INTRODUCTION:

Inflammation generally takes place once causative agents infest the body, remain in a specific tissue and/or disperse in the blood. In most cases, the development of inflammation occurs by the involvement of the natural immune response along with the acquired immune response¹. Inflammation is a serious reaction by the body tissue to all types of injuries. Wound, hyperemia, warmth and lump may be the four key indicators of inflammation. Blood circulation will rise towards the injured area, because the blood vessels surrounding the tissue expand when there is a wound in any body part².

The number of factors like a disease-causing agent, oxygen starvation, necrobiosis, cancer, mutual action of antigen-antibody and thermal cycling might be at the origination of the inflammatory process.

Immunosuppressive agents, steroid and non-steroidal drugs are commonly used to relieve inflammatory disease with prolonged use in which the usage of these drugs results in adverse effects as an analogy of peptic

ulcer, cardiovascular disorder, impairment of renal function and bleeding gastrointestinal. This has given rise to look for alternative therapies. From this perspective, natural products obtained from herbal plants have been proven to help treat inflammation and pain^1,2,
3. A day-by-day research study is increasing in understanding the mechanisms involved in causing inflammation and also about the association of free radicals in the development of inflammation³.

Around 80 percent of the population living in underdeveloped countries chooses to make use of ancestral therapeutic practices for their health care system⁴. An attempt has been made by WHO (World Health Organization) in the identification of pharmacological important plants using globally in which above 20,000 species are listed. Majority of the plant parts are used as raw medicine as it holds several pharmacological qualities². The medicinal plant's requirements are rising owing to the increasingly being recognized as the producer of herbal products possessing lesser adverse effects, readily available and make relatively cost-effective⁵. An herbal product finds its application in various fields like the medical industry, agricultural industry, cosmetic and food industry. In the history of all civilizations, it has been well documented that medicinal plants are practice for treating diseases⁶. Investigation of phytochemicals in medicinal herbs is extremely essential in determining a novel source of medicinally and industrially significant metabolites. It is crucial to start important measures for the screening of secondary metabolites production from plants⁷.

The Cucurbitaceae family is an uncommon family of dicotyledons, generally recognized as the gourd family and it is considered to be a great source in the pharmacy industry with several cost-effective species. Due to this fact, researchers are focusing on this family^8,4. Cucurbitaceae consists of 125 genera and 960 species which are considered to be the greatest family among summer vegetable crops⁹. The biochemical compound cucurbitacins are present in this family which is an immensely bitter and harmful substance and it is considered to be a strong candidate in several pharmacopeias⁸. Since ancient times, it is a well-known fact that the habit of using plant or their extracts for treating inflammation¹. The present study was aimed to evaluate the anti-inflammatory activity of leaf extracts by protein denaturation assay.

MATERIALS AND METHODS:

Sample collection and extraction preparation:

Fresh and healthy leaves of Momardica charantia (MC), Citrullus lanatus (CL), Coccinia grandis (CG) and Lagenaria siceraria (LS) were collected from Vellore district, Tamil Nadu, India. The leaves were washed in double-distilled water and shade dried for 3-5 weeks. Then the leaves were grounded to a fine powder and stored in an airtight container for further use. About 10g of leaf powder was taken in a flask and dissolve with 200ml of methanol and petroleum ether solvents and kept in a shaker for 2 days at 120rpm. The content was filtered using Whatman No.1 filter paper and allowed to evaporate to obtain crude extract for further study¹⁰.

Protein denaturation assay:

The 5ml of the reaction mixture contains 1% bovine serum albumin (0.2ml), phosphate-buffered saline at pH6.4 (4.78mL) and the plant extract (0.02mL) and then the content was blended and incubated at 37^◦C for 15 minutes in a water bath. After incubation time it was heated for 5 minutes at 70^◦C. Then the reaction mixture was allowed to cool at room temperature and its turbidity was measured using a UV visible spectrometer at 660nm. PBS (Phosphate Buffer Saline) solution was used as the control. Aspirin was used as the standard¹¹.

The experiment was done in triplicates and the percentage of protein denaturation inhibition was calculated using the formula:

Percentage of inhibition= absorbance control- absorbance sample/ absorbance control ×100

RESULT AND DISCUSSION:

The present study concludes that both the extracts of Momardica charantia, Citrullus lanatus, Coccinia grandis and Lagenaria siceraria showed significant anti-inflammatory activity in a dose-dependent manner. As the concentration of the extracts increased the inhibition percentage also increased as shown in Figures 1 and 2. The inhibition range was shown between 10-77% for the concentrations of 100-300 µg/mL.Worldwide it is an important need in finding anti-inflammatory plants for drug formulation since inflammation is a prevalent health problem wherewith existing pharmacological therapies are very little¹². Petroleum ether crude extracts showed a higher inhibition percentage compare to methanol crude extracts. Coccinia grandis has shown maximum inhibition percentage compare to the other three petroleum ether extracts.

Figure 1: Protein inhibition assay of methanol leaf extracts (MC- Momardica charantia, CL- Citrullus lanatus, CG-Coccinia grandis and LS- Lagenaria siceraria)

Figure 2: Protein inhibition assay of petroleum ether leaf extracts (MC- Momardica charantia, CL- Citrullus lanatus, CG-Coccinia grandis and LS- Lagenaria siceraria)

A previous study reported that anti-inflammatory activity was showed by aqueous leaf and stem extract of Coccinia grandis against formaldehyde-induced rat paw edema¹³. Immune cells take part in inflammation-based reactions. Several studies showed that numerous extracts of Momardica charantia can serve as an immunomodulatory agent and, thereby, can also serve as an anti-inflammatory agent⁹. Coccinia grandis are useful in treating diabetes, high blood pressure, inflammation, tussis, diarrhea and blood purification³. This is the first report on comparison of anti-inflammatory activity of most commonly used plant belonging to this family which gives an idea in the pharmaceutical sector for the selection of plants for anti-inflammatory based drug design. The presence of phytoconstituents such as flavonoids, carotenoids and polyphenols in the leaf extracts could be responsible for this protein denaturation assay in which further in-vivo study should be done to validate these plants in the pharmacological industry.

CONCLUSION:

The results obtained in the present study allow us to conclude that the leaf extracts of Momardica charantia, Citrullus lanatus, Coccinia grandis and Lagenaria siceraria plants have potentiality in the development of new medication against inflammatory diseases and also it turns out to be an alternate bioresource for generating anti-inflammatory agents. World-wide this is an extremely significant discovery since inflammation is a prevalent medical issue in which the existing pharmacological therapies are insufficient. The leaves used in this study are uncommercialized, readily and sufficiently available for the whole year. However, detailed research needs to be done to represent the mechanism responsible for this activity and its biologically active metabolites.

ACKNOWLEDGEMENT:

The authors are thankful to Vellore Institute of Technology,Vellore for providing lab facilities for carrying out this study.

CONFLICT OF INTEREST:

Declared none

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Received on 21.02.2021 Modified on 30.12.2021

Research J. Pharm. and Tech 2023; 16(4):1598-1600.

DOI: 10.52711/0974-360X.2023.00261