cellular trap formation in a mouse model of peritonitis. Eur J Immunol 2016; 46(4): 964-70.
30. Oshishi S, Hayashi I, Hyashi M, Yamaki K and Utsunomiya I: Pharmacological demonstration of inflammatory mediators using experimental inflammatory models: Rat pleurisy induced by carrageen an and phorbolmyristate acetate. Dermatologia 1989; 179: 68-71.
31. Salvemini D, Wang ZQ, Bourdon DM, Stern MK, Currie MG and Manning, PT: Evidence of peroxy nitrate involvement in the carrageenan induced rat paw oedema. Eur J Pharmacol 217-20.
32. Billiau A and Matthys P: Modes of action of Freund's adjuvants in experimental models of autoimmune diseases. J Leukoc Biol 2001; 70(6): 849-60.
33. Billiau A and Matthys P: Collagen-induced arthritis and related animal models: how much of their pathogenesis is auto-immune, how much is auto-inflammatory. Cytokine Growth Factor Rev 2011; 22(5-6): 339-44.
34. Pearson CM: Development of arthritis, periarthritis and periostitis in rats given adjuvants. Proc Soc Exp Biol Med 1956; 91(1): 95-101.
35. Jacobson PB, Morgan SJ and Wilcox DM: A new spin on an old model: in-vivo evaluation of disease progression by magnetic resonance imaging with respect to standard inflammatory parameters and histopathology in the adjuvant arthritic rat. Arthritis Rhe 1999; 42(10): 2060-73.
36. Quinonez-Flores CM, Gonzalez-Chavez SA, Naajera DDR and Pacheco-Tena C: Oxidative stress relevance in the pathogenesis of the rheumatoid arthritis: a systematic review. Bio Med Research International 2016; 1-14.
37. Mittal M, Siddiqui MR, Tran K, Reddy SP and Malik AB: Reactive oxygen species in inflammation and tissue injury. Antioxid Redox Signal 2014; 20(7): 1126-67.
38. Hitchon CA and El-Gabalawy HS:Oxidation in rheumatoid arthritis. Arthritis Res Ther 2004; 6(6):265–278.
39. Tiku ML, Shah R and Allison GT: Evidence linking chondrocyte lipid per oxidation to cartilage matrix protein degradation Possible role in cartilage aging and the pathogenesis of osteoarthritis. J Biol Chem 2000; 275: 20069-76.
40. Dai L, Lamb DJ, Leake DS, Kus ML, Jones HW, Morris CJ and Winyard PG: Evidence for oxidised low density lipoprotein in synovial fluid from rheumatoid arthritis patients. Free Radic Res 2000; 32: 479-86.
    

    ---

    How to cite this article:

    Smina TP, Mathew J, Rony KA and Janardhanan KK: Ganoderma lucidum total triterpenes alleviate inflammation and modulate antioxidant status in freund’s complete adjuvant-induced arthritic rats. Int J Pharm Sci & Res 2021; 12(6): 3490-97. doi: 10.13040/IJPSR. 0975-8232.12(6).3490-97.

    ---

    All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

    ---