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Effect of Eugenol on stomach Eradication of Helicobacter pylori is an important objective in overcoming gastric diseases. Many regimens are currently available but none of them could achieve 100% success in eradication. Eugenol and cinnamaldehyde that are commonly used in various food preparations are known to possess antimicrobial activity against a wide spectrum of bacteria.The present study was performed to assess the in vitro effects of eugenol and cinnamaldehyde against indigenous and standard H. pylori strains, their minimum inhibitory concentrations (MICs) and time course lethal effects at various pH.A total of 31 strains (29 indigenous and one standard strain of H. pylori ATCC 26695, one strain of E. coli NCIM 2089) were screened. Agar dilution method was used for the determination of drug sensitivity patterns of isolates to the commonly used antibiotics and broth dilution method for the test compounds.Results These results indicate that the two bioactive compounds we tested may prevent H. pylori growth in vitro, without acquiring any resistance.1 Administration of natural or synthetic agents to inhibit, delay, block, or reverse the initiation and promotional events associated with carcinogenesis opens a new avenue for cancer prevention and treatment to reduce cancer morbidity and mortality. Eugenol, a potential chemopreventive agent, is a component of clove and several other spices such as basil, cinnamon, and bay leaves. A number of reports have shown that eugenol possesses antiseptic, analgesic, antibacterial, and anticancer properties. The present study was undertaken to evaluate the chemopreventive potential of eugenol alone and in combination with a chemotherapeutic agent such as gemcitabine. Eugenol showed dose-dependent selective cytotoxicity toward HeLa cells in comparison to normal cells, pointing to its safe cytotoxicity profile. A combination of eugenol and gemcitabine induced growth inhibition and apoptosis at lower concentrations, compared with the individual drugs. The analysis of the data using a combination index showed combination index values of <1 indicating strong synergistic interaction. The combination thus may enhance the efficacy of gemcitabine at lower doses and minimize the toxicity on normal cells. In addition, the expression analysis of genes involved in apoptosis and inflammation revealed significant downregulation of Bcl-2, COX-2, and IL-1β on treatment with eugenol. Thus, the Results suggest that eugenol exerts its anticancer activities via apoptosis induction and anti-inflammatory properties and also provide the first evidence demonstrating synergism between eugenol and gemcitabine, which may enhance the therapeutic index of prevention and/or treatment of cervical cancer.2 Clove (Syzygium aromaticum) is one of the mostcommonly used spices in Indian kitchens. It has

beenshown to be a potent chemo preventive agent, used by thetraditional Ayurvedic healers of India since ancient timesto treat respiratory and digestive ailments (Banerjee et al., 2006). Essential oils present in the dried flower buds ofclove are eugenol, caryophyllene, alphahumulene, alphaterpinyl acetate, eugenyl, methyl eugenol, actyl eugenol,naphthalene, chavicol, heptanone, sesquiterpenes, (Duke and Cellier, 1993) Spices, active ingredients of Indian cooking, may play important roles in prevention and treatment of variouscancers. The objective of the present study is to compare the in vitro anticancer activities of three different extracts of Clove (Syzygium aromaticum L), a commonly used spice and food flavouring agent, against different kinds of cancer cell lines of various anatomical derivations. Water, ethanol and oil extracts were screened for anti proliferative activity against HeLa (cervical cancer), MCF-7 (ER + ve) and MDA-MB-231 (ER – ve) breast cancer, DU-145 prostate cancer and TE-13 esophageal cancer cell lines, along with normal human peripheral blood lymphocytes. Inhibition of cell proliferation was assessed using MTT assay as a vital stain. In the examined fve cancer cell lines, the extracts showed different patterns of cell growth inhibition activity, with the oil extract having maximal cytotoxic activity. Morphological analysis and DAPI staining showed cytotoxicity to be a result cell disruption with subsequent membrane rupture. Maximum cell death and apoptotic cell demise occurred in TE-13 cells within 24 hours by clove oil at 300µl/ml with 80% cell death whereas DU-145 cells showed minimal cell death. At the same time, no signifcant cytotoxicity was found in human PBMC’s at same dose.Result eugenol present in clove oil extract is an effective cytotoxic agent for different type of cancer cells and it is endowed with apoptotic inducing capability. These results suggest that eugenol may constitute a potential antitumor compound against different kind of cancer cells depending up on their sensitivity towards it.3 Loss of function of the p53 gene is implicated in defective apoptotic responses of tumors to chemotherapy. Although the pro-apoptotic roles of eugenol and capsaicin have been amply reported, their dependence on p53 for apoptosis induction in gastric cancer cells is not well elucidated. The aim of the study was to elucidate the role of p53 in the induction of apoptosis by eugenol and capsaicin in a human gastric cancer cell line, AGS cells were incubated with or without various concentrations of capsaicin and eugenol for 12 hrs, in the presence and absence of p53 siRNA. Cell cycling, annexin V and expression of apoptosis related proteins Bax, Bcl-2 ratio, p21, cyt c-caspase-9 association, caspase-3 and caspase-8 were studied.In the presence of p53, capsaicin was a more potent pro-apoptotic agent than eugenol. However, silencing of p53 significantly abrogated apoptosis induced by capsaicin but not that by eugenol. Western blot analysis of pro-apoptotic markers revealed that as opposed to capsaicin, eugenol could induce caspase-8 and caspase-3 even in the absence of p53. Result: Unlike capsaicin, eugenol could induce apoptosis both in presence and absence of functional p53. Agents which can induce apoptosis irrespective of the cellular p53 status have immense scope for development as potential anticancer agent.4

Essential oils present in the dried flower buds ofclove are eugenol, caryophyllene, alpha-humulene, alphaterpinyl acetate, eugenyl, methyl eugenol, actyl eugenol, naphthalene, chavicol, heptanone, sesquiterpenes.Among different essential oils eugenol is the principle component, present in amount of 81.1%.Beside this transcryophyllene and isoeugenol are present in amount of 7% and 10.1% respectively (Zheng et al.,1992).The main compound of clove is eugenol. Spices, active ingredients of Indian cooking, may play important roles in prevention and treatment of various cancers. The objective of the present study is to compare the in vitro anticancer activities of three different extracts of Clove (Syzygium aromaticum L), a commonly used spice and food flavoring agent, against different kinds of cancer cell lines of various anatomical derivations. Water, ethanol and oil extracts were screened for anti proliferative activity against HeLa (cervical cancer), MCF-7 (ER + ve) and MDA-MB-231 (ER – ve) breast cancer, DU-145 prostate cancer and TE-13 esophageal cancer cell lines, along with normal human peripheral blood lymphocytes. Inhibition of cell proliferation was assessed using MTT assay as a vital stain. In the examined five cancer cell lines, the extracts showed different patterns of cell growth inhibition activity, with the oil extract having maximal cytotoxic activity. Morphological analysis and DAPI staining showed cytotoxicity to be a result of cell disruption with subsequent membrane rupture. Maximum cell death and apoptotic cell demise occurred in TE-13 cells within 24 hours by clove oil at 300µl/ml with 80% cell death whereas DU-145 cells showed minimal cell death. At the same time, no significant cytotoxicity was found in human PBMC’s at the same dose.Result: In conclusion this work demonstrates that the eugenol present in clove oil extract is an effective cytotoxic agent for different type of cancer cells and it is endowed with apoptotic inducing capability. These results suggest that eugenol may constitute a potential antitumor compound against different kind of cancer cells depending up on their sensitivity towards it.5

Possible mechanisms underlying the gastroprotective effect of eugenol against indomethacin‐ induced ulcer in rats were investigated. Pyloric ligation was performed for collection of gastric juice, and gastric ulceration was induced by a single intraperitoneal (i.p.) injection of indomethacin (30 mg/kg). Pretreatment with a single dose of eugenol (100 mg/kg, orally), 1 h

before indomethacin administration caused significant reductions in gastric mucosal lesions, gastric acid outputs and pepsin activity associated with a significant increase in mucin concentration. Additionally, eugenol significantly attenuated the elevations in gastric mucosal malondialdehyde and total nitrite, and the decrease in reduced glutathione observed with indomethacin. The protective effect afforded by eugenol was significantly inhibited by prior administration of glibenclamide, the ATP‐sensitive potassium (KATP) channel blocker, but not by prior use of ruthenium red, the transient receptor potential vanilloid 1 (TRPV1) antagonist. The results indicate that the anti‐ulcer effect of eugenol is mediated by opening of KATP channels, scavenging free radicals, decreasing acid‐pepsin secretion, increasing mucin production, and preventing the deleterious rise in nitric oxide level.6

Syzygium aromaticum, a medicinal plant commonly known as clove, is used to treat toothache, respiratory disorders, inflammation, and gastrointestinal disorders. From the flower buds of S. aromaticum, it is possible to obtain an essential oil comprised of a mixture of aliphatic and cyclic volatile terpenes and phenylpropanoids, being eugenol as the main component. The aims of this study were: (1) to extract the essential oil of the flower buds of S. aromaticum, (2) to identify and quantify the main component of the essential oil, and (3) to evaluate its antiulcer activity using different animal models. Assays were performed using the following protocols in rats: indomethacin-induced and ethanol/HCl-induced ulcer model. Both essential oils from S. aromaticum and eugenol displayed antiulcer activities in the rat models of indomethacin- and ethanol-induced ulcer. Studies focusing on the possible mechanisms of gastroprotection were also undertaken using the following experiments: evaluation of gastric secretion by the pylorus-ligated model, determination of mucus in gastric content, participation of nitric oxide (NO) and endogenous sulfhydryl in gastric protection. The results show that there was no significant effect on the volume of gastric juice and total acidity. However, the quantification of free gastric mucus showed that the clove oil and eugenol were capable of significantly enhancing mucus production. With regard to the NO and endogenous sulfhydryls, the results demonstrated that the gastroprotection induced by clove oil and eugenol are not related to the activities of the nitric oxide and endogenous sulfhydryls. No sign of toxicity was observed in the acute toxicity study. In conclusion, the results of this study show that essential oil of S. aromaticum, as well as its main component (eugenol), possesses antiulcer activity. The data suggest that the effectiveness of the essential oil and eugenol is based on its ability to stimulate the synthesis of mucus, an important gastroprotective factor. However, further pharmacological and toxicological investigations are required to enable its use for the treatment of gastric ulcer. 7

References: 1. Ali SM, Khan AA, Ahmed I, Musaddiq M, Ahmed KS, Polasa H, Rao LV, Habibullah CM, Sechi LA, Ahmed N. Antimicrobial activities of Eugenol and Cinnamaldehyde against the human gastric pathogen Helicobacter pylori. Annals of clinical microbiology and antimicrobials. 2005 Dec;4(1):20.

2. Hussain A, Brahmbhatt K, Priyani A, Ahmed M, Rizvi TA, Sharma C. Eugenol enhances the chemotherapeutic potential of gemcitabine and induces anticarcinogenic and anti-inflammatory activity in human cervical cancer cells. Cancer Biotherapy and Radiopharmaceuticals. 2011 Oct 1;26(5):519-27. 3. Dwivedi V, Shrivastava R, Hussain S, Ganguly C, Bharadwaj M. Comparative anticancer potential of clove (Syzygium aromaticum)—an Indian spice—against cancer cell lines of various anatomical origin. Asian Pac J Cancer Prev. 2011 Jan 4.

https://www.ncbi.nlm.nih.gov/pubmed/?term=anti+cancer+role+of+eugenol+on+stomach

5. Dwivedi V, Shrivastava R, Hussain S, Ganguly C, Bharadwaj M. Comparative anticancer potential of clove (Syzygium aromaticum)—an Indian spice—against cancer cell lines of various anatomical origin. Asian Pac J Cancer Prev. 2011 Jan 1;12(8):198993. 6. Morsy MA, Fouad AA. Mechanisms of gastroprotective effect of eugenol in indomethacin-induced ulcer in rats. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2008 Oct;22(10):1361-6. 7. Santin JR, Lemos M, Klein-Júnior LC, Machado ID, Costa P, de Oliveira AP, Tilia C, de Souza JP, de Sousa JP, Bastos JK, de Andrade SF. Gastroprotective activity of essential oil of the Syzygium aromaticum and its major component eugenol in different animal models. Naunyn-Schmiedeberg's archives of pharmacology. 2011 Feb 1;383(2):149-58.