Skip to main content

EVALUATION OF ANTIINFLAMMATORY EFFECT OF Capparis Aphylla Roth. USING EXPERIMENTAL ANIMAL MODELS

Pharma courses

GPAT courses

Apollo Pharmacy hiring

pharma courses

pharma courses

Apollo Pharmacy hiring

ABOUT AUTHORS:
Kanzaria S. H. ,  Patel D. V., Patel K.V., Gandhi T. R.
Anand Pharmacy College.
Anand, Gujarat
*sainikak@yahoo.in

ABSTRACT:
Objective: To evaluate anti-inflammatory effect of Capparis aphylla Roth. (CA) using experimental animal models. Material and method: Female Wistar rats (200 to 250gm) were randomly allocated to six groups (n=6). Except group-I (normal control), arthritis was induced in animals of all groups by injection of 0.2 ml Complete Freund’s Adjuvant (6mg/ml) on day one. Additionally Group III (Std) and Group IV–VI (CA-1, CA-2 and CA-3) received Indomethacin (100mg/kg), CA (300 mg/kg ; 240 mg/kg and 190 mg/kg) from day 1 to day 21 orally. Paw volumes of both sides (measured by plethysmography) was recorded  on day 0, 7, 14, 21. On 7th, 14th, 21stdays the severity of the secondary lesions was evaluated by measuring body weight, arthritic index, Erythrocyte Sedimentation Rate(ESR), Rheumatoid factor(RF), C- Reactive Protein(CRP), Albumin/Globulin(A/G) ratio, X-ray, histopathology of synovial joints. Result: Capparis Aphylla Roth. significantly prevented the freund’s adjuvant induced changes in body weight, serum A/G ratio , arthritic Index, paw edema, ESR, RF, CRP. the results were comparable with the standard drug indomethacin. Conclusion: Antiinflammatory activity of Capparis aphylla Roth. can beattributed to COX-II inhibition leading to decrease in inflammatory mediator.

[adsense:336x280:8701650588]

REFERENCE ID: PHARMATUTOR-ART-1774

INTRODUCTION
Rheumatoid arthritis (RA) is a common chronic inflammatory autoimmune disease of unknown origin. It affects ∼1% of the population and causes the irreversible functional and anatomical joint damage. The have key factors of inflammation and destruction in the rheumatoid joint namely intracellular signaling and proliferation, adhesion, infammation, matrix degradation and angiogenesis that result in a persisting vicious circle. In addition to proinflammatory cytokines, acute phase response (like CRP), abnormal immunoglobulin levels and high rheumatoid factors (RFs) are also considered to be a modulating event during infammatory and immunogenic reaction. . [CHEMICO BIOLOGICAL INTERATION]

Bone loss is a common feature of various inflammatory arthritis Bone erosions results from the activation of an inflammatory response that increases the number and activity of osteoblasts. The complete freund”s adjuvant induced arthritis model represents a systemic inflammatory disease with bone and cartilage changes similar to those observed in RA. The common pathological features of adjuvant arthritis in rats and RA inhuman are joint swelling associated with cellular and pannus invasion of the joint space and bone resorption. Strong bone loss after intense arthritis is induced when adjuvant is injected into the foot pad. The major site of irreversible tissue damage originates from synovium lining  the joint capsule with cartilage and bone, which is often termed the “pannus” and this  is a characteristic feature of RA. [IJPD, CURATIVE EFFECT]

Capparis aphylla Roth.  is commonly known as Caper berry,  Karira, Kerada, Karer, Kair ,  belonging to the family Capparidaceae. Capparis aphylla Roth. has been used traditionally as an anti-inflammatory agent for enlarged cervical glands, sciatica, rheumatoid arthritis, swellings, skin eruptions, ringworm. [C.P. KHAR] Secondly Capparis aphylla Roth. root bark contains spermidine alkaloids which is known for its use in inflammations, asthma and gout.[C.P. KHAR] The different parts of C.aphylla  are  considered to  be  analgesic, diaphoretic, alexeteric, laxative, antihelminthes,  antibacterial, antifungal, antiviral,  and  good in cough,  asthma, ulcers, boils, vomiting, piles and all inflammations.[ANTIHYPERGLYCEMIC, ANTIOXIDENT]

Consequently, the present study has been undertaken to illustrate the beneficial outcome of the drug capparis aphylla Roth.in adjuvant induced arthritic rat model.

[adsense:336x280:8701650588]

MATERIALS & METHODS.

·        Preparation of extract of Capparis aphylla Roth.
3.2 kg root bark of capparis deciduas were percolated three times with ethyl alcohol at room temperature for 2-3 weeks. The solvent from the combined yellowish brown percolate was removed under reduced pressure. 38 g of dark brown viscous residue was taken up in 1N hydrochloric acid and taken out exhudatively with ethyl acetate to remove neutral substances. The aqeous acidic solution was basified with ammonia, yielding yellow precipitates. These yellow precipitates was then extracted with chloroform which on evaporation gave the alkaloid concentrate. The column chromatography of the chloroform extract was carried out in neutral alumina oxide and eluted with a mixture of ethyl acetate:methanol(4:1) and thus isolated the yellow coloured . The yellow compound so obtained contained traces of impurities.

The dragendroffs reagent, methanol, alumina oxide, ethyl acetate, Na azide, Na citrate, succnic acid, bromocresol, brij-95, formaline, formic acid, freund’s adjuvant, carrageenan were obtained from Astron chemical, ahmedabad,Gujarat, India. The total protein kit, RF latex kit, CRP kit were obtained from Coral clinical system, Gujarat,India.

* Animals
Female Wistar albino rats weighing 200–250 g were used. The animals were fed ad libitum and housed in a room with a controlled ambient temperature (22 ±2 °C), humidity (50% ± 10%), and a 12-h light/dark cycle. Animals were acclimated to the housing conditions and handled for 3–4 days before experiments. All experimental procedures were conducted according to the OECD  Guidelines for the Care and Use of Laboratory Animals . The experimental protocols were approved by the Ethical  Committee on Animal Care.

* Preparation of drug solution:
Accurately weighed quantity of dried extract was suspended in distilled water  to prepare the appropriate stock solution of the drug i.e. 190mg/kg, 240mg/kg, 300mg/kg. Indomethacin 100mg/kg was also suspended in distilled water. The doses were administered by selecting theappropriate concentration of the stock solution by orally.

* Animal Grouping
Rats were divided into 6 equal groups (6 rats for each). Group 1 Normal Control group: rats received pure drinking water and regular food ; Group 2 Model  Control group: Freund’s adjuvant; Group 3 Standard group : CFA+ Indomethacin 100mg/kg; Group 4 test group: CFA+MECA 190mg/kg; Group 5 test group: CFA+ MECA 240mg/kg; Group 6 test dose: CFA+ MECA 300mg/kg. Animals were treated with drugs for 21 days.

* Induction of Complete Freund’s Adjuvant Arthritis

- Experimental procedure [Schorlemmer HU,et.al, 1999, 474.]
Female Wistar rats (180 to 250 g ) was be  randomly allocated to seven groups containing six animals each. Animals in Group I(normal control) received distilled water for the entire 21 day study period. Arthritis was be induced in animals of Group III- VII on day one  by injection of 0.2 ml complete Freund’s adjuvant (It consisted of 6 mg Mycobacterium butyricum  being suspended in heavy paraffin oil  to give a concentration of 6 mg/ml) in sub plantar region of the left hind paw. Group III(Std),Group IV(test drug 190 mg/kg), Group V(test drug 240 mg/kg), Group VI (test drug 300mg/kg ). Paw volumes of both sides and body weight was be recorded on the day of injection, whereby paw volume was be measured by plethysmography. On day 7,14,21 the volume of the injected paw was be  measured indicating the primary lesion and the influence of therapeutic agents on this phase. The severity of the induced adjuvant disease was be followed by measurement of the non injected paw (secondary lesions) with a plethysmometer. At the end of study ,on 21 days following parameters was be evaluated.

1.      Body weight[Lin, H-S Hu C-Y,2007, 150, 862–872]
Body weight of each animal was be measured on the day of CFA administration, and later, on 7th, 14th and 21st days. The mean percentage reduction in body weights with respect to that on day of CFA administration was be calculated for each drug treated group and compared with that of disease control group.

2.       Arthritic Index [Arthur Yin Fana, 121,2009 366–371.
On day 21, the severity of the secondary lesions was be evaluated visually and graded all to    following scheme.:

ORGANS

INDICATIONS

SCORING

EARS

Absence of nodules& redness

0


Presence of nodules& redness

1

NOSE

No swelling of connective tissue

0


Intensive swelling of connective tissue

1

TAIL

Absence of nodules

0


Presence of nodules

1

FORE   PAWS

Absence of inflammation

0


Inflammation of at least one jointq

1

HIND PAWS

Absence of inflammation

0


Slight inflammation

1


Moderate inflammation

2


Marked inflammation

3

arthritic score

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

An arthritic index for each animal was be calculated as the sum of these scores. The average scores for each group of drug treated animals was be compared with that of disease control animals

3.       Paw edema[Magari Katsue, 2003 139, 927–934.]
Paw inflammation was be quantified based on paw swelling . The volume of the left hind  paw was be measured before and after arthritis induction by a mercury displacement method using a plethysmometer for rats. Paw swelling was be expressed as % inhibition of using following formula:


Evaluation:
a).For primary lesions:The percent inhibition of paw volume of the injected left paw over vehical control was be measured at day 7.
b).For secondary lesions:The percent inhibition of paw volume of the non-injected right paw over  control was be measured at day 21.

4.      Erythrocyte Sedimentation Rate ( ESR) [Mythilypriyaa Rajendran,1997,61, 1861 – 1878]
Higher erythrocyte sedimentation rate is indicator of inflammatory disorder.

Method:  WESTERGREN’S METHOD

Requirements:
1)Westergren’s pipette and Westergren’s stand (2) Oxalate bulb, (3) Sodium Citrate  solution

Westergren’s pipette: It has a bore of 3mm and a length of 300 mm. It is marked with graduations from 0 to 200. It is open from both the ends.

Westergren’s Stand: It can accommodate six pipettes. There is base containing six   grooves each containing a rubber cushion to prevent leakage of blood from Westergren’s pipette. The stand allows the pipettes to remain in exactly vertical position.

Procedure:
1)A sample of blood (about 3 ml) was be obtained by puncturing retro-orbital plexus and  was be mixed with 3.8% sodium citrate solution in proportion of four parts of blood to one  part of citrate soon. The mixing of blood was be done by rotating the sample gently between the palms of hands.
2) The blood was be sucked slowly up to the mark zero in the Westergren’s tube.
3) The tube was be set upright in the Westergren’s stand, taking care that no blood escapes.  The tube was be fixed with the help of screw cap.
4)At the end of one hour and two hours, the upper level of red blood cell column was be     read. It was be indicator of mm of clear plasma or ESR.

5.      Serum Rheumatoid Factor (RF) [Jung  Hyun-Ju,2005, 359–367]

Principle Of The Method:
The RF-Turbilatex is a quantitative turbidimetric test for the measurement of RF in human serum or plasma. Latex particles coated with human gammaglobulin are agglutinated when mixed with samples containing RF. The agglutination causes an absorbance change, dependent upon the RF contents of sample that can be quantified by comparison from a calibrator of known RF concentration.

Preparations:

Diluent (R1)

Tris buffer 20 mmol/L, pH 8.2

Sodium azide 0.95g/L

Latex (R2)

Latex particles coated with human gamma globulin, pH 7.4

Sodium azide 0.95g/L

RF-CAL

Calibrator. Human serum. The RF concentration is stated on the vial   label.

Optional

Ref.:  1102114 Control serum ASO/CRP/RF level L

Ref.: 1102115

Control serum ASO/CRP/RF Level H.

Reagents for RF test

For RF Calibrator:
RF Calibrator was be reconstituted with 2.0 ml of distilled water. It was be mixed gently and brought to room temperature for 10 minutes before use.

For Calibration curve (range from 20 to 160 IU/ml):
The following RF calibrator dilutions was be prepared in the normal saline. The concentration of the RF calibrator was be multiplied by the corresponding factor stated in table below to obtain the RF concentration of each dilution.

Calibrator dilution

1

2

3

4

5

6

Calibrator RF (ml)

  ---

100

10

90

25

75

50

50

75

25

100

---

Factor

0

0.1

0.25

0.5

0.75

1.0

For one point calibration (linear range up to 100 IU/ml):
Preparation for RF calibrator dilution: 30ml RF calibrator + 70 ml normal saline The RF calibrator concentration was be multiplied by 0.33 to obtain the RF concentration of the diluted calibrator.

Procedure:
1. The reagents and the photometer (cuvette holder) was be brought to 37°C.

2. The following assay conditions was be described:
Wavelength: 650 nm (600-650 nm)
Temperature: 37°C
Cuvette light path: 1 cm

3. The instrument was be adjusted to zero with distilled water.

4. The following reagents was be pipette into a cuvette and mixed.

5. Absorbance was be read after 2 minutes (A2) of the sample addition.


Ablank

A2 Calibrator/ Sample

NaCl 9.0 g/l (ml)

7.0

-

Calibrator or sample (ml)

-

7.0

R1: Diluent (ml)

0.9

0.9

R2: Latex (ml)

0.1

0.1

Calculations:

For calibration curve:
Calculated the absorbance difference (A2- Ablank reagent) of each point of the calibration curve and plotted the values obtained against the RF concentration of each calibrator dilution. Rheumatoid factor concentration in the sample was be calculated by interpolation of its (A2-Ablank reagent) in the calibration curve.

For one point calibration:
[(A2-Ablank reagent) sample] / [(A2-Ablank reagent) calibrator] X Diluted calibrator concentration = IU/mL RF

The reading of RF factor of all the groups obtained was be compared with the control animals and was be expressed as IU/mL RF.

Reference Values :
Normal values up to 20 IU/mL. Each laboratory should establish its own reference range.

6.      Serum C - reactive protein (CRP) [Mythilypriyaa Rajendran,1997,61, 1861 – 1878.]

Principle:
C-reactive protein is measured by turbidimetry method of analysis. Turbidimetry is the measurement of the reduction in light transmission caused by particle formation. Light transmitted in forward direction is detected. The amount of light absorbed by suspension of particles depends on specimen concentration and on the particle size. Solution requiring quantitation by turbidimetry is measured using visible spectrophotometers.

Reagents:

Reagent 1: Diluent
Tris Buffer 20 mmol/L
Sodium Azide 0.95g/L pH 8.2

Reagent 2: Latex reagent
Suspension of Latex particles coated with anti human CRP
Sodium Azide 0.95g/L

Reagent 3: CRP Calibrator
Lyophillized Human Serum
CRP concentration on label

Calibration:
The assay was be calibrated to the reference material CRM 470/RPPHS.

Reagent Preparation & Stability:

Working Reagent:  Contents of the Latex Reagent vial was be gently mixed and working reagent was be prepared as,
1 ml Latex Reagent + 9 ml Diluent
This working reagent was be stable for 30 days at 2-8° C.

CRP Calibrator: Contents of vial was be reconstituted with 1.0 ml of distilled water and kept for 10 minutes before use.
This CRP calibrator was be stable for 30 days at 2-8° C or 3 months at -30° C.

Samples:
Fresh serum is preferred, though samples stored for 8 days at 2-8° C or 3 months at -20° C may also be used.

Equipments required:
Photometer,
Liquid dispensing system
General laboratory equipments.

Procedure:

Assay protocol

Wavelength : 540 nm (530-550nm)
Blank : Distilled water
Temperature : 37° C
Cuvette Path Length : 1 cm

Dispense

Calibrator

Samples

Working Reagent

500 µL

500 µL

Calibrator

Sample

3 µL

3 µL

Reagents for CRP test
Blank reading was be adjusted with distilled water. Working reagent and Calibrator/Sample was be pipetted out as per the pipetting system. Absorbance was be measured immediately (A1) and exactly after 2 minutes (A2) of sample addition.

Auto analyzer was be suitably programmed in fixed time mode with a nominal delay of 10 sec and a read interval of 120 sec.

Calculation:
(A2-A1) sample       X Calibrator Concentration
(A2-A1) Calibrator

Reference Values
Normal values up to 6 mg/L.
Each laboratory should establish its own reference range.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

7.      Serum A/G ratio [Mythilypriyaa Rajendran,1997,61, 1861 – 1878.]

Estimation of Protein Contents:

Principle:
Proteins, in an alkaline medium, bind with the cupric ions present in the biuret reagent to form a blue-violet coloured complex; the intensity of the color formed is directly proportional to the amount of protein present in the sample.

Reagents:
Reagent A: Biuret reagent.
Reagent B: Protein standard.
Pipette into clean dry test tubes labeled as blank, standard and test.

Addition Sequence

Blank

Standard

Test

Biuret Reagent

1.0

1.0

1.0

Distilled water

0.02

-

-

Protein standard

---

0.02

-

Sample

---

---

0.02

Mix well and incubate at 37oC for 10 min. or at room temperature for 30 min. Measure the absorbance of the standard, test sample against the blank within 60 min at 550nm.

Calculation:
Total proteins in gm/ dl = (Abs T/ Abs S) X 8
Normal Range: 6-8gm/dl

Determination of Serum Albumin: (Godkar. 2005)

Name of the method: Bromocresol green method.

Principle: Albumin present in serum binds specifically with bromocresol green at pH 4.1 to form green colored complex, intensity of which can be measured colorimetrically by using 640 nm.

Normal Range: 3.3-4.8 g/dL.

Preparation of reagents:

1) Albumin reagent (ready to use): It is prepared by mixing following chemicals in 900    ml distilled water.
a) Succinic acid: 8.85 gm
b) Bromocresol green: 108 mg
c) Sodium azide: 100 mg
d) Brij-35: 4.0 ml

pH of this solution is adjusted by using 1 N sodium hydroxide to 4.1. Final volume is    made to one litre by using distilled water. It is stable at room temperature for one year.

2) Albumin standard (4gm/dl): Bovine albumin 4.0 gm in 100 ml of normal saline    containing 0.1 gm/dl sodium azide. It is stable at 2-8°C for one year.

Procedure:

Mono step method

  • Take three test tubes and labeled them as test, standard and blank.
  • Add 5 ml of albumin reagent in each of the test tube.
  • Then add 0.05 ml of serum, albumin standard and distilled water respectively.
  • Mix thoroughly and keep at room temperature (25°C+ 5° C) for exactly 10 minutes.
  • Measure the intensities of the test and standard by setting at 100% T, by using 640 nm (red filter).

Calculation:
Serum albumin g/dl = O.D. test/ O.D. Std. x 4

Determination of Globulins:
Normal range = 1.8-3.6 g/dl

A/G ratio = serum albumin (g/dl) / serum globulins (g/dl)
Normal range = 1.2:1 to 2:1

8.       Histopathological analysis [chemico biologicalinteraction, [ijpd, curative effect on rat]
The proximal interphalangeal joint of the hind paw of the rats were removed and separated from the surrounding tissues and weighed. The joints fixed in 10% formalin were decalcified, sectioned and finally stained with haematoxylin and eosin to examine the histopathological changes during the experimental period in all the groups under light microscope.

Cartilage and bone destruction by pannus formation was scored ranging from 0, no change; 1, mild change (pannus invasion within cartilage);  2, moderate change (pannus invasion into cartilage/subchondral bone); 3, severe change (pannus invasion into the subchondral bone); and vascularity (0, almost no blood vessels; 1, a few blood vessels; 2, some blood vessels; 3, many blood vessels). Histopathological  changes in the knee joints were scored in the femur region on 5 semiserial sections of the joint, spaced 70  μm apart. Scoring was performed on decoded slides by two observers, as described earlie

9.       Radiological analysis[chemico biologicalinteraction, [ijpd, curative effect on ra]
Before sacrificing the animals, X-rays were taken at the joints of the hind paw of the animals for evaluating the bone damage. Radiographs were taken using X-ray apparatus and industrial X-ray ?lm. The X-ray apparatuswas operated at 220Vwith a 40V peak, 0.2 s exposure time, and a 60 cm tube-to-?lmdistance for anterior–posterior projection.

The following radiograph criteria were considered: These scores (destroyed or intact joint) were used as a quantal test for bone necrosis. Radiographs were carefully examined and abnormalities were graded as follows:
(i)  Periosteaic reaction, 0 - 3 (none, slight, moderate, marked);
(ii)  Erosions, 0 - 3 (none, few, many small, many large);
(iii) Joint space narrowing, 0  - 3 (none, minimal, moderate, marked);
(iv) Joint space destruction, 0 - 3 (none, minimal, extensive, ankylosis).
Bone destruction was scored on the patella as described previously.

RESULTS

A.    Body Weight Change
The body weight of the control rats progressively increased while the gain in body weight of arthritic rat was retarded significantly. Drug treated group showed progressive increase in body weight.


Figure: Effect of methanolic extract of Capparis aphylla Roth. on % body   weight change.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups

B.     Arthritic Index
In this study, the incidence of inflammation was 100% on administration of freund’s adjuvant (i.e all the rats responded with an arthritic scored >1), if the rats were not treated with anti inflammatory agent. There was a difference in the severity of  inflammation being less in treated group and this could be attributed to the action of standard or MECA.


Figure :Effect of methanolic extract of Capparis aphylla Roth. on arthritic index.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups 

C.    Paw Volume
An intraplantar  injection of  CFA into the left hind paw produced a more marked inflammatory response in the model control animals than the normal control(Fig 4.4.3).Seven days after CFA injection, the edema volume were significantly higher in model control animal than normal control animals


Figure : Effect of methanolic extract of Capparis aphylla Roth. on paw edema.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups

D.    Erytrocyte Sedimentation Rate (ESR)
The indomethacin, MECA 190 mg/kg, MECA 240 mg/kg and MECA 300 mg/kg  treated groups ESR was significantly lower as compared to model control  group (Fig4.4.4).


Figure :Effect of methanolic extract of Capparis aphylla Roth. on ESR.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE

E.     Serum Rheumatoid Factor (RF)
A significant reduction in RF was observed in indomethacin, MECA 190 mg/kg, MECA 240 mg/kg) and MECA 300 mg/kg treated group as compared to model control group  (Fig 4.4.5).


Figure :Effect of methanolic extract of Capparis aphylla Roth. on serum rheumatoid factor.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups

F. Serum C-Reactive Protein (CRP)
A significant reduction in CRP was observed in indomethacin, MECA 190 mg/kg, MECA 240 mg/kg  and MECA 300 mg/kg treated group as compared to model control group.  (Fig4.4.6)


Figure : Effect of methanolic extract of Capparis aphylla Roth.on serum CRP.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups

G.    Serum albumin globulin ratio (A/G ratio) :
There was a significant decrease  in total protein albumin level, A/G ratio but significant increase in the level of the globulin by indomethacin in CFA inflammatory rats. the above change were brought back to near normal levels by MECA 190 mg/kg, MECA 240 mg/kg, MECA 300 mg/kg. Fig (4.4.7).


Figure :Effect of methanolic extract of Capparis aphylla Roth. on serum A/G ratio.

Values are expressed as mean ± SEM. N=6.Values are statistically evaluated using ANOVA analysis followed by Dunnett’s test. Significant values were compared with #P<0.05 normal control Vs model control & *P<0.05 model control Vs all MECA groups

H.    Effect of MECA on radiological changes of control and experimental animals  [IJPD, CURATIVE EFFECT ON RA]
Bone destruction, which is a common feature of adjuvant  arthritis, was examined by radiological analysis. Freund’s Complete Adjuvant treated rats had developed definite joint space narrowing of the intertarsal joints, diffuse soft tissue swelling that included the digits, diffuse demineralization of bone, marked periosteal thickening, and cystic enlargement of bone and extensive erosions produced narrowing or pseudowidening of all joint spaces and bending of phalangeal joints. In contrast, in rats treated with MECA  attenuate abnormalities consisted of asymmetric soft tissue swelling and small erosions, periosteal thickening, and minimal joint space narrowing, predominantly localized to the proximal areas of the paws. The drug control rats showed the normal architecture of phalangeal joints as that of control rats.

I.       Effect of MECA on histopathological changes of control and experimental animals[IJPD, CURATIVE EFFECT ON RA]
As shown in Fig  5 a  [NC]:Histology of synovial joint of normal control rat with intact morphology of synovium and synovial lining Fig  5 b [MC]: FA induced disease control rat showed plenty of lymphocytic infiltration [↓]in synovial lining with severe inflammation and marked angiogenesis [?] studied with proliferation of synovial cells [↓↓]Fig 5 c [SD]]: indomethacin 100mg/kg treated rats showed significant protection with mild lymphocytic infiltration [↓] with no evidence of thickening of synovial lining and angiogenesis Fig 5 d MECA 190mg/kg: MECA 190mg/kg treated rats showed milder angiogenesis [?], lymphocytic infiltration [↓] and synovial lining thickening[↓↓]. Fig 5 e MECA 240 mg/kg: MECA 240 mg/kg treated rats showed milder angiogenesis [?], synovial lining thickening [↓↓] with no evidence of lymphocytic infiltration. Fig 5 f MECA 300mg/kg: MECA 300mg/kg treated rats sho