INTRODUCTION:

Man depends upon plants for his entire essentials requirement like food, clothing and shelter. Additionally, plants are a significant source of fine chemicals, which are used in the global pharmaceutical industry. Since many years ago, plants have been the traditional source of both raw materials and finished medicines. The majority of medicines used to cure ailments come from plants. Many studies on these natural products have been conducted during the previous few decades.

Scientific research on several traditional treatments has produced real medications. Several modern formulations also contain natural ingredients. The most significant medications derived from plant sources and currently used in therapeutic settings are ashwagandha, rauwolfia cinchona, opium, ergot, etc. Before being introduced to modern medicine, they were all recognised as healers in traditional medicine. Researchers are increasingly interested in identifying the active ingredients in their extracts with thorough follow-up examination of their mechanisms of action because it is impossible to ignore the medical efficacy of Indian traditional medicine.

The word inflammation obtained from the Latin word inflamers which means state of being inflame or heat associated with redness and swelling. This is a complex, integrated host response found only in vertebrates¹. Inflammation is either acute or chronic. Acute inflammation may be an initial response of the body to harmful stimuli. In chronic inflammation, the inflammatory response is out of proportion resulting in damage to the body²

There are various medicines for controlling and suppressing inflammatory crisis; steroidal (SAIDs) and non-steroidal anti-inflammatory drugs (NSAIDs) and immunosuppressant are the examples of these medications which are associated with adverse effects while in practice our goal is to apply minimum effective dose by the highest efficacy with the least adverse effects. Thus, we need to apply natural anti-inflammatory factors within medication therapy to achieve increased pharmacological action and the lowest degree of unwanted side effects.^3-5

Herbal medicines are promoting subjects in medicine and, of course, we have to increase our knowledge about them. Search for safe and effective anti-inflammatory agents have been given priority in scientific research in herbal system of medicine. Tamilnadia uliginosa (Rubiaceae), also known as adavi manga or devathamalle in Telugu, devine jasmine in English, wagatta in Tamil, and bharani in Hindi, is a member of the Rubiaceae family. It is a native of Thailand, Vietnam, Sri Lanka, Bangladesh, and India.

Tamilnadia uliginosa is a plant that is used in traditional medicine to treat astringent, diuretic, and other disorders like dysentery and diarrhoea. To our knowledge, no research has been done on the ethanolic extract of Tamilnadia uliginosa fruits' effectiveness in reducing inflammation, though. So, the purpose of this study was to evaluate the anti-inflammatory activity.⁶

MATERIALS AND METHODS:

The methodology adopted to evaluate the anti-inflammatory activity of fruits of Tamilnadia uliginosa plant here under.

Collection and Authentication of plant material:

The fruits of Tamilnadia uliginosa were collected from the Talakona regions of chittoor dist, Andhra Pradesh and India. The plant material was taxonomically identified by botanist Dr. Madhava Chetty, Asst. Prof, SVU, Tirupathi. A voucher specimen has been preserved in our laboratory for future reference. The plant material was dried in shade, pulverized, passed through sieve no. 40 and stored in air tight container and used for further extraction.

Preparation of plant extract:

The fruits were cleaned and dried under standard shade conditions. Then it was grinded to obtain coarse powder of standard size suitable for extraction. The fruit powder was subjected to maceration. After that the extract was defatted with petroleum ether to remove the fats. Finally the extract was air dried. The practical percentage yield will be calculated.

Phytochemical screening^7-11

The extract was subjected to preliminary phytochemical screening in order to detect the presence of various groups of phytoconstituents by using standard phytochemical procedures.

Evaluation of in-vitro anti-inflammatory activity by

A). Inhibition of albumin denaturation method¹²

The anti inflammatory activity of Tamilnadia uliginosa was studied by using inhibition of albumin denaturation technique which was studied according to Mizushime et al and Sakat et al followed with minor modifications. The reaction mixture was consists of test extracts and 1% aquous solution of bovine albumin fraction, pH of the reaction mixture was adjusted using small amount of 1N Hcl. The sample extracts were incubated at 37°C for 20min and then heated to 51°C for 20min, after cooling the samples the turbidity was measured at 660nm. (UV-Ltd) The experiment was performed in triplicate.

The Percentage inhibition of protien denaturation was calculated as follow:

Percentage inhibition = (Abs Control-Abs Sample) X100/ Abs Control.

B). Antiproteinase action method:

The test was performed according to the modified method of Oyedepo et al and Sakat et al.The reaction mixture(2ml) was containing 0.06mg trypsin, 1ml 20Mm Tris HCl buffer (Ph7.4) and 1ml test sample of different concentrations (100-500 μg/ml).The mixture was incubated for an additional 20min.2ml of 70% perchloric acid was added to arrest the reaction. Cloudy suspension was centrifuged and the absorbance of the supernatant was read at 210nm against buffer as blank. The experiment was performed in triplicate.

The Percentage inhibition of proteinase inhibitory activity was calculated.

Percentage inhibition = (Abs Control-Abs Sample) X100/ Abs Control.

RESULTS AND DISCUSSION:

Table 1: Phytochemical screening of Tamilnadia uliginosa fruits

S. No	Chemical constituents	Ethanolic extract
1	Carbohydrates	Present
2	Proteins	Present
3	Amino acids	Absent
4	Tannins	Present
5	Glyosides	Absent
6	Alkaloids	Present
7	Flavonoids	Present
8	Steriods	Present
9	Saponins	Present

In vitro-Anti-inflammatory effect of Tamilnadia uliginosa fruits by Albumin denaturation method:

Table 2: Albumin denaturation % inhibition

S. No	Concentration (μg/ml)	Test absorbance	Standard absorbance	% Inhibition Test Std
1	100 μg/ml	0.085	0.075	54%	59.6%
2	200 μg/ml	0.068	0.053	63.4%	71.5%
3	3100 μg/ml	0.065	0.029	65.05%	84.4%
4	400 μg/ml	0.063	0.022	66.12%	88.1%
5	500 μg/ml	0.057	0.020	69.35%	89.2%

Figure:1 % inhibition graphical representation of In vitro- anti-inflammatory activity of Tamilnadia uliginosa fruits by Albumin denaturation method.

Protein denaturation is a process in which proteins lose their tertiary structure and secondary structure by application of external stress or compound, such as strong acid or base, a concentrated inorganic salt, an organic solvent or heat. Most biological proteins lose their biological function when denatured. Denaturation of proteins is a well documented cause of inflammation. As part of the investigation on the mechanism of the anti-inflammation activity, ability of plant extract to inhibit protein denaturation was studied. It was effective in inhibiting heat induced albumin denaturation. Maximum inhibition of test sample 69.35% was observed at 500μg/ml. Aspirin; a standard anti-inflammation drug showed the maximum inhibition 89.2% at the concentration of 500μg/ml compared with control. Each value represents the mean±SD. N=3, Experimental group were compared with control **p<0.01, considered extremely significant.

In vitro-Anti-inflammatory effect of Tamilnadia uliginosa fruits by Anti proteinase action:

Table 3: Anti proteinase % inhibition

S. No	Concentration (μg/ml)	Test absorbance	Standard absorbance	% Inhibition Test Std
1	100 μg/ml	0.042	0.034	53.3%	62.2%
2	200 μg/ml	0.035	0.026	61.1%	71.1%
3	300 μg/ml	0.032	0.018	64.4%	80%
4	400 μg/ml	0.030	0.012	66.6%	86.6%
5	500 μg/ml	0.026	0.009	71.1%	90%

Figure 2 % Inhibition graphical representation of In vitro- anti-inflammatory activity of Tamilnadia uliginosa fruits by Anti proteinase method.

Serine proteinase is known to be abundant in neutrophils, which are found at lysosomes. Leukocyte proteinase has been implicated in the development of tissue damage during inflammatory reactions, and proteinase inhibitors have been shown to provide a significant level of protection. Table 3 displays the antiproteinase activity of the extract at various concentrations. At 500 g/ml, it showed a maximum inhibition of 71.1% for the test sample. At 500g/ml, aspirin exhibited a 90% maximal inhibition. Each value represents the mean ± SD. N=3, Experimental group were compared with control **p<0.001, considered extremely significant *p<0.005.

CONCLUSION:

Plants are one of the most important sources of medicines. Since ancient times, medicinal plants have been used to treat a variety of diseases because to their availability, cost, inherited practise, economic feasibility, and perceived efficacy. The study's findings suggest that the ethanolic extract from Tamilnadia uliginosa fruit exhibits anti-inflammatory properties in vitro, which may be due to the presence of phyto-constituents like flavonoids, tannins and phenols, and saponins. It may be advisable to do additional in-vivo study and isolate the active compounds in order to fully investigate the possibilities.

CONFLICT OF INTEREST:

We declare that we have no conflict of interest.

ACKNOWLEDGMENT:

We wish to thank Management and Dr. M. Chinna Eswaraiah garu, Principal of Anurag Pharmacy College, Kodad for their moral support and blessings and also express our heartful thanks to Dr. Madhava Chetty for authentication of plant material.

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Received on 15.07.2020 Modified on 25.02.2022

Research J. Pharm. and Tech 2023; 16(4):2022-2024.

DOI: 10.52711/0974-360X.2023.00332