Research Articles

2020  |  Vol: 6(1)  |  Issue: 1 (January-February)  |  https://doi.org/10.31024/ajpp.2020.6.1.9
Comparative evaluation of anti-arthritic activity of Pongamia pinnata (L.) Pierre seeds and Bryophyllum pinnata (L.) leaves: In-vitro study

Divya Singh1*, Jai Singh Vaghela1, Pushpendra K. Saini2, Narendra Sharma2

1Bhupal Nobles College of Pharmacy, Udaipur, Rajasthan, India

2Sri Balaji College of Pharmacy, Benad Road, Jaipur, Rajasthan, India

*Address for Corresponding Author

Divya Singh

Bhupal Nobles College of Pharmacy, Udaipur, Rajasthan, India

 

Abstract

Objective: Comparative Anti-arthritic Activity of seed extract of Pongamia pinnata (L.) Pierre and Bryophyllum pinnata leaves by in vitro model. Materials and methods: Pongamia pinnata (L.) Pierre (Family: Leguminoseae) and Bryophyllum pinnata (Family: Crassulaceae) are a medicinal plant which is indicated for the treatment of arthritis in folklore medicine. The anti-arthritic activity of Pongamia pinnata seed and Bryophyllum pinnata leaves ethanolic extract were done by Inhibition of protein denaturation in vitro methods. The ethanolic extract of Pongamia pinnata seeds and Bryophyllum pinnata leaves were subjected to in vitro Inhibition of protein denaturation in various concentrations i.e. 50, 100, 200, 400, 800, 1000 and 2000μg/ml. Results: P. pinnata and B. pinnata both extracts exhibited a concentration dependent inhibition of protein (albumin) denaturation. The prepared extracts showed better anti-Arthritic activity than the standard drug; when compared with each other, P. pinnata extracts showed better anti- arthritic activity compared to B. pinnata extracts. Conclusion: In the present studyPongamia pinnata seed extract showed better activity comparative to Bryophyllum pinnata leaves and standard drug.

KeywordsPongamia pinnataBryophyllum pinnta, anti-arthritic activity


Introduction

Remedies are made from single or multiple herbs and minerals for various medical conditions like asthma, flu, diabetes, arthritis, heart disease, digestive problems, mental health and skin problems. Herbal medicines yielding about 25% of currently used crude drugs with another 25% derived from chemically altered natural products.

In recent years, there is increased research on traditional Ayurvedic herbal medicines on the basis of their known effectiveness in the treatment of ailments for which they have been traditionally applied (Agrawal and Paridhavi, 2007; Chitme and Patel, 2009; Gautam et al., 2013; Rajput et al., 2011).

Rheumatoid arthritis is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells and angiogenesis in affected joints, which ultimately leads to the destruction of cartilage and bone (Gautam et al., 2013). The common signs & symptoms often affects the wrist joints and the finger joints closest to the hand, also other parts of the body besides the joints and causes pain, swelling, stiffness, and loss of function in the joints (Mohan, 2005; Patwardhan et al., 2010).

Pongamia pinnata seeds have several chemical constituents such as Karangin, pongamol, pongagalabrone, and pongapin, pinnatin and kanjone. Pongamia pinnata seeds used in hypertension, skin ailments and rheumatic arthritis. Seed powder valued as a febrifuge, tonic and in bronchitis and whooping cough (Nadkarni, 1998; Yadav et al., 2011). Bryophyllum pinnatum leaf contains P-coumaric acid, Ferulic acid, Syringic acid, Bufadienolides- Bryophyllin A (bryotoxin) chemical constituents. Bryophyllum pinnatum use in  Neuropharmacological, CNS depressant effects, Anti-inflammatory, analgesic, antinociceptive and wound healing effect (Bhattacharjee, 2004).

Materials and methods

Collection and authentication of plant

The seeds of Pongamia pinnata and Bryophyllum pinnatum leaves were collected from local vendors of Jaipur. Identified and authenticated of the plants in the Department of Botany, University of Rajasthan, Jaipur.

Preparation of Ethanolic extract of seeds of P. pinnata and B. pinnatum leaves

Dried Seeds and leaves were powdered mechanically through mesh sieve. The powdered were first defatted with petroleum ether (40–60°C) and extracted with ethanol by continuous hot percolation method using Soxhlet apparatus. The filtrate of the extract was concentrated to dryness.

Inhibition of protein denaturation method

The following procedure was followed for evaluating the percentage of inhibition of protein denaturation:

Standard solution was prepared by addition of 2 ml of Egg albumin, 28 ml of phosphate buffer and various concentrations of standard drug (Diclofenac sodium) conc. of 50, 100, 200, 400, 800, 1000 and 2000μg/ml.

Control solution was prepared by addition of 2 ml of Egg albumin (from fresh hen’s egg), 28 ml of phosphate buffer (pH 6.4) and 20 ml distilled water.

Test solution containing 2 ml of Egg albumin, 28 ml of phosphate buffer and various concentrations of plant extracts (PPEE and BPEE) conc. of 50, 100, 200, 400, 800, 1000 and 2000μg/ml.

All of the above solutions were adjusted to pH 6.4 using a small amount of 1N HCl. The samples were incubated at 37°C for 15 minutes and heated at 70°C for 5 minutes. After cooling the absorbance of the above solutions was measured using UV-Visible spectrophotometer at 660nm their viscosity was determined by using Ostwald viscometer (Chandra et al., 2012).

The percentage inhibition of protein denaturation was calculated using the following formula (Sangeetha et al., 2011):

Where, Vt = absorbance of test sample, Vc = absorbance of control

Statistical analysis

Three replicates of each sample were used for each test to statistical analysis and the data were represented as Mean ± Standard Error Mean.

Results and discussion

The anti-arthritic effect of Pongamia pinnata ethanolic extract (PPEE), Bryophyllum pinnatum ethanolic extract (BPEE) leaf and diclofenac sodium were evaluated against the denaturation of egg albumin in vitro.

The result is summarized in (Table 1 and Figure 1). PPEE, BPEE and Diclofenac sodium were exhibited concentration dependent inhibition of protein (albumin) throughout the concentration range of 50 to 2000 μg/ml.

Table 1. In-vitro activity of PPEE, BPEE and diclofenac sodium (std drug) by inhibition of protein denaturation method

Conc. (μg/ml)

PPEE (%Inhibition) (±SEM)

BPEE (% Inhibition) (± SEM)

Diclofenac sodium (%Inhibition) (± SEM)

50

30±2.88

24.52±0.011

107.66±0.005

100

53.67±1.85

52.83±0.006

112.99±0.010

200

98.67±0.88

98.08±0.012

116.98±0.004

400

154.33±2.33

147.07±0.002

143.07±0.002

800

366±1.154

363.25±0.030

238.99±0.003

1000

601.67±1.20

580.50±0.005

382.38±0.003

2000

727.67±1.45

710.37±0.013

472.63±0.012

Figure 1. % inhibition v/s concentration of protein denaturation of PPEE, BPEE and Standard drug

 

 

The present findings exhibited a concentration dependent inhibition of protein (albumin) denaturation by Pongamia pinnata ethanolic extract (PPEE) seed and Bryophyllum pinnatum ethanolic extract (BPEE) leaf throughout the concentration range of 50 to 2000μg/ml. Diclofenac sodium (at the concentration range of 50 to 2000μg/ml) was used as reference drug which also exhibited concentration dependent inhibition of protein denaturation; however, the effect of diclofenac sodium was found to be less as compared with PPEE and BPEE. Inflammatory and arthritic disease well documented was caused by denaturation of tissue proteins.

Protein denaturation is a process in which protein lose their tertiary and secondary structure by application of external stress or compound such as strong acid or base, an organic solvent or heat most biological protein lose their biological function when denaturated (Megha et al., 2013; Gambhire et al., 2009; Marliyah and Ananthi, 2015). Production of auto-antigens in certain arthritic diseases may be due to denaturation of proteins in vivo. The mechanism of denaturation probably involves alteration I electrostatic hydrogen, hydrophobic and disulphide bonding. The increments in absorbance of plant extracts and standard drug to indicated the stabilization of albumin protein (Chandra et al., 2012; Sangeetha et al., 2011).

Conclusion

The present study was comparative evaluation of anti-arthritic activity of ethanolic Extract of Pongamia Pinnata (L.) Pierre seed and Bryophyllum pinnata leaves with standard drug. The higher percentage inhibition of albumin denaturation at the highest concentration of ethanolic extract of Pongamia Pinnata (L.) was 2000μg/ml.

Acknowledgements

The author thankful to guide Dr Jai Singh Vaghlea, for assistance and Bhupal Nobles College of Pharmacy, Udaipur for supporting this project.

Conflicts of interest: Not declared.

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