Pharmacological Evaluation of Anti-inflammatory Activity of Nyctanthes arbor-tristis in Experimental Animal Models

Inflammation serves as the body's defense mechanism against harmful agents, including pathogens, damaged cells, or irritants. This response entails a series of intricate biochemical processes driven by inflammatory mediators like prostaglandins, leukotrienes, cytokines, and nitric oxide [1]. Acute inflammation is marked by the widening of blood vessels, heightened permeability of blood vessels, and the infiltration of white blood cells, while chronic inflammation is associated with tissue damage and the body's repair processes [10].

The activation of phospholipase A2 triggers the liberation of arachidonic acid, which is subsequently converted into prostaglandins and leukotrienes—essential mediators of inflammation. Nitric oxide (NO), generated by inducible nitric oxide synthase (iNOS), also significantly contributes to inflammatory processes.

Non-steroidal anti-inflammatory drugs (NSAIDs) like diclofenac function by blocking cyclooxygenase (COX) enzymes. Nonetheless, extended use of NSAIDs is linked to negative effects such as gastric ulcers, kidney damage, and cardiovascular issues. This has sparked a growing interest in plant-based alternatives that offer improved safety profiles.

Nyctanthes arbor-tristis, also known as Parijat, is a medicinal plant in the Oleaceae family. It is commonly employed in traditional medicine to treat inflammatory conditions like arthritis, fever, and joint pain [4,5]. Phytochemical analyses have identified the presence of iridoid glycosides, flavonoids, alkaloids, and tannins, which are recognized for their anti-inflammatory and antioxidant effects.

 Previous research indicates that the plant extract can inhibit inflammatory mediators such as TNF-α, IL-1β, and prostaglandins, as well as lower oxidative stress. Nonetheless, a thorough pharmacological assessment using standard experimental models is necessary to confirm its effectiveness. Therefore, the present study was designed to evaluate the anti-inflammatory activity of Nyctanthes arbor-tristis using dextran-induced and formalin-induced paw edema models in mice

Figure 1: Nyctanthes arbor-tristis

5. MATERIALS AND METHODS

5. 1. Experimental Animals

Healthy adult Manly mice (20 – 25 g) were used for the study. The creatures were housed under controlled environmental conditions (temperature 25 ± 1 °C, relative moisture 55 – 65, and 12- hour light/ dark cycle). Standard bullet diet and water were handed ad libitum.

All experimental procedures were carried out in agreement with guidelines of the CPCSEA and approved by the Institutional Animal Ethics Committee (IAEC blessing No. 751/ PO/ Re/ S/ 03/ CPCSEA/2026/1-1) (6).

5. 2. Plant Material and Authentication

The flowers of Nyctanthes dome- tristis were collected from original sources and authenticated by a good botanist from the institutional botany department. The factory material was gutted, shade- dried, and coarsely pulverized for farther processing.

5. 3. Preparation of Extract

The dried powdered factory material was subordinated to Soxhlet birth using 90 ethanol as detergent. The birth process was carried out for several cycles until complete birth was achieved. The attained excerpt was concentrated using a water bath and stored in a watertight vessel at room temperature for farther use.

 5. 4. Phytochemical Screening

Primary phytochemical webbing of the ethanolic excerpt was performed to descry the presence of colorful bioactive ingredients similar as flavonoids, alkaloids, tannins, phenolic composites, glycosides, and triterpenoids using standard qualitative tests. The presence of these ingredients was verified grounded on characteristic color changes and precipitate conformation.

5. 5. Acute Toxicity Study

Acute oral toxin study was conducted as per OECD guideline 425. The excerpt was set up to be safe at advanced boluses, and experimental boluses (200 mg/ kg and 400 mg/ kg) were named for pharmacological evaluation.

5. 6. Experimental Design

A total of 30 mice were divided into five groups (n = 6):

Table 1: Experimental design of animals

5. 7. Induction of Inflammation

5. 7. A. Dextran-Induced Paw Edema

Acute inflammation was convinced by edging in 0.1 mL of 1 dextran result into the sub-plantar region of the right hind paw. Paw volume was measured at different time intervals (0,1, 2, and 3 hours). The increase in paw volume indicated the extent of inflammation (7,8).

5. 7. B. Formalin- Induced Paw Edema

Inflammation was convinced by edging in 0.1 mL of 2 formalin result into the paw. This model produces biphasic seditious response involving neurogenic and seditious intercessors. Paw edema was measured up to 4 hourspost-induction (7,8).

 5. 8. Evaluation Parameters

 5. 8.1 Paw Edema Measurement

 Paw volume was measured using a plethysmometer. The degree of inflammation was

 calculated as the increase in paw volume compared to birth.

% inhibition of edema was calculated using the formula:

%Inhibition = [1− (????????−????????) /(????????−????????)] ×100

Where:

 Vt= treated group

 Vc = control group

 Vd = disease group 

5. 8.2 Locomotor exertion

Locomotor exertion was assessed using a Rota- Rod outfit at 25 rpm with a cut- off time of 180 seconds. Reduction in locomotor exertion indicates inflammation- convinced discomfort, while enhancement suggests remedial effect.

 5. 8.3 C- Reactive Protein (CRP)

Blood samples were collected from the tail tone, and serum was separated by centrifugation.CRP situations were measured using ELISA accoutrements Elevated CRP situations indicate systemic inflammation.

 5. 8.4 Body Weight

Body weight was recorded ahead and after treatment. Reduction in body weight indicates complaint progression, while enhancement reflects remedial effect.

 5. 8.5 Feed Input

Feed input was measured daily. Reduced input is associated with inflammation, while normalization indicates recovery.

5. 9. Statistical Analysis

All data were expressed as mean ± SEM (n = 6). Statistical analysis was performed using one-

 way ANOVA followed by Duncan’s multiple range test. Values were considered statistically

 significant at * p< 0.05 and p< 0.01

6. Results

6.1 Phytochemical Screening

Table 02: Phytochemical constituents of ethanolic extract of Nyctanthes arbor-tristis

6.2 Dextran-Induced Paw Edema

Table 03: Effect on paw edema (Dextran model)

Figure 02: Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis on dextran induced paw edema in mice.

6.3 Formalin-Induced Paw Edema

Table 04: Effect on paw edema (Formalin model)

Figure 03- Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on formalin-induced paw edema in mice.

6.4 Locomotor Activity

Table 05: Locomotor activity (Dextran model)

Fig 04. Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on Locomotor activity in Dextran-induced mice.

6.5 Locomotor Activity

Table 06: Locomotor activity (Formalin model)

Fig 05. Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on Locomotor activity in Formalin-induced mice.

6.6 C-Reactive Protein (CRP)

Table 07: CRP levels (Dextran model)

Figure 06: Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on C Reactive Proteins in Dextran-induced mice

6.7 C-Reactive Protein (CRP)

Table 08: CRP levels (Formaline model)

Fig 07. Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on C Reactive Proteins in Formalin-induced mice.

6.8 Body Weight

Table 09: Body weight (Dextran model)

Fig 08: Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on Body Weight in Dextran-induced mice.

6.9 Body Weight

Table 10: Body weight (Formaline model)

Fig09. Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on Body Weight in Formalin-induced mice.

6.10. Feed Intake

Table 12: Feed intake (Dextran model)

Fig 10. Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on Feed Intak in Dextran-induced mice.

6.11. Feed Intake

Table 13: Feed intake (Formaline model)

Fig 11. Effect of oral administration of ethanolic extract of Nyctanthes arbor-tristis (EENAT) on Feed Intake in Formalin-induced mice.