Effects of Melissa officinalis L. on Oxidative Status and Biochemical...

[Full title: Effects of Melissa officinalis L. on Oxidative Status and Biochemical Parameters in Occupationally Exposed Workers to Aluminum: A Before after Clinical Trial]

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Life International Journal of Pharmacology ISSN 1811-7775 Life science alert ansinet Asian Network for Scientific Information

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International Jounal of Pharmacology 8 (5): 455-458, 2012 ISSN 1811-7775 DOI: 10.3923/ijp.2012.155.158 2012 Asian Network for Scientific Information Effects of Melissa officinalis L. on Oxidative Status and Biochemical Parameters in Occupationally Exposed Workers to Aluminum: A Before after Clinical Trial 'Davood Fazli, Ali Akbar Malekirad, Ali Asghar Pilevarian, 'Homayon Salehi, Akbar Zerralpisho, Kobra Rahzani and Mohammad Abdollahi 'Department of Biology, Payame Noor University. Tehran, Iran "Islamic Azad University, Jiroft Branch, Jiroft, Iran *Faculty of Neursing and Midwifery, Arak University of Medical Science, Arak, Iran *Faculty of Pharmacy and Pharmaceutical Sciences Research, Tehran University of Medical Science, Tehran, Iran Abstract: This study was designed to evaluate potential of Melissa officinalis L. (Lemon balm) infusion on the improvement of oxidative stress markers in workers occupationally exposed to aluminum (Al). In this beforeafter clinical trial, 30 Al workers were asked to drink Lemon balm infusion which was prepared like a tea bag twice per day (1.5 g/100 mL) for 30 days. Blood samples before and after entering the study were measured for lipid peroxidation (LPO), Total Antioxidant Capacity (TAC) Total Thiol Molecules (TTM), liver enzyme and some blood parameters. Use of Lemon balm infusion resulted in a significant increase in plasma levels of TTM and TAC and a significant decrease in triglyceride, cholesterol and aspartate transaminase (AST). LPO was not different before and after treatment. The conclusion is that infusions of Lemon balm improves oxidative stress condition in Al workers when used as a dietary supplement. Key words: Lemon balm, oxidative stress, aluminum, toxicity, workers INTRODUCTION Aluminium (Al) is a frequently found metal in the environment while most of people are not aware of the sources of Al. In fact, Al can be found in almost everywhere in food, drinking water, cosmetics, loothpaste and as adjuvant in different parenteral preparation and pharmaceutical agents. It has been shown that users of Al-containing antacids and buffered aspirin may have increased body Al (Krewski et al., 2007). Workers in the industries related to Al, are usually in a chronic exposure to Al more than that expected coming from normal daily diet. Our previous study showed that Al production workers who are occupationally exposed to Al have an oxidative stress situation that is evident in their blood (Ranjbar et al., 2008). As reviewed recently by Mohammadirad and Abdollahi (2011), most of toxicities of Al in human being are mediated through disturbing the balance between body oxidant and antioxidant. The most. common condition related to Al exposure is Alzheimer's Disease (AD). Al is known to induce or worsen AD by its oxidant effects (Garcia et al., 2010). With the same mechanism of action, Al is known as a risk factor for Parkinson's disease (Sanchez-Iglesias et al., 2009). Since oxidant/antioxidant imbalance is involved in the pathogenesis of many diseases (Abdollahi et al., 2004) thus it would not be surprising to find strong links between Al exposure and many deliberating diseases other than AD and Parkinson. Melissa officinalis L. (Lemon balm) belongs to the family Lamiaceae that grows in the Central and Souther Europe, Asia and Northen Iran. In Iran this plant is called locally as Badranjbooye, Varangboo and Faranjmoshk. Lemon balm contains a rich source of natural antioxidants and effective many oxidant-related disorders (Hasani-Ranjbar et al., 2009). A recent study indicated that Lemon balm is beneficial in protection against oxidative stress and DNA damage in subjects exposed to long-term low-dose ionizing radiation (eraatpishe et al.. 2011). hw Regarding above information, we aimed to evaluate possible benefit of Lemon balm on oxidative stress status of workers of an Al production factory who are chronically exposed to Al. MATERIALS AND METHODS Materials: The main chemicals used in this study were dithiobisnitrobenzoic acid (DTNB), Tris base 1,1.3.30tetraethoxypropane (MDA), 2-thiobarbituric acid (TBA), Corresponding Author: Mohammad Abdollahi, Faculty of Pharmacy and Pharmaceutical Sciences Research Center, Tehran University of Medical Science Tehran 1417614411, Tran 455

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Int. J. Pharmacol., 8(5): 455-458, 2012 trichloroacetic acid (TCA), n-butanol and 2,4,6-tripyridyls-triazine (TPTZ) that were obtained Merck Chemical Co. (Tehran). Plant material: The aerial parts of Melissa officinalis L. were collected in August 2009 from Botanical Garden of Shaheed Beheshti University and identified as Melissa officinalis L. by Dr. M.A. Vakili from Department of Biology, l'aculty of Science, Islamic Azad University, Jiroft Branch. The leaves of Melissa officinalis L. was dried in shade at room temperature for 12 days. were Study population: The study was conducted on 30 male workers, with age range of 22-65, who worked in the Al production factory located in an industrial part of the Iran in Arak province. The factory has started its activity since 1972 and the product of this factory is 175000 tons per year of all kinds of Al bar. Now about. 1700 workers are working in the factory. Al is produced by electrolysis of alumina (A10) in electrolytic cells (pol). Al is reduced by carbon from the Na AlF. The subjects occupationally exposed to Al by inhalation. All participants of the study before entering the study were provided with specific written information about the ais of the study and then were asked to give written consents in accordance with chical rules of Pharmaceutical Sciences Research Center (PSRC) of Tehran University of Medical Science (TUMS) where the study protocol was approved. Information on occupational history, socioeconomic status (salary, education) and lifestyle information (smoking, alcohol consumption, drug uses. consumption of vitamin or antioxidant supplements and dietary habits) were obtained by a questionaires and a direct interview with each worker by a trained interviewer. All subjects were submitted to complete clinical examination to detect any signs or symptoms of chronic diseases such as arterial hypertension heart failure, cancer, thyroid disturbance, asthma, diabetes and anemia. Tchividuals with chronic disease, alcohol consumption. antioxidant consumption and/or under drug treatment, or exposure to other toxic materials, radiation therapy, or substance abuse were excluded from the study. The included subjects were administered Lemon balm infusion (1.5 g/100 mL) twice per day for 30 days at 7.5 a.m. and 2 p.m. every day. Doses were obtained from our previous studies (Malckirad et al., 2012; Zeraatpisho et al., 2011). A supervisor carefully checked to make sure that the volunteers were taking infusion properly. Blood samples were collected from all subjects before using infusion and 12 h after the last dose of 30-day treatment with infusion. Plasma preparation: Blood samples were collected from all subjects before using Lemon balm infusion and 12 h aller the last dose of 30-day treatment with infusion. Five milliliter of heparinized blood were obtained and centrifuged at 3000 g for 30 min at 4°C to separate plasma. The plasma samples were stored at -80°C until analyzed. Infusion preparation and protocol: Lemon balm infusion was prepared according to a standard protocol. To 3 g of plant leaves, 200 mL of distilled water was added. The initial temperature of added water was 98°C. Infusion was left to stay at room temperature without additional heating. Infusion time was 30 min (Zeraatpishe et al., 2011). Measurement of plasma TAC: Plasma TAC was determined by measuring the ability of plasma to reduce Fe³ to Fe. The complex between Fe and TPTZ gives a blue color with absorbance al 593 nm that is measured by spectrophotometer. Measurement of plasma TTM: A volume of plasma (0.20 mL) was mixed with 0.6 mL of Tris-EDTA bulter (Tris base 0.25 M, EDTA 20M pH 8.2) and 40 µL of 10 mM of DTNB in methanol. The final volume of the reaction mixture was made up to 4 mL by adding 3.16 mL. of methanol. The test tube was capped and the color was developed for 15-20 min, followed by centrifugation at 3000 g for 10 min at ambient temperature. The absorbance of the supernatant was measured at 412 nm. Measurement of plasma IPO: LPO of plasma was determined by the reaction of TBA with MDA and other lipid peroxides. Briefly, plasma samples were mixed with TCA (20%) and the precipitate was dispersed in H₂SO, (0.05 M). TBA (0.2% in sodium sulfate) was added and heated for 30 min in a boiling water bath LPO adducts were extracted by n-butanol and absorbance was measured at 532 nm. Statistical analysis: All data were analyzed with stats direct 2.7.8. A paired t-test was used for statistical comparisons of biochemical parameters. Pearson correlation coefficient was used to study the association between variables. P values lower than 0.05 were considered statistically significant. RESULTS Table 1 shows demographic information of subjects recruited in the study. The average levels of subjects ages and years of employment were 35.8-5.6 and 15±5.7, respectively. 456

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Table 1: Demographic data of study subjects Int. J. Pharmacol., 8(5): 455-458, 2012 Employment Age Subjects Exposed workers (Mean±SD) Sex 35.8±5.6 (n=30) Male (n=30) (100%) Current. Strokers N=5 16.67% years (Mean SD) 15±5.7 Table 2: Plasma oxidative stress and hematological markers before and after treatment with Lemon balt p-value 0.015 0.0001 0.89 Parameters Before Aller TTM (mM) 0.150=0.086 0.198±0.068 TAC (nol L¯) 0.27-0.77 3.13+0.53 LPO (nmal mL-¹) 18.95 19.76 18.87116 ALT (UL) 16.3-6.8 16.8318 AST (UL) 31.5-10.41 21.7±8.1 Triglyceride (g L-¹) 180.4-122.1 179.77+47.37 146.6+81.1 144.4+30.46 0.61 0.0001 0.036 0.0001 Cholesterol (g L-¹) Values are as Mean±SD Table 2 shows the average levels of TAC, LPO, TTM. AST, ALT, triglyceride and cholesterol. The mean levels of TAC and TTM were significantly raised while the reduction of LPO was not statistically significant. The mean levels of AST, triglyceride and cholesterol significantly decreased after using infusion. No linear correlation was found between age or years of employment and plasma biochemical parameters. DISCUSSION Use of Lemon balm infusion in Al workers caused a significant increase in blood TAC, TTM and a significant. reduction in hepatic and lipid markers. The first think that comes to mind is that accumulation of Al in the workers induces free radicals and oxidative stress. Poor and improper protection tools seem to be the main reason for increased oxidative stress. In our examination of the mine and interview with workers. it was clear that they did not use proper masks and were not properly trained to use working cloth and gloves or to take shower regularly. The only safety tools that they used was paper masks. Though there were suitable bathrooms, the workers did not use them so often. However, in case of working cloth, gloves and shoes. more workers were inclined to use them. Our team in another study have proved that workers of this factory have higher level of blood Al and oxidative stress (Ranjbar et al., 2008) that explains existence of exposure lo Al In support of the present findings, an increase in the activities of AST and ALT and a decline in the activity of acid phosphatase (ACP) were previously reported (Yousef et al., 2007). The effect of Melissa officinalis L. extract on hyperlipidemic rats was also studied. Administration of Melissa officinalis T.. extract reduced total cholesterol, total lipid, ALT, AST and ALP levels in scrum and LPO levels in liver tissue, moreover increased glutathione levels in the tissue (Bolkent et al., 2005). Many in vitro and ex vivo studies have shown antioxidant activity of Melissa officinalis extracts but in vivo studies especially in human are rare. In vivo studdies just showed that Melissa officinalis .. extracl. could decrease LPO in rodents (Birdane et al., 2007) and in liver tissue of hyperlipidemic rats (Bolkent et al., 2005) and in radiology staff (Zeraatpishe et al., 2011). Melissa officinalis L. extract has been useful as rich source of antioxidants (Dastmalchi et al., 2008). The main identified in tea phenolic compounds which were infusion from Lemon balm were rosmarinic acid, luteolin 7-0-glucoside, quercetin 3-rutinoside, gallic acid, quercetin 3-0 galactoside and ferulic acid. Hence, it seems that Lemon balm, due to its antioxidant components and scavenging properties could increase the activity of antioxidant defense and decrease oxidative stress and triglyceride, cholesterol and AST in Al workers. CONCLUSION The oral administration of Lemon balm infusion may be useful for the protection of the Al worker from toxic effects of Al that is mediated through oxidative stress. These findings encourage pursuing further studies such as determination of the effect of other antioxidants in Al induced oxidative stress and searching for natural antioxidants for Al workers in long-term Al exposures. Anyway, if workers use suitable protective tools and take daily shower, the absorption and entrance of Al into body would be reduced. ACKNOWLEDGMENT Authors wish to thank workers who participated in This study. REFERENCES Abdollahi, M., A. Ranjbar S. Shachia, S. Nikfar and A. Rezaic, 2004. Pesticides and oxidative stress: A review. Med. Sci. Monit., 10: RA141-RA147. Birdane, Y.O., M.E. Buyukokuroglu, F.M. Birdane, M. Cemek and II. Yavuz, 2007. Anti-inflammatory and antinociceptive effects of Melissa officinalis L. in rodents. Rev. Med. Vet., 158: 75-81. Bolkent, S., R. Yanardag. O. Karabulut-Bulan and B. Yesilyaprak, 2005. Protective role of Melissa officinalis L. extract. on liver of hyperlipidemic rals: A morphological and biochemical study. J. Ethnopharmacol. 99: 391-398. 457

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Int. J. Pharmacol., 8(5): 455-458, 2012 Dastmalchi, K., H.J.D. Domana, P.P. Oinonena, Y. Darwis, I. Laakso and H. Raimo, 2008. Chemical composition and In vitro antioxidative activity of a lemon balm (Melissa officinalis L.) extract. LWT-Food Sci. Techmol. 41 391-400. Garcia, T., JL. Esparza, M.R. Nogues, M. Romeu, J.L.. Domingo and M. Gomez, 2010. Chemical composition and in vitro antioxidative activity of a lemon balm (Melissa officinalis L.) extract. Hippocampus, 20: 218-225. IIasani-Ranjbar, S., B. Larijani and M. Abdollahi, 2009. A systematic review of the potential herbal sources of future drugs effective in oxidant-related diseases. Inflamm. Allergy Drug Targets, 8: 2-10. Krewski, D., R.A. Yokel, F. Nichoer, D. Borchell and J. Cohen et al., 2007. IIuman health risk assessment for aluminium, aluminium oxide and aluminium hydroxide. J. Toxicol. Environ. Health B Crit. Rew., 10: 1-269. Malckitud A.A., M. Mojtabacc, M. Faghih, G. Vaczi and M. Abdollahi. 2012. Effects of the mixture of Melissa officinalis L. Cinnamomum zeylanicum and Urtica dinica on hepatic enzymes activity in patients with nonalcoholic fatty liver disease. Int. J. Pharmacol., (In Press). Mohammadirad, A. and M. Abdollahi, 2011. A systematic review on oxidant/antioxidant imbalance in aluminum Toxicity. Int. J. Pharmacol., 7: 12-21. Ranjbar, A, K. Khani-Jazani, A. Sedighi, F. JalaliMashayekhi, M. Ghazi-Khansari and M. Abdollahi. 2008. Alteration of body total antioxidant capacity and thiol molecules in human chronic exposure to aluminum. Toxicol. Environ. Chem., 90: 707-713. Sanchez-Iglesias, S., E. Mendez-Alvarez, J. IglesiasGonzalez, A. Munoz-Patio, I. Sanchez-Sellero, J.L. Labandeira-Garcia and R. Soto-Otero, 2009. Brain oxidative stress and selective behaviour of aluminium in specific areas of rat brain potential effects in a 6OIIDA-induced model of Parkinson's disease. J. Neurochem., 109: 879-888. Yousef, M.I., KI. Kamel M.I. El-Guendi and F.M. El-Demerdash, 2007. An in vitro study on reproductive Toxicity of aluminium chloride on rabbit. sperm: The protective role of some antioxidants. Toxicology, 239: 213-223. Zeraatpishe, A., S. Oryan, M.H. Bagheri. A A. Pilevarian A.A. Malekirad, M. Baeeri and M. Abdollahi, 2011. Effects of Melissa officinalis L. on oxidative status and DNA damage in subjects exposed to long-term low-dose ionizing radiation. Toxicol. Ind. Health, 27: 205-212. 458