Research Article Evaluation of anti-inflammatory action of Illicium verum - An in vitro study Rachel Paul1, R. V. Geetha2* ABSTRACT Introduction: Illicium verum is a medium-sized evergreen tree native to northeast Vietnam and southwest China. A spice commonly called star anise. Star anise refers to aromatic herbs which are used in cooking for their distinctive flavor and their fragrance. Star anise is the major source of the chemical compound, shikimic acid which is a pharmaceutical synthesis of anti- influenza drug. It also has raw materials needed for fermentation of the food. Star anise has anti-inflammatory, antimicrobial, antifungal, and antioxidant properties. It has many medicinal properties which can also be used to treat cancer as well as gastric problems. It is an easily available herb in the market and is easily affordable by many people; it can be used in the treatment of various diseases. Materials and Methods: The anti-inflammatory activity was studied using protein denaturation assay and the results were read spectrophotometrically. Results: The anti-inflammatory activity of the extract was studied by its ability to inhibit protein denaturation. It was effective in inhibiting heat induced albumin denaturation at different concentrations. Maximum inhibition, 77.87 ± 1.55 was observed at 500 µg/ml. Half-maximal inhibitory concentration value was found to be 105.35 ± 1.99 µg/ml. Conclusion: The result obtained was compared to the commonly available nonsteroidal anti- inflammatory drugs such as aspirin. This research conducted to make the herb worthy for further investigation. KEY WORDS: Anti-influenza, Illicium vernum, Inflammatory, Nonsteroidal anti-inflammatory drug, Shikimic acid, Star anise INTRODUCTION Star anise is one of many spices that contain bioactive compounds as well as a number of phenolic Illicium verum belonging to the Magnoliaceae and flavonoid compounds, having antioxidant, family, commonly known as Chinese star anise is one preservative, and antimicrobial properties.[4] Star of the flavors used in China 5 spices, cultivated in anise has carminative, antispasmodic, antiseptic, mountainous region especially in Lanson province, antimicrobial, antidiarrheal activities and is used [1] Cochin, China (Southern China), and Vietnam. to treat colics and as a tranquilizer. Researchers I. verum, which originated in southwest China and attribute these effects to the presence of two coumarin northeast Vietnam, is an evergreen plant. I. verum derivatives: 7-hydroxycoumarin and 7-methoxy- fruits have traditionally been used as flavors. coumarin. However, star anise also contains a Simultaneously, I. verum was also used in medicine dicycloether, and anethole, both of which have [2] for a long time. The I. verum fruits are capsule- antimotility activities. However, the consumption of like aggregate with star-shaped five to 10 pointed high amounts of star anise tea may cause neurotoxicity boat shaped section about on eight averages. Each and hepatotoxicity.[5] arm is a seed pod. The fruits have tough skin and rust colored outer portion and seeds with high oil Traditional uses of I. verum are recorded throughout [3] content. Asia and Northern America, where it has been used for more than 10 types of disorders. Numerous Access this article online compounds including volatiles, seco-prezizaane- type sesquiterpenes, phenylpropanoids, lignans, Website: jprsolutions.info ISSN: 0975-7619 flavonoids, and other constituents have been 1Department of Microbiology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India, 2Department of Microbiology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India *Corresponding author: R. V. Geetha, Department of Microbiology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India. Phone: +91-9710456203. E-mail: [email protected] Received on: 14-05-2018; Revised on: 18-06-2018; Accepted on: 24-07-2018 Drug Invention Today | Vol 10 • Issue 12 • 2018 2441 Rachel Paul and R. V. Geetha identified from I. verum. Modern pharmacology saline (pH 6.3) was added to each tube. Turbidity studies demonstrated that its crude extracts and active was measured spectrophotometrically at 660 nm. compounds possess wide pharmacological actions, For control tests, 0.05 ml of distilled water was used especially in antimicrobial, antioxidant, insecticidal, instead of extracts while product control tests lacked analgesic, sedative, and convulsive activities. In bovine serum albumin. The percentage inhibition of addition, it is the major source of shikimic acid, a protein denaturation was calculated as follows. primary ingredient in the anti-flu drug (Tamiflu).[6] Percentage inhibition = 100 - ((O.D of test - O.D of Nonsteroidal anti-inflammatory drugs (NSAIDS) product control)/O.D of Control) × 100 are drugs reduce inflammation and relieve fever and pain by blocking enzymes and proteins made by The half maximal inhibitory concentration (IC50) the body. NSAIDs such as ibuprofen and naproxen value was defined as the concentration of the sample block a protein called prostaglandin. They also extract to IC50 of protein denaturation under the assay reduce swelling and inflammation caused by an condition. injury or a disease such as arthritis. NSAIDs are strong medicines. The actions they take in the RESULTS body to help one condition can cause problems Denaturation of proteins is a well-documented cause in other ways. NSAIDs may increase the risk of of inflammation. The anti-inflammatory activity of heart attack, stroke, skin reactions, and serious the extract was studied by its ability to inhibit protein stomach and intestinal bleeding. These risks are denaturation. It was effective in inhibiting heat induced greater if taken NSAIDs at higher doses or for albumin denaturation at different concentrations as longer periods than recommended. Some available shown in Table 1. Maximum inhibition, 77.87 ± 1.55 anti-inflammatory drugs are aspirin, diclofenac, was observed at 500 µg/ml. IC50 value was found ibuprofen, oxaprozin, etc.[7] to be 105.35 ± 1.99 µg/ml. Aspirin, a standard anti- The side effects of commercially available drugs: inflammatory drug showed the maximum inhibition, Nausea, vomiting, diarrhea, constipation, etc. 77.12 ± 1.42% at the concentration of 200 µg/ml. Advances of using herbal anti-inflammatory are Values are means of three independent analyses of the they have no side effects. The human body’s natural sample ± standard deviation. response to injury results in inflammation-induced pain, swelling, and erythema. To reduce pain, anti- DISCUSSION inflammatory agents such as NSAIDs act on the multiple inflammatory pathways, which, although Star anise is one of many spices that contain often very effective, can have undesirable side bioactive compounds as well as a number of phenolic effects such as gastric ulceration and, infrequently, and flavonoid compounds, having antioxidant, myocardial infarction, and stroke. For centuries, preservative, and antimicrobial properties. It also natural anti-inflammatory compounds have been has many other properties of anti-inflammatory, used to mediate the inflammatory process and often antioxidant, and antifungal properties. This plant with a fewer side effect. Ongoing experiments and I. verum does contain anti-inflammatory property. clinical trials are being continued to guide and provide From the table, we can see that the plant showed their scientifically based effectiveness to reduce maximum inhibition of 77.87 ± 1.55 for 500 mg/ml. inflammation and promote wellness. The IC50 value was found to be 300.99 ± 1.99. Aspirin, also known as acetylsalicylic acid, is a medication MATERIALS AND METHODS used to treat pain, fever, or inflammation. Specific inflammatory conditions in which aspirin is used I. verum extract obtained commercially was used include Kawasaki disease, pericarditis, and rheumatic for the study. The anti-inflammatory activity of was fever.[8] Aspirin showed 80.12 ± 1.42 in 250 mg/ml. studied using protein denaturation assay. The IC50 value being 160.78 ± 0.50. Inhibition of Protein Denaturation (Kumar et al., Aspirin is a NSAID used in various pathological 2011) conditions for its anti-inflammatory, antipyretic, and The reaction mixture (0.5 ml) consisted of 0.45 ml analgesic benefits. For many years, aspirin and NSAIDs bovine serum albumin (5% aqueous solution) and have been used to address acute maladies (e.g., pain, 0.05 ml of the methanol extract of turmeric (100– fever, and inflammation).[9] More recently, increasing 500 μg/ml). pH was adjusted to 6.3 using a small knowledge of NSAIDs’ preventive effects against amount of 1N HCl. The samples were incubated at development and progression of cardiovascular diseases 37°C for 20 min and then heated at 57°C for 3 min. and cancer has encouraged their use for prevention of After cooling the samples, 2.5 ml phosphate buffer chronic diseases, as well as for acute pain.[10,11] However, 2442 Drug Invention Today | Vol 10 • Issue 12 • 2018 Rachel Paul and R. V. Geetha Table 1: Protein denaturation inhibiting activity of I. vercum extract Sample Percentage Control (aspirin) Percentage concentration (µg) activity % concentration (µg) activity % 100 12.59±1.05 50 17.97±0.50 200 34.65±1.75 100 32.68±0.57 300 48.95±0.65 150 47.39±1.50 400 61.63±1.15 200 63.07±1.49 500 77.87±1.55 250 80.12±1.42