{"id":53441,"date":"2023-12-31T11:04:02","date_gmt":"2023-12-31T11:04:02","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=53441"},"modified":"2024-01-05T06:31:25","modified_gmt":"2024-01-05T06:31:25","slug":"in-vitro-anti-inflammatory-and-antioxidant-activity-of-seed-ethanolic-extract-of-pongamia-pinnata","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no4\/in-vitro-anti-inflammatory-and-antioxidant-activity-of-seed-ethanolic-extract-of-pongamia-pinnata\/","title":{"rendered":"In vitro Anti-Inflammatory and Antioxidant Activity of Seed Ethanolic Extract of Pongamia pinnata"},"content":{"rendered":"\n

Introduction\n<\/strong><\/p>\n\n\n\n

The functional food and nutraceutical potential of phytochemical therapeutics is generating a great deal of interest today 1<\/sup>. Bioactive compounds in these natural compounds contain anticarcinogenic action and provide several health-promoting benefits 2<\/sup>. Numerous medicinal plants are known to generate bioactives with antioxidant and antibacterial properties. The phenolic acids, which hinder the growth of infections and cause minimal damage to host cells, are also intriguing prospects for the development of novel antimicrobial medications. As a result, there is a rising interest in the development of several plant-derived medicines with different biological activities for the treatment of diverse infectious illnesses 3<\/sup>.<\/p>\n\n\n\n

Pongamia pinnata (L.) Pierre, a member of the Fabaceae family, is extensively dispersed Historically, the parts of P. pinnata were utilised in the indigenous medicinal systems of several cultures. It has been observed that several primary phenolic compounds are found in diverse plant sections 4,5<\/sup>. Karanjin, a furanoflavone extracted from this plant’s seeds, has superior therapeutic properties. Extracts of the seed oil of P. pinnata were active against both Gram-positive and Gram-negative bacteria 6,7<\/sup>.<\/p>\n\n\n\n

Oral mucosal lesions, also known as OML, are any abnormal changes that occur on the surface of the oral mucosa. These changes can manifest as pigmented, ulcerative, red and white characteristics, or any swelling or developmental fault variations. Oral mucosal ulceration is an inflammatory lesion caused due to disintegration of the oral epithelium. 8<\/sup>,9<\/sup>,6<\/sup> It has been established that medicinal plants that are abundant in a number of different chemical ingredients are quite beneficial in the treatment of OML. Several oral mucosal lesions are treated by anti-inflammatory and antioxidants like retinol, lycopene etc. 10<\/sup>. Upon inflammation, toxins are released that cause cell damage and the body produces substances that activate the immune system. About half of analgesics are anti-inflammatory medicines, which treat pain by reducing inflammation as opposed to opioids 11<\/sup>. Diclofenac is the typical anti-inflammatory medication, and its negative effects have been recorded 12<\/sup>. <\/p>\n\n\n\n

In turn, the antioxidant capacity is dependent on the total amount of polyphenolic compounds, essential oils, and other components 13<\/sup>. Herbs are regarded as an excellent source of natural antioxidants, although very little research has been conducted on their potential use as antioxidants 14,15<\/sup>. The majority of studies have been conducted on these leaves, with the exception of their anti-inflammatory properties. Therefore, the purpose of this study is to extract the essence of P. pinnata’s therapeutic properties using ethanol as a solvent. This study’s objective is to investigate the antioxidant and anti-inflammatory properties of the seed extracts of P.pinnata in invitro methods.<\/p>\n\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

Collecting and extracting plants <\/strong><\/p>\n\n\n\n

P. pinnata seeds were gathered in\nChennai, Tamil Nadu, India. For the extraction, shade-dried P. pinnata plant\nseeds were used. The ethanol extract was made by soaking the seed powder in\nwater and then evaporating it. The collection of dry powder for subsequent\nexamination. <\/p>\n\n\n\n

Evaluation of the phytochemical properties of P.pinnata seed extract <\/strong><\/p>\n\n\n\n

In this study, the content of phenols, tannins, saponins, proteins, and acids in the seed ethanolic extract of P.pinnata was examined qualitatively using phytochemical techniques (Harbone & Baxter, 1993). 16<\/sup><\/p>\n\n\n\n

Test for phenols <\/strong><\/p>\n\n\n\n

In a test tube, a tiny amount of\nethanolic extract is combined with 1 mL of water, 1 to 2 drops of iron III\nchloride (FeCl3) are added, and a black colour change is noticed. <\/p>\n\n\n\n

Test for tannins <\/strong><\/p>\n\n\n\n

3 drops of lead sub-acetate solution\nwere mixed with 1 ml of filtrate. creamy-coloured gelatin consistency implies\ntannins.<\/p>\n\n\n\n

Test for saponins <\/strong><\/p>\n\n\n\n

Foam test: After forcefully shaking\na 1 ml sample of the extract with water, persistent foam was detected. <\/p>\n\n\n\n

Test for proteins <\/strong><\/p>\n\n\n\n

One millilitre of ninhydrin was\ndissolved in one millilitre of acetone, followed by the addition of a little\namount of extract. The presence of proteins led to the development of a purple\nhue. <\/p>\n\n\n\n

Test for acids<\/strong><\/p>\n\n\n\n

This is measured by boiling 3.0 g of\nthe powder in 50 cc of 10% sulfuric acid for 5 minutes; the resulting ammonia\nis then freed, distilled into 0.1 N acid, and titrated. <\/p>\n\n\n\n

DPPH radical scavenging efficacy of ethanolic P.pinnata seed extracts <\/strong><\/p>\n\n\n\n

Using the approach of Hatano et al., the DPPH radical scavenging was evaluated (1989). Briefly, 1.0 ml of DPPH solution was added to 1.0 ml of extract at doses ranging from 0.1 to 0.5 mg\/ml. After 50 minutes at room temperature, the mixture’s activity was measured at 517 nm. As a standard, identical amounts of ascorbic acid were utilised. Using the following formula, the capacity to scavenge the DPPH radical was computed and represented as a percentage (%). <\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

In an Albumin denaturation\ninhibition assay, ethanolic seed extracts of P.pinnata demonstrate an\nanti-inflammatory effect in vitro. <\/p>\n\n\n\n

The anti-inflammatory efficacy was investigated using inhibition of albumin denaturation analysis, as determined by Leela Prakash and Mohan Dass’s approach (2010). The pH of the reaction mixture, which contained the test extracts as well as a 1% aqueous solution of bovine albumin fraction, was adjusted by adding a small amount of 1N hydrochloric acid. After heating the sample extracts for 20 minutes at 51 degrees Celsius after having been incubated at 37 degrees Celsius for 20 minutes, the turbidity was measured at 660 nanometers. The experiment was carried out three times. The proportion of protein denaturation inhibition was computed as follows: <\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Statistical analysis using a one-way\nANOVA in SPSS software v25, with a significance threshold of (p<0.05)\nconsidered.<\/p>\n\n\n\n

Results<\/strong><\/p>\n\n\n\n

In the present study, results of the DPPH radical scavenging activity of P.pinnata<\/em> seed ethanolic extract showed a significant (p<0.05) dose-dependent potential activity by inhibiting DPPH radical formation (figure 3). Inhibitory activity showed 10.74, 20.83, 36.71, 53.4 and 74.9% at 100, 200, 300, 400 and 500\u00b5g respectively. Protein denaturation inhibition activity of P.pinnata seed ethanolic extract showed a significant (p<0.05) dose-dependent potential anti-inflammatory activity (figure 4) such as 33,38,42,53 and 68% respectively whose activity was near to that of the standard drug diclofenac activity (14.8, 26.4, 37.1, 64.4 and 92%) and this study clearly indicates that P.pinnata seed extract exhibits a potential anti-inflammatory activity. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1: P. pinnata seed extraction preparation.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 2: Phytochemical screening of P.pinnata seed extract.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 3: In vitro antioxidant potential of P.pinnata seed ethanolic extract by DPPH radical scavenging activity<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 4: In vitro anti inflammatory potential of P.pinnata seed ethanolic extract by protein denaturation inhibition activity<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Discussion<\/strong><\/p>\n\n\n\n

In developing nations, pharmaceutical programmes continue to play a vital role as basic care therapeutic solutions. While phytochemical agents are predominantly used as a treatment for minor diseases at home, several medical practitioners like Siddha, Ayurveda and Unani have gained attention and shown promising cures for chronic diseases 17,18<\/sup>. Herbalists often do not treat acute psychological or physical illnesses; rather, the objective of herbal therapy is to promote long-term health improvements 19<\/sup>.<\/p>\n\n\n\n

The results were consistent with the previous study, which characterised P.pinnata leaf methanol extracts (400 mg\/kg) as effective anti-inflammatory agents, comparable to the present study 20,21<\/sup>. The DPPH test is a standard technique for measuring the ability of antioxidant molecules to scavenge free radicals by neutralising the stable, coloured DPPH radical. The radicals generated by DPPH are neutralised by plant extracts containing antioxidants, offering an ambitious foundation for future in vivo investigations. According to studies, phytochemicals such as phenolics and flavonoids can donate hydrogen and neutralise DPPH radicals 22,2324<\/sup>.<\/p>\n\n\n\n

Compared to the various concentrations, anti-inflammatory activities were clear in our investigation. Using the albumin denaturation assay, the strongest antiinflammatory effect was discovered at 50\u00b5l. In an in vitro investigation, Srinivasan et al assessed the antiinflammatory activity of a 70% ethanolic extract of P.pinnata leaves in rats and found significant antiinflammatory activity without any ulcerogenic potential. 25<\/sup>. Singh et al aimed to assess anti-inflammatory potential of ethanolic seed extract of P.pinnata in rats and found the best anti-inflammatory activity against Bradykinin and prostaglandin E1 induced inflammation and minimal effects against histamine and serotonin induced inflammation 26<\/sup>. Rekha et al also studied the characterization and anti-inflammatory properties of P.pinnata seed extract and found a significant increase in antioxidant and lipoxygenase inhibitory activity of ketone and oxide. The derivatives of P.pinnata showed higher anti-inflammatory activity in rat models 7<\/sup><\/a>. Dinesh et al showed that a herbal combination of 3 herbal plants showed superior antioxidant and antiinflammatory activities 27,28<\/sup>. <\/p>\n\n\n\n

Several oral lesions are treated by topical application of gels and topical agents with  antiinflammatory and antioxidant properties. Phytochemical agents like curcumin, aloe vera, etc show superior antioxidant and antiinflammatory properties29<\/sup>. Curcumin is predominantly studied its therapeutic use of oral lesions and wound healing 30,31<\/sup>. Plants like P.pinnata which are commonly and easily found and are being less explored in the field of medical research. Hence, an attempt was made to study its antioxidant and antiinflammatory properties. The presence of metabolites like tannins, saponins, acids, phenols and proteins in P.pinnata is the reason for high anti-inflammatory and antioxidant properties. Further research is needed to implement the ethanolic seed extract of P.pinnata as a therapeutic agent for oral mucosal lesions.<\/p>\n\n\n\n

Conclusion<\/strong> <\/p>\n\n\n\n

It was obvious from the investigation that the ethanolic seed extract of P.pinnata contained both antioxidant and anti-inflammatory properties. The anti-oxidant and anti-inflammatory properties of P.pinnata seed extract were considerably higher at 500\u00b5g, as shown by the DPPH test and suppression of albumin denaturation, respectively. Additional research is required to confirm the therapeutic effects of these plant extracts. Further extensive research on the utilisation of P.pinnata seeds in the treatment of oral mucosal lesions using this plant’s bioactives in in-vivo models is required in order to develop particular applications and synthesise novel and powerful medications of natural origin. <\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

We thank BPG Lab, Saveetha Dental College and Hospitals, SIMATS for the use of their research laboratory facilities. <\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

No conflicts of interest with regard to the current work. <\/p>\n\n\n\n

Funding Sources <\/strong><\/p>\n\n\n\n

There is no funding Sources.<\/p>\n\n\n\n

References <\/strong><\/p>\n\n\n\n

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