adyu j sci Adıyaman University Journal of Science 2147-1630 2146-586X Adıyaman University 10.37094/adyujsci.697784 Structural Biology Yapısal Biyoloji LCMSMS Analysis of 8-OHdG and Measuring Metallothionein Level for Evaluation of Protective Role of Geraniol in Lead Acetate Administered Rats https://orcid.org/0000-0002-8957-7476 Alkan Uçkun Aysel ADIYAMAN ÜNİVERSİTESİ https://orcid.org/0000-0002-9730-9471 Yoloğlu Ertan ADIYAMAN ÜNİVERSİTESİ https://orcid.org/0000-0002-9018-8515 Uçkun Miraç ADIYAMAN ÜNİVERSİTESİ https://orcid.org/0000-0002-0173-3084 Özkaya Ahmet ADIYAMAN ÜNİVERSİTESİ 06 25 2020 10 1 100 111 03 03 2020 05 29 2020 Copyright © 2011, Adıyaman University Journal of Science 2011 Adıyaman University Journal of Science

In this study, DNA damage and metallothionein levels were used as biomarkers to evaluate the protective potential of geraniol, a monoterpen against lead stress, in rats. Hepatic 8-hydroxy-2-deoxyguanosine (8-OHdG) level used as a marker of oxidative DNA damage was measured by LCMSMS. Experimental groups were formed in four ways: control, lead acetate, geraniol and lead acetate + geraniol. Seven animals were used in each group. Geraniol and lead acetate were administered to rats for 30 days. In geraniol administered rats, 8-OHdG and metallothionein levels decreased significantly compared to lead acetate administered rats. The highest DNA damage and metallothionein levels were observed in lead acetate administered rats. According to the results of this study, it can be suggested that geraniol protects cells against lead-caused damage by reducing ROS production. In addition, studies on the measurement of 8-OHdG by LCMSMS in the literature are limited. Therefore, it is thought that the presented study will contribute to the evaluation of the applicability of this method in the literature.

 8-hydroxy-2-deoxyguanosine Metallothionein Geraniol Lead LCMSMS Adıyaman Üniversitesi FEFYL/2015–0005 [[1] Sinicropi, M.S., Amantea, D., Caruso, A., Saturnino, C., Chemical and biological properties of toxic metals and use of chelating agents for the pharmacological treatment of metal poisoning, Archives of Toxicology, 84, 501-520, 2010. [2] Carocci, A., Rovito, N., Sinicropi, M.S., Genchi, G., Mercury toxicity and neurodegenerative effects, Reviews of Environmental Contamination and Toxicology, 229, 1-18, 2014. [3] Silbergeld, E.K., Waalkes, M., Rice, J.M., Lead as a carcinogen, experimental evidence and mechanisms of action, American Journal of Industrial Medicine, 38, 316-323, 2000. [4] Adonaylo, V.N., Oteiza, P.I., Pb2+ promotes lipid peroxidation and alteration in membrane physical properties, Toxicology, 132, 19-32, 1999. [5] Garcia, M.T.A., Gonzalez, E.L.M., Toxic effects of perinatal lead exposure on the brain of rats, involvement of oxidative stress and the beneficial role of antioxidants, Food Chemistry and Toxicology, 46, 2089-2095, 2008. [6] Jayachandran, M., Chandrasekaran, B., Namasivayam, N., Geraniol attenuates fibrosis and exerts anti-inflammatory effects on diet induced atherogenesis by NF-κB signaling pathway, European Journal of Pharmacology, 762, 102-111, 2015. [7] Barnard, D.R., Xue, R., Laboratory evaluation of mosquito repellents against Aedes albopictus, Culex nigripalpus, and Ochlerotatus triseriatus (Diptera, Culicidae), Journal of Medical Entomology, 41, 726-730, 2004. [8] Burke, Y.D., Stark, M.J., Roach, S.L., Sen, S.E., Crowell, P.L., Inhibition of pancreatic cancer growth by the dietary isoprenoids farnesol and geraniol. Lipids, 32, 151-156, 1997. [9] Bard, M., Albrecht, M.R., Gupta, N., Guynn, C.J., Stillwell, W., Geraniol interferes with membrane functions in strains of Candida and Saccharomyces, Lipids, 23, 534-538, 1988. [10] Hamer, D.H., Metallothionein, Annual Review of Biochemistry, 55, 913-951, 1986. [11] Takahashi, S., Molecular functions of metallothionein and its role in hematological malignancies, Journal of Hematology and Oncology, 5, 41, 2012. [12] Cherian, M.G., Jayasurya, A., Bay, B.H., Metallothioneins in human tumors and potential roles in carcinogenesis, Mutation Research, 533, 201-209, 2003. [13] Datta, J., Majumder, S., Kutay, H., Motiwala, T., Frankel, W., Costa, R., Cha, H.C., MacDougald, O.A., Jacob, S.T., Ghoshal, K., Metallothionein expression is suppressed in primary human hepatocellular carcinomas and is mediated through inactivation of CCAAT/enhancer binding protein alpha by phosphatidylinositol 3-kinase signaling cascade, Cancer Research, 67, 2736-2746, 2007. [14] Yan, D.W., Fan, J.W., Yu, Z.H., Li, M.X., Wen, Y.G., Li, D.W., Zhou, C.Z., Wang, X.L., Wang, Q., Tang, H.M., Peng, Z.H., Downregulation of metallothionein 1F, a putative oncosuppressor, by loss of heterozygosity in colon cancer tissue, Biochimica et Biophysica Acta, 1822, 918-926, 2012. [15] Wei, H., Desouki, M.M., Lin, S., Xiao, D., Franklin, R.B., Feng, P., Differential expression of metallothioneins (MTs) 1, 2, and 3 in response to zinc treatment in human prostate normal and malignant cells and tissues, Molecular Cancer, 7, 7, 2008. [16] Zhang, B., Satoh, M., Nishimura, N., Suzuki, J.S., Sone, H., Aoki, Y., Tohyama, C., Metallothionein deficiency promotes mouse skin carcinogenesis induced by 7,12-dimethylbenz[a]anthracene, Cancer Research, 58, 4044-4046, 1998. [17] O'Donovan, P., Perrett, C.M., Zahang, X., Montaner, B., Xu, Y.Z., Harwood, C.A., McGregor, J.M., Walker, S.L., Hanaoka, F., Karran, P., Azathioprine and UVA light generate mutagenic oxidative DNA damage, Science, 309, 1871-1874, 2005. [18] Alan, M.R., Wang, Y., Shah, J., Gordon, S., Fager, M., Butter, PP., Jun, KH., Guardiola-Salmeron, C., Carabe-Fernandez, A., Fan, Y., Proton beam radiation induces DNA damage and cell apoptosis in glioma stem cells through reactive oxygen species, Scientific Reports, 5, 13961-13972, 2015. [19] Krystona, T.B., Georgieva, A.B., Pissisb, P., Georgakilasa, A.G., Role of oxidative stress and DNA damage in human carcinogenesis, Mutation Research, 711, 193-201, 2011. [20] Dizdaroglu, M., Oxidatively induced DNA damage and its repair in cancer. Mutation Research, 763, 212-245, 2015. [21] Cooke, M.S., Evans, M.D., Dizdaroglu, M., Lunec, J., Oxidative DNA damage, mechanisms, mutation, and disease, Faseb Journal, 17, 1195-1214, 2003. [22] Gajewski, E., Rao, G., Nackerdien, Z., Dizdaroglu, M., Modification of DNA bases in mammalian chromatin by radiationgenerated free radicals, Biochemistry, 29, 7876-7882, 1990. [23] Shimoda, R., Nagashima, M., Sakamoto, M., Yamaguchi, N., Hirohashi, S., Yokota, J., Kasai, H., Increased formation of oxidative DNA damage, 8-hydroxydeoxyguanosine, in human livers with chronic hepatitis, Cancer Research, 54, 3171-3172, 1994. [24] Guo, C., Li, X., Wang, R., Yu, J., Ye, M., Mao, L., Zhang, S., Zheng, S., Association between Oxidative DNA damage and risk of colorectal cancer, Sensitive Determination of Urinary 8-Hydroxy-2-deoxyguanosine by UPLC-MS/MS Analysis, Scientific Reports, 6, 32581, 2016. [25] Ozkaya, A., Sahin, Z., Kuzu, M., Saglam, Y.S., Ozkaraca, M., Uckun, M., Yologlu, E., Comakli, V., Demirdag, R., Yologlu, S., Role of geraniol against lead acetate-mediated hepatic damage and their interaction with liver carboxylesterase activity in rats, Archives of Physiology and Biochemistry, 124, 80-87, 2017. [26] Viarengo, A., Ponzano, E., Dondero, F., Fabbri, R., A simple spectrophotometric method for metallothionein evaluation in marine organisms, an application to Mediterranean and Antarctic molluscs, Marine Environmental Research, 44, 69-84, 1997. [27] Hsu, P.C., Guo, Y.L., Antioxidant nutrients and lead toxicity, Toxicology, 180, 33-44, 2002. [28] Kasai, H., Increased formation of oxidative DNA damage, 8-hydroxydeoxyguanosine, in human livers with chronic hepatitis, Cancer Research, 54, 3171-3172, 1994. [29] Dribben, W.H., Creeley, C.E., Farber, N., Low-level lead exposure triggers neuronal apoptosis in the developing mouse brain, Neurotoxicology and Teratology, 33, 473-480, 2011. [30] Courtois, E., Marques, M., Barrientos, A., Casado, S., López-Farré, A., Lead-induced downregulation of soluble guanylate cyclase in isolated rat aortic segments mediated by reactive oxygen species and cyclooxygenase-2, Journal of the American Society of Nephrology, 14, 1464-1470, 2003. [31] Bolin, C.M., Basha, R., Cox, D., Zawia, N.H., Maloney, B., Lahiri, D.K., Cardozo-Pelaez, F., Exposure to lead and the developmental origin of oxidative DNA damage in the aging brain, Faseb Journal, 20, 788-790, 2006. [32] Xu, J., Lian, L.J., Wu, C., Wang, X.F., Fu, W.Y., Xu, L.H., Lead induces oxidative stress, DNA damage and alteration of p53, Bax and Bcl-2 expressions in mice, Food Chemistry and Toxicology, 46, 1488-1494, 2008. [33] Liu, C.M., Ma, J.Q., Sun, Y.Z., Puerarin protects the rat liver against oxidative stress mediated DNA damage and apoptosis induced by lead, Experimental Toxicology and Pathology, 64, 575-582, 2012. [34] Wang, C., Liang, J., Zhang, C., Bi, Y., Shi, X., Shi, Q., Effect of ascorbic acid and thiamine supplementation at different concentrations on lead toxicity in liver, Annals of Occupational Hygiene, 51, 563-569, 2007. [35] Tiwari, M., Kakkar, P., Plant derived antioxidants-geraniol and camphene protect rat alveolar macrophages against t-BHP induced oxidative stress, Toxicology in Vitro, 23, 295-301, 2009. [36] Ong, T.P., Heidor, R., Conti, A.D., Dagli, M.L.Z., Moreno, F.S., Farnesol and geraniol chemopreventive activities during the initial phases of hepatocarcinogenesis involve similar actions on cell proliferation and DNA damage, but distinct actions on apoptosis, plasma cholesterol and HMGCoA reductase, Carcinogenesis, 27, 1194-1203, 2006. [37] Chen, C.Y., Jhou, Y.T., Lee, H.L., Lin, Y.W., Simultaneous, rapid, and sensitive quantification of 8-hydroxy-2'-deoxyguanosine and cotinine in human urine by on-line solid-phase extraction LC-MS/MS, correlation with tobacco exposure biomarkers NNAL, Analytical and Bioanalytical Chemistry, 408, 6295-6306, 2016. [38] Li, C.S., Wu, K.Y., Gou-Ping., Chang-Chien, Chou, C.C., Analysis of oxidative DNA damage 8-Hydroxy-2'-deoxyguanosine as a biomarker of exposures to persistent pollutants for marine mammals, Environmental Science and Technology, 39, 2455-2460, 2005. [39] Dai, S., Yin, Z., Yuan, G., Yu, H., Jia, R., Xu, J., Song, X., Li, L., Shu, Y., Liang, X., He, C., Lv, C., Zhang, W., Quantification of metallothionein on the liver and kidney of rats by subchronic lead and cadmium in combination, Environmental Toxicology and Pharmacology, 36, 1207-1216, 2013. [40] Ikebuchi, H., Teshima, R., Suzuki, K., Simultaneous induction of Pb-metallothionein-like protein and Zn-thionein in the liver of rats given lead acetate, Biochemical Journal, 233, 541-546, 1986. [41] Gillis, B.S., Arbieva, Z., Gavin, I.M., Analysis of lead toxicity in human cells, BMC Genomics, 13, 344, 2012. [42] Nakao, K., Kibayashi, K., Taki, T., Koyama, H., Changes in the Brain after Intracerebral Implantation of a Lead Pellet in the Rat, Journal of Neuroscience, 27, 1925-1934, 2010. [43] Chidinma, N.C., Adewale, A., Chiaka, A., Differential expression of metallothionein-1 and cytochrome p450 2a5 (cyp2a5) in mice in response to lead acetate exposure and industrial effluents in Ibadan, Nigeria, Toxicology and Industrial Health, 32, 1975-1981, 2016. [44] Wang, J., Su, B., Zhu, H., Chen, C., Zhao, G., Protective effect of geraniol inhibits inflammatory response, oxidative stress and apoptosis in traumatic injury of the spinal cord through modulation of NF-jB and p38 MAPK, Experimental and Therapeutic Medicine, 12, 3607-3613, 2016. [45] Prasad, S.N., Muralidhara, M., Protective effects of geraniol (a monoterpene) in a diabetic neuropathy rat model, attenuation of behavioral impairments and biochemical perturbations, Journal of Neuroscience Research, 92, 1205-1216, 2014.