In-Vitro Comparative Evaluation of Anti-Inflammatory Activity of Tinospora cordifolia and Ocimum sanctum

Abstract

The emergence of multidrug-resistant pathogens has increased the need for safer and effective alternatives to conventional anti-inflammatory drugs. Medicinal plants rich in phytochemicals offer promising therapeutic potential. The present study evaluates the in-vitro anti-inflammatory activity of aqueous extracts of Tinospora cordifolia (Guduchi) and Ocimum sanctum(Tulsi) using the protein denaturation method. Both plants have been traditionally used in Ayurveda for managing inflammation, fever, and various chronic disorders. The results showed significant inhibition of protein denaturation by the extracts, with maximum inhibition observed at higher concentrations. The combination of T. cordifolia and O. sanctum demonstrated greater activity (45.87% inhibition at 200 µg/ml) compared to individual extracts. These findings indicate that the synergistic use of these plant extracts could serve as a cost-effective and natural alternative for the management of inflammatory conditions.

Keywords

Introduction

Inflammation is a complex biological response of tissues to harmful stimuli such as pathogens, injury, or chemical irritants. While essential for host defense, prolonged or uncontrolled inflammation contributes to chronic diseases including arthritis, diabetes, cardiovascular disorders, and cancer [1].

The rising incidence of drug-resistant pathogens and side effects of synthetic anti-inflammatory drugs highlight the urgent need for novel, safer alternatives [2,3]. Medicinal plants, long used in Ayurveda and other traditional systems, provide an effective reservoir of bioactive compounds such as alkaloids, flavonoids, terpenoids, and phenolics with proven anti-inflammatory properties [4,5].

Tinospora cordifolia  (Guduchi) and Ocimum sanctum (Tulsi) are well-documented medicinal plants in Ayurveda. T. cordifolia is known for its immunomodulatory, antioxidant, and anti-inflammatory activities, while O. sanctum is revered as an adaptogen with antimicrobial, anti-stress, and anti-inflammatory effects. This study investigates the comparative in-vitro anti-inflammatory potential of their aqueous extracts.

AIM AND OBJECTIVES

Aim:

To evaluate and compare the in-vitro anti-inflammatory activity of aqueous extracts of Tinospora cordifolia  and Ocimum sanctum.

Objectives:

1. To prepare aqueous extracts of T. cordifolia and O. sanctum.
2. To evaluate anti-inflammatory activity by the protein denaturation method.
3. To compare the activity of individual extracts and their combination against the standard drug diclofenac sodium.

MATERIALS AND METHODS

Collection and Authentication

Leaves of Tinospora cordifolia  and Ocimum sanctum were procured from local markets in Andhra Pradesh, India, and authenticated by a botanist.

Preparation of Extracts

60 g of powdered leaves of each plant were subjected to aqueous extraction using 500 ml distilled water for 48 hours. Extracts were filtered using Whatman No.1 filter paper, concentrated, and stored in a desiccator for further use.

In-vitro Anti-inflammatory Activity

The activity was assessed using the protein denaturation method [10,11]:

• Reaction mixture: 0.2 ml egg albumin + 2.8 ml phosphate buffer saline (pH 6.4) + 2 ml plant extract at varying concentrations.
• Control: Distilled water.
• Incubation: 37 °C for 15 min, followed by heating at 70 °C for 5 min.
• Absorbance was measured at 660 nm.
• Diclofenac sodium was used as standard.

% Inhibition was calculated as:

% Inhibition = (Acontrol?−Asample?)?×100

             Acontrol?

Statistical Analysis

All experiments were performed in triplicate (n=3). Data are presented as mean ± standard deviation (SD). Statistical analysis was carried out using one-way analysis of variance (ANOVA), followed by Tukey’s post-hoc test to compare differences between groups. A p-value < 0.05 was considered statistically significant.

RESULTS AND DISCUSSION

Effect of T. cordifolia Extract

At 200 µg/ml, inhibition reached 41.35 ± 1.2%, which was statistically lower (p < 0.05) than diclofenac (46.3 ± 0.9%).

Effect of O. sanctum Extract

At 200 µg/ml, inhibition was 44.43 ± 1.0%, showing no significant difference (p > 0.05) compared with diclofenac (46.3 ± 0.9%).

Effect of Combination

The combined extracts exhibited maximum inhibition of 45.87 ±0.8% at 200 µg/ml. The effect was statistically higher than T. cordifolia alone (p < 0.05), but comparable with O. sanctum and diclofenac (p > 0.05).

These findings suggest a synergistic effect when both plant extracts are combined. Protein denaturation is a well-established mechanism in inflammation and autoimmune disorders [10,11]. The ability of extracts to inhibit denaturation highlights their anti-inflammatory efficacy. The results are consistent with previously reported pharmacological activities of T. cordifolia and O. sanctum[6–9].

CONCLUSION

The present study demonstrates that aqueous extracts of Tinospora cordifolia and Ocimum sanctum exhibit significant in-vitro anti-inflammatory activity. The combination of both extracts showed greater inhibition than individual extracts, indicating synergistic potential. Statistical analysis confirmed that the activity of the combination was significantly higher than T. cordifolia alone and comparable to diclofenac sodium. These results support the traditional use of these plants and suggest their application in developing safe, effective, and affordable herbal formulations for managing inflammation.

REFERENCES

1. Al-Bari MA, Sayeed MA, Rahman MS, Mossadik MA. Characterization and antimicrobial activities of a phenolic acid derivative produced by Streptomyces bangladeshiensis, a novel species collected in Bangladesh. Res J Med Med Sci. 2006;1:77–81.
2. Zy EA, Area A, Aam K. Antimicrobial activity of some medicinal plant extracts in Palestine. Pak J Med Sci. 2005;21:187–93.
3. Adriana B, Almodóvarl ANM, Pereiral CT, Mariângela TA. Antimicrobial efficacy of Curcuma zedoaria extract as assessed by linear regression compared with commercial mouthrinses. Braz J Microbiol. 2007;38:440–5.
4. Vijaya K, Ananthan S. Microbiological screening of Indian medicinal plants with special reference to enteropathogens. J Altern Complement Med. 1997;3:13–20.
5. Silver LL. Discovery and development of new antibiotics: the problem of antibiotic resistance. Antimicrob Agents Chemother. 1993;37:377–83.
6. Ferrero-Miliani L, Nielsen OH, Andersen PS, Girardin SE. Chronic interleukin-1β-driven inflammation: importance of NOD2 and NALP3 in its generation. Clin Exp Immunol. 2007;147(2):227–35.
7. Hannoodee S, Nasuruddin DN. Acute inflammatory response. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2020 [cited 2025 Sep 22]. PMID: 32310543.
8. Vogel WH, Berke A. Brief history of vision and ocular medicine. Amsterdam: Kugler Publications; 2009. p. 97. ISBN: 978-90-6299-220-1.
9. Porth C. Essentials of pathophysiology: concepts of altered health states. 2nd ed. Hagerstown (MD): Lippincott Williams & Wilkins; 2007. p. 270. ISBN: 978-0-7817-7087-3.
10. Brown JH, Mackey HK. Inhibition of heat-induced denaturation of serum proteins by mixtures of nonsteroidal anti-inflammatory agents and amino acids. Proc Soc Exp Biol Med. 1968;128(1):225–8.
11. Grant NH, Alburn HE, Kryzanauskas C. Stabilization of serum albumin by anti-inflammatory drugs. Biochem Pharmacol. 1970;19(3):715–22.