Evaluation of Anti-arthritic activity of Methanolic extract of Barleria prionitis on CFA induced rats

 

G. Sivakumar¹*, G. Arihara Sivakumar², Rebecca²

¹Assistant Professor, Department of Pharmacology, KMCH College of Pharmacy, Kalapatti Road,

Coimbatore - 641048 Tamilnadu, India

²Department of Pharmacology, KMCH College of Pharmacy, Coimbatore-48

 

ABSTRACT:

Objective: To investigate the anti-arthritic activity of methanolic extract of Baleria prionitis (MEBP) in complete Freund’s (CFA) induced rats. Materials and Methods: Arthritis was induced in rats following a subplantar injection of Freund’s complete adjuvant (0.1ml). Rats were divided into six groups of six animals each. Group-I and II were control injected with normal saline and CFA (0.1ml), respectively. Group-III, IV, V were received Leflunomide (LFD) (10 mg/kg, p.o.), MEBP (200 mg/kg, p.o.) and MEBP (400 mg/kg, p.o.), respectively, followed by the induction of arthritis with CFA (0.1ml) single dose on 0^th day for 28 days. Measurement of joint diameter was again carried out on 1^st, 7^th, 14^th, 21^st, and 28^th days. On the 28^st day of the study, blood sample was collected for estimation of serum arthritic parameters such as C-reactive protein (CRP) and Rheumatoid factor (RF). Radiological analysis and haemoglobin estimation were also carried out at the end of and 28^th day. Results: There was a significant inhibition (P<0.01) in paw edema at a different time scale with doses of MEBP (200 mg/kg and 400 mg/kg). MEBP also demonstrated dose dependent anti-arthritic activity as well as superior activity over standard drug LFD on both doses of MEBP on all observations days from 7^th, 14^th, 21^st, and 28^th days. Serum biochemical parameters such a RF and CRP were found to be significantly reduced as compare to control group and close to the LFD treated group. In both LFD and MEBP treated groups showed no visible sign of bone and joint deformation as well as suppression of inflammation and subsequent arthritic joint development was clearly visible by radiological analysis. Hemoglobin protective effect of MEBP was also obsrved as compare to LFD treated group. Conclusion: This study demonstrates that MEBP possesses potent anti-arthritic activity and protection from hematological toxicity compare to standard drug.

 

 

 

INTRODUCTION:

Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology and is mainly characterized by the progressive erosion of cartilage leading to chronic polyarthritis and joint distortion. [1] RA predominantly occurs in females. The prevalence of RA is around 1% worldwide, with women suffering 3-5 times more than men. RA is very common disease in India affecting elderly ladies. The Indian prevalence rate (0.9%) almost equals to the world prevalence rate. The variation in the level of sex hormones of women (estrogen and progesterone, which regulate the inflammatory process) is the main cause of development of RA among them. Globally, 1.3 million adults suffer from RA. [2] Treatment of RA mainly includes non-steroidal anti-inflammatory drugs, glucocorticoids, Disease Modifying Anti-Rheumatic Drugs (DMARD) [3,4,5]. Recent report suggests the p38 Mitogen Activated Protein Kinase (MAPK) plays an important role in the production of proinflammatory cytokines, making it an attractive target for the treatment of Rheumatoid Arthritis and various inflammatory diseases. [6]

 

In complementary and alternative medicines such as Ayurveda (herbs) and acupuncture are most commonly used for the treatment of many systemic disorders. At present, electromagnetic fields (EMFs), [7] Nordic walking (with pole) and Isokinetic Training [8] and Electromyography biofeedback training [9] have shown positive therapeutic effects in this disorder.  Chopra et al., reported around 68% patients with RA has sought relief using alternative system of medicine and demonstrated the clinical efficacy of herbal formulation for the treatment of osteoarthritis of the knees. [10,11, 12] Recent reports suggest LFD belongs to the class of DMARD was also proven to effective and well tolerated in the treatment of rheumatoid arthritis (RA). Its beneficial effects showed in high American College of Rheumatology (ACR) response rates and comprised retardation of radiographic progression as well as improvement of physical function and health-related quality of life. [13] However LFD can cause abnormal liver function tests [14] and decreased blood cell (or) platelet counts.

 

Barleria prionitis Linn. belongs to the family Acanthaceae (BPA) which is a prickly shrub 1.5 m high with simple opposite decussate leaves. The flower is yellow and sessile. The plant is cultivated as hedge plant and is distributed throughout the warmer parts of India. [15,16] The major phytochemical constituents found are acetyl-barlerin and barlerin, [17,18] and other chemical constituents present are 6-O-acetylshanzhiside methyl ester, [19] 6-O-cis-p-coumaroyl-8-O-acetylshanzhiside methyl ester its trans-isomers, shanzhiside methyl ester α-amyrin, verbascoside, [20] β-sitosterol, [21] and stigmasterol-3-O-D-glucoside. The plant is known to have anti-bacterial [22] anti-inflammatory, analgesic, [23] anti-oxidant, [24] expectorant, diuretic, anti-rheumatic, and anti-diabetic properties. [25] The plant extract is also known to possess hepatoprotective activity, [26]   in respiratory infections and tuberculosis. Current literature study revealed that the aerial parts of Barleria prionitis has not been studied for its anti-arthirtic activity, hence the aim of the present study is to evaluate the anti-arthritic activity of methnolic extract of Barleria prionitis Linn. on CFA induced arthritic rats with LFD as standard drug.

 

MATERIALS & METHODS:

Plant Material:

At the flowering stage, aerial parts of the plant Barleria prionitis were collected from Khammam, Andhra Pradesh, India during the month of December and authenticated by Botanical Survey of India (BSI), Southern Circle, Coimbatore, Tamilnadu. Soon after collection, the aerial parts were cleaned, dried in shade and crushed to a coarse powder, stored in an air tight plastic container, until further use.

 

Preparation of Extract of Barleria prionitis:

The 300g of the dried coarse powder of Barleria prionitis aerial parts was first treated with petroleum ether, the defatted powder was extracted with methanol using soxhlet apparatus and the percentage yield was found to be 2.8 % w/w. The dried extracted material was stored at −20°C until use.

 

Animals:

Female Albino Wistar rats of 6-8 weeks old and 160-180 g body weight were offered by KMCH College of pharmacy, Coimbatore. All rats were kept at room temperature (25◦C ± 2◦C) and allowed to accommodate in standard conditions at 12-hr light and 12-hr dark cycle in the animal house. Animals were fed with standard pellet diet and water ad libitum freely.

 

Adjuant-Induced Arthritis:

Rats were divided into six groups of six animals each. Group-I and II were control injected with normal saline and CFA (0.1ml), respectively. On day 1, baseline measurement of the paw diameter was carried out. Digital Vernier Caliper was used for the measurement of the joint size. The severity of arthritis was recorded by scoring system. [27] Arthritis was induced in rats following a subplantar injection of Freund’s complete adjuvant (0.1ml). It was done 30 minutes after the administration of vehicle/drugs to the respective groups of animals. This was followed by daily administration of vehicle/drugs to the respective groups for 28 days. Measurement of joint diameter was again carried out on 1^st, 7^th, 14^th, 21^st, and 28^th days.

 

Experimental design for evaluating anti-arthritic activity:

1.     Group-1 Vehicle control

2.     Group-II Negative control treated with CFA 0.1 ml on the left hind paw

3.     Group-III Positive control treated with CFA+ LFD 10mg/kg, p.o.

4.     Group-IV MEBP treated with CFA+ low dose of MEBP 200mg/kg, p.o.

5.     Group-V MEBP treated with CFA+ high dose of MEBP 400mg/kg, p.o.

Evaluation of Arthritis:

Paw Diameter:

Paw thickness was measured by compressing the joint by rotating the screw of micrometre screw gauge till the pain elicited as indicated by squeaking or leg withdrawal. The distance moved by the screw gauge was recorded.

 

Serum Biochemical Parameters - C-Reactive protein detection [28]:

The CRP-HS is intended for the quantitative determination of CRP by latex principle enhanced Immune Turbidimetric Assay (ITA). Latex particles coated with antibody specific to human CRP aggregate in the presence of CRP from the sample forming immune complexes. The immune complexes cause a light scattering which is proportional to the concentration of CRP in the serum. The light scattering is measured by reading turbidity (absorbance) at 570 nm. The CRP concentration is determined from a calibration curve developed from CRP standards of known concentration. [29]

 

Rheumatoid factor detection [30]:

Rheumatoid factor (RF) is an anti-antibody, which in-vitro, is detected by its ability to agglutinate latex particles (or red blood cells) coated with human IgG. RF will attach to the IgG coating the latex particles. Agglutination of the latex particles is a positive result indicating the presence of RF. Turbidimetric immunoassay is used for the quantitative detection of rheumatoid factor of IgM class. [30]

 

Hematological Analysis - Estimation of haemoglobin:

The Haemoglobin is located with a reagent containing potassium ferrocyanide, potassium cyanide and potassium dihydrogen phosphate. The ferrocyanide forms methhemoglobin which is converted to cyanomethhemoglobin by the cyanide. [31] 0.2µl of blood was added to 5ml of Drabkin’s reagent. It was allowed to stand for at least 5minutes after the addition of the reagent. The solution was read against an absorbance of 540nm.

 

Radiological Study [32]:

The radiological analyses of hind paw of All groups were performed in order to compare the synovitis, pannus formation, focal cartilage or bone erosion, joint deformation, suppression of inflammation and subsequent arthritic joint development to assess the RA protective effect of LFD and MEBP treated groups.

RESULTS:

Inhibition of adjuvant arthritis by Barleria prionitis:

The anti-arthritic action of MEBP following the subplantar injection of CFA has been tabulated [Table-1].  There has been gradual increase in joint diameter following 0^th day after administration of CFA, which was maximum on day 7. After that, there was a progressive decrease in joint diameter in all the groups except the vehicle-treated where it was increased up to some extent after the 14^th day. Treatment with LFD, MEBP (200 mg/kg, p.o.) and MEBP (400 mg/kg, p.o.), resulted in a remarkable decline in joint diameter and the decline was statistically significant in comparison to vehicle-treated group depicted in [Figure-1].

 

Effect of Barleria prionitis on Biochemical and Hemoglobin Parameters in FDA induced arthritic rats:

The anti-arthritic effect of MEBP was also evidenced by biochemical parameters, tabulated on [Table-2].  There was a significant increased levels of biochemical parameters observed in CFA treated arthritis group, whereas statistically significant reduction in both Rhematoid Factor and C-Reactive protein factors was found on LFD, MEBP (200 mg/kg, p.o.) and MEBP (400 mg/kg, p.o) treated groups depicted on [Figure- 2 & 3], respectively. The Hemoglobin protection effect of MEBP was shown on [Table-3].  In both MEBP (200 mg/kg, p.o.) and MEBP (400 mg/kg, p.o) treated groups Hemoglobin levels were not reduced, compare to LFD drug treated group at a statistically significant level, depicted on [Figure-4].

 

Radiological Assessment:

The radiological analyses of hind paw of Control, Arthritic Control, LFD and MEBP (400 mg/kg, p.o.) groups were performed and compared, depicted on [Figure- 5,6,7,8,9], respectively. In normal control group, no synovitis, pannus formation, focal cartilage or bone erosion was seen in ankle joints, while arthritic control group showed cell infiltrate, synovial hyperplasia, periarthritis and focal bone erosion. In both LFD and MEBP (400 mg/kg, p.o.) treated groups showed no visible sign of bone and joint deformation, as well as suppression of inflammation and subsequent arthritic joint development was clearly visible.

 

Statistical Analysis:

The data obtained after studying the various effect of Barleria prionitis using the appropriate rat model were revealed in the form of mean ± standard error of the mean. The method that was adopted to find the difference in significance from the statistical point of view was one-way ANOVA followed by Dunnett's multiple comparison. The value that was considered significant from the statistical point of view was <0.05.

Table-1: Effect of MEBP on CFA induced arthritis paw diameter

Groups	Days
	0 (mm)	7 (mm)	14 (mm)	21(mm)	28 (mm)
Normal Control	6.83 ± 0.12	6.89 ± 0.32	7.11 ± 0.60	7.02 ± 0.54	7.12 ± 0.80
Arthritic Control	6.86 ± 0.20a	14.52 ± 0.55a	15.14 ± 0.68a	15.11 ± 0.47a	14.80 ± 0.75a
CFA + LFN (10mg/kg)	6.73 ± 0.17	10.57 ± 0.61*	9.30 ± 0.69**	8.57 ± 0.73***	7.30 ± 0.71
CFA + MEBP (200mg/kg)	6.90 ± 0.19	12.28 ± 0.65ns	12.09 ± 0.98*	11.27 ± 1.24*	10.32 ± 1.04**
CFA + MEBP (400mg/kg)	6.89 ± 0.13	12.13 ± 0.53ns	11.47 ± 0.69*	9.82 ± 0.91**	8.89 ± 0.90***

 

Statistical comparison: Each group (n=6), each value represents Mean ± SEM. One way Annova followed by Dunnett’s test was performed. ^aP<0.001 denotes comparison of Arthritic control with vehicle control and ns- non significant ^*P<0.05, ^**P<0.01, and ^***P<0.001 denotes comparison of all groups with Arthritic control

 

Table-2: Effect of MEBP on biochemical and Hematological parameters in CFA treted rats

Serum Biochemical Parameters			Hematological Parameter
Groups	Rheumatoid Factor (IU/L)	C-Reactive Protein (mg/dl)	Haemoglobin (mg/dl)
Normal Control	15.43 ± 0.9018	3.33 ± 0.6429	13.5±0.35
Arthritic Control	52.95 ± 5.303a	16.96 ± 2.411a	10±0.22
CFA + LFN (10mg/kg)	18.37 ± 2.044**	5.95 ± 0.12**	11.4±0.55*
CFA + MEBP (200mg/kg)	6.90 ± 0.19	12.28 ± 0.65ns	12.6±0.50**
CFA + MEBP (400mg/kg)	6.89 ± 0.13	12.13 ±0.53ns	13.1±0.37***

 

Statistical comparison: Each group (n=6), each value represents Mean ± SEM. One way Annova followed by Dunnett’s test was performed. ^aP<0.001 denotes comparison of Arthritic control with vehicle control and ns- non significant ^*P<0.05, ^**P<0.01, and denotes comparison of all groups with Arthritic control

 

 

Figure-1

Effect of MEBP extract on joint swelling in CFA induced arthritis in rats

All values are mean ± standard error

 

Figure-2

Effect of MEBP extract on Rheumatoid Factor in CFA arthritis in rats

All values are mean ± standard error

 

Figure-3

Effect of MEBP extract on C-Reactive protein in CFA induced arthritis in rats

All values are mean ± standard error

 

Figure-4

Hemoglobin Protectove effect of MEBP extract on CFA induced arthritis in rats

All values are mean ± standard error

Radiological Analysis of hind paw

Figure-5: Normal control.

 

Figure-6: Arthritic Control (CFA)

 

Figure-7: Arthritic Control (CFA)

 

Figure-8: positive control (CFA + LFD 10 mg/kg)

 

Figure-9: (CFA + MEBP 400 mg/kg)

 

DISCUSSION:

Treatment of RA mainly includes non-steroidal anti-inflammatory drugs, glucocorticoids, and DMARD, however important long term serious ADR such as decreased blood cells and abnormal liver functions associated with DMARD demands, complementary and alternative medicines such as Ayurveda and acupuncture. Recent report suggests that around 68% patients with RA sought relief using alternative system of medicine and demonstrated the clinical efficacy of herbal formulation for the treatment of osteoarthritis of the knees. Hence the present work is aimed to evaluate the anti-arthritic activity of methnolic extract of Barleria prionitis Linn. on CFA induced arthritic rats with LFD as standard drug.

 

CFA-induced arthritis is a very suitable model for testing anti-arthritic activity as it has a very high degree of validity and has many similarities with human arthritis. [33, 34] Recent reports suggest LFD belongs to the class of DMARD was also proven to effective and well tolerated in the treatment of rheumatoid arthritis (RA). Based on the literature review LFD, [35] MEBP (low and high) doses were selected 10 mg/kg, 200 mg/kg and 400 mg/kg, respectively.

 

The statistically significant positive results were observed in CFA-induced arthritis is a very suitable model with human arthritis. The same degree of positive result were also observed in both biochemical and radiological analysis, which reinforces the potent anti-arthritic activity of MEBP by multiple inhibitory mechanism such as anti-oxidant, anti-inflammatory and immuno-modulatory activity related to pathology of development of RA with dose related increase activity. In addition to that MEBP also has haemoglobin protective effect as compared to LFD, hence MEBP would be a potential new drug target for anti-arthritic activity.

 

The important phytochemical principles to be present in MEBP are flavonoids, irido- glycosides, phenolic compounds and steroids. The anti-arthritic activity of MEBP may most likely due flavonoids as it was reported in most of the animal experiment to possess anti-inflammatory, anti-oxidant activity and inhibiting phospholipase A₂ as it is more relevant to pathology of RA, however recent reports reveals iridoid-glycoside also possess the same activities. Due to the presence of the multiple phytochemical presents in MEBP, it is difficult to narrow down a particular phyto-chemical principle responsible for anti-arthritic activity, therefore isolated phytochemical constituents anti-arthritic activity would be needed to conclude the precise phytochemical responsible for the activity, which would be the future direction to explore and invent new drug development  to treat RA more effectively with  minimal adverse drug reactions than the existing drugs.

 

FINANCIAL SUPPORT AND SPONSORSHIP:

Nil.

 

CONFLICTS OF INTEREST:

There is no conflicts of interest.

 

ACKNOWLEDGEMENTS:

I would like to thank Dr. G. Arihara Sivakumar, Head of the Department of Pharmacology and Dr. A. Rajasekaran, Principal KMCH College of Pharmacy for the constant encouragement for preparing this research article. I feel grateful to the Chairman and Trustee Madam of KMCH College of Pharmacy for providing valuable infrastructure facility to perform this research work.

 

REFERENCES:

1.      McInnes IB, Schett G. The pathogenesis of rheumatoid arthritis. N Engl J Med 2011; 365:2205-19.

2.      Shikha Srivastava, Shatish Patel, S.J. Daharwal, Deependra Singh, Manju Singh. Rheumatoid Arthritis: An Autoimmune Disease Prevalent in Females. Research J. Pharm. and Tech. 9(2): Feb., 2016; Page 170-172.

3.      Sarumathy S., CH Haripriya, Muddala Vara Prasanna Rao, Gayathri K., CH Bala Sudhakar, Divya M., Shanmugasundaram P. Clinical Comparison of the Efficacy and Safety of Intra-Articular Injections of Sodium Hyaluronate and MethylPrednisolone in the Treatment of Osteoarthritis of Knee. Research J. Pharm. and Tech. 8(11): Nov., 2015; Page 1526-1528.

4.      Gayathri Devi Kumaresan, Dhanraj M. Efficacy of Cox-2 inhibitors in the Clinical Management of TMJ Arthritis: A Review. Research J. Pharm. and Tech 2017; 10(12): 4439-4441

5.      B. Premkumar Antioxidant Defense and Disease activity in Rheumatoid Arthritis. Research J. Pharm. and Tech 2018; 11(5):1810-1814.

6.      Radha Mahendran, Suganya Jeyabasker, Astral Francis, Sharanya Manoharan. Insights into the Identification of p38-alpha Mitogen activated Protein Kinase against Pyridazinopyridinone Derivatives in the Treatment of Rheumatoid Arthritis. Research J. Pharm. and Tech. 2017; 10(9): 2875-2879.

7.      Ali Yadollahpour, Samaneh Rashidi. Electromagnetic fields for the treatment of osteoarthritis: A review of potential clinical applications. Research J. Pharm. and Tech. 2017; 10(2): 641-644.

8.      M. Subramoniam, T. Jerry Alexander. A Novel Method for Early Diagnosis of Arthritis from Digital X-Ray Images. Research J. Pharm. and Tech. 2017; 10(6): 1611-1614.

9.      Naresh Bhaskar Raj, Soumendra Saha, Amran Ahmed Shokri, Srilekha Saha, Hazliza Razali, Nur Yanti Hariana Othman, Mahadeva Rao US. Effect of Isokinetic Training on Quadriceps Muscle Strength in Osteoarthritis of Knee. Research J. Pharm. and Tech 2018; 11(6): 2517-2524.

10.    Mukherjee PK, Venkatesh P, Ponnusankar S. Ethnopharmacology and integrative medicine - Let the history tell the future. J Ayurveda Integr Med. 2010; 1:100–9.

11.    Chopra A, Lavin P, Patwardhan B, Chitre D. A 32-week randomized, placebo-controlled clinical evaluation of RA-11, an Ayurvedic drug, on osteoarthritis of the knees. J Clin Rheumatol. 2004; 10:236–45.

12.    Shyama S. Kumar, Divya Bhosle, Akshay Janghel, Shraddha Deo, Parijeeta Raut, Chetan Verma, Mukta Agrawal, Nisha Amit, Mukesh Sharma, Tapan Giri, D. K. Tripathi1, Ajazuddin, Amit Alexander. Indian Medicinal Plants Used for Treatment of Rheumatoid Arthritis. Research J. Pharm. and Tech. 8(5): May, 2015; Page 597-610.

13.    Sharp JT, Strand V, Leung H, et al. Treatment with leflunomide slows radiographic progression of rheumatoid arthritis: results from three randomized controlled trials of leflunomide in patients with active rheumatoid arthritis. Leflunomide Rheumatoid Arthritis Investigators Group. Arthritis Rheum. 2000; 43:495–505. doi: 10.1002/1529-0131(200003)43:3<495:AID-ANR4>3.0.CO;2-U.

14.    Salmi Abdul Razak, Farida Islahudin, Ahmad Fuad Shamsuddin, Nor Shuhaila Shahril. A study on Leflunomide-induced liver injury in Rheumatoid Arthritis Patients. Research J. Pharm. and Tech. 6(5): May 2013; Page 556-561

15.    Gupta AK, Tandon N, Sharma M. Quality Standards of Indian Medicinal Plants. 1st ed. Vol. 4. New Delhi: Indian Council of Medical Research; 2006. p. 36.

16.    Umesh Dhaked, Gaurav Nama, Devendra P. Singh, Amit K. Mishra, Nitin Kumar. Pharmacognostical and Pharmacological Profile of Barleria prionitis Root. Research J. Pharmacognosy and Phytochemistry 2011; 3(3): 108-111.

17.    Taneja SC, Tiwari HP. Structures of two new iridoids from Barleria prionitis Linn. Tetrahedron Lett. 1975; 24:1995–8.

18.    Damtoft S, Jensen SR, Nielsen BJ. Structural revision of barlerin and acetyl barlerin. Tetrahedron Lett. 1982; 23:4155–6.

19.    El-Emary NA, Makboul MA, Abdel-Hafiz MA, Ahmed AS. Phytochemical study of Barleria cristata L. and Barleria prionitis L. cultivated in Egypt. Bull Pharm Sci Assist Univ. 1990; 13:65–72.

20.    Chen JL, Blanc P, Stoddart CA, Bogan M, Rozhon EJ, Parkinson N, et al. New iridoids from the medicinal plant Barleria prionitis with potent activity against respiratory syncytial virus. J Nat Prod. 1998; 61:1295–7.

21.    Moitra SK, Ganguly AN, Chakravarti NN, Adhya RN. Chemical investigation of Barleria prionitis Linn. Bull Calcutta Sch Trop Med. 1970; 18:7.

22.    Swati Paul, Dibyajyoti Saha. Comparative Study of the Efficacy of Barleria prionitis Leaf Extracts against Bacteria. Asian J. Pharm. Res. 2(3): July-Sept. 2012; Page 107-110.

23.    Sunil. K. Jaiswal, Mukesh. k. Dubey, Sanjeeb Das, Arti. R. Verma, M. Vijayakumar and Chandana.v. Rao. Evaluation of flower of Barleria prionitis for Anti-inflammatory and Anti-nociceptive activity. International Journal of Pharma and Bio Sciences 2010; V1 (2).

24.    Chavan Chetan, Mulik Suraj, Chavan Maheshwri, Adnaik Rahul, Patil Priyanka. Screening of antioxidant activity and phenolic content of whole plant of Barleria prionitis L. International Journal of Research in Ayurveda and Pharmacy 2011; 2(4): 1313-1319.

25.    Pullaiah T, Chandrasekhar Naidu K. Antidiabetic Plants in India and Herbal Based Antidiabetic Research. New Delhi: Regency Publications; 2003. p. 99.

26.    Singh B, Chandan BK, Prabhakar A, Taneja SC, Singh J, Qazi GN, et al. Chemistry and hepatoprotective activity of an active fraction from Barleria prionitis linn. In experimental animals. Phytother Res. 2005; 19:391–404.

27.    Laima L, Eiva B, Ruta B, Dalia V, Elvyra R, Vytautas A. Antiarthritic and hepatoprotective effect of derinat on adjuvant arthritis in rats. Acta Med Lituanica 2006; 13:236-44.9.

28.    Michael Torzewski, Ahmed Bilal Waqar, and Jianglin Fan, “Animal Models of C-Reactive Protein,” Mediators of Inflammation, vol. 2014, Article ID 683598, 7 pages, 2014.

29.    Borque L, Bellod L, Rus A, Seco ML, Galisteo-Gonzalez F. Development and validation of an automated and ultrasensitive immunoturbidimetric assay for C-reactive protein [Technical Brief]. Clin Chem 2000; 46:1839-1842.

30.    Aletaha, D., Neogi, T., Silman, A. J., Funovits, J., Felson, D. T., Bingham, C. O., Birnbaum, N. S., Burmester, G. R., Bykerk, V. P., Cohen, M. D. et al. (2010). 2010 Rheumatoid arthritis classification criteria: an American college of rheumatology/European league against rheumatism collaborative initiative. Arthritis. Rheum. 62, 2569-2581.doi:10.1002/art.27584

31.    Satarupa Roy and P.K. Suresh. Estimation of Antioxidant Enzyme, Catalase Activity from Erythrocytes of Healthy Individuals. International Journal of Recent Biotechnology. 2013, 1 (2): 17-23

32.    Kumar VL, Roy S. Protective effect of latex of Calotropis procera in Freund’s complete adjuvant induced monoarthritis. Phytother Res. 2009; 23:1–5.

33.    Newbould BB. Chemotherapy of arthritis induced in rats by mycobacterial adjuvant. Br J Pharmacol Chemother 1963; 21:127‑36

34.    Kumar R, Gupta YK, Singh S, Arunraja S. Picrorhiza kurroa inhibits experimental arthritis through inhibition of pro-inflammatory cytokines, angiogenesis and MMPs. Phytother Res 2016; 30:112-9. 

35.    Thoss K, Henzgen S, Petrow PK, Katenkamp D, Brauer R. Immunomodulation of rat antigen-induced arthritis by leflunomide alone and in combination with cyclosporin A. Inflamm Res. 1996 Feb;45(2):103-7.

 

 

 

Received on 04.06.2019            Accepted on 10.07.2019           

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