Malaysian Applied Biology Journal

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29-1&2-09

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Malays. Appl. Biol. (December 2000) 29(1&2): 69-74

THE ROLE OF EXTRACELLULAR CA2+ ON THE INHIBITORY EFFECTS OF ACETYLCHOLINESTERASE ACTIVITY IN HARUAN, CHANNA STRIATUS BLOCK, BRAIN TISSUE BY HEAVY METALS

MAT-JAIS, A.M. and MOHAMED, M.Z.

Department of Biomedical, Faculty of Medicine and Health Sciences, 43400 University Putra Malaysia, Serdang, Selangor, Malaysia.

ABSTRACT

In this study, we are investigating the in vitro inhibition of acetylcholinesterase activity in Channa striatus brain tissue :v mercury, cadmium, lead, nickel and zinc and the role of extracellular calcium. Fresh brain tissue was cut out from 35.50 ± 3.20 crn in length, 150.00 + 2.50 gm in weight cultured C.striatus, and homogenised in a phosphate buffer (0.1M, pH 7.0) (1:4 w/v) using a biohomogeniser (Biospees Product Inc.), M133/1281-0 at 4°C. The homogenised sample was then centrifuged for 20 mins. at 10,000 g using Beckman GS-6R centrifuge the resultant supernatant used for the assay. Acetylcholinesterase activity in C. striatus brain tissue was measured following the Ellman, et al., 1961 method using Shimadzu UP-2401PC spectrophotometer. Protein was determined using Lowry, et al., 1951 method to calculate the acetylcholinesterase specific activity. The sample was exposed to a series of in vitro sub-lethal concentration of each metal 2.50; 5.00; 7.50; 10.00 and 20.00 ppm were exposed to the assay. All the heavy metals studied had inhibited acetylcholinesterase activity in C. striatus brain tissue in a dose dependent manner. The inhibition were found to be significantly different at p < 0.05. Mercury was the most toxic, with 2.50 ppm enough to suppress 30 % of the acetylcholinesterase activity. It is therefore, based on the concentration of 20.00 ppm, we found that Hg>Cd>Pb>Zn>Ni in toxicity, at which mercury inhibited 180.76 ± 25.94 %; cadmium 148.08 ± 5.36 %; lead 88.19 ± 1.19 %; zinc 62.43 ± 2.28 % and nickel 64.19 ± 2.38 %. Subsequently, 20.00 ppm extracellular calcium induced 5.02 ±4.12 % stimulation of the acetylcholinesterase activity. It is also interesting to note that the lower concentrations of extracellular calcium 2.50 and 5.00 ppm had increased the acetylcholinesterase activity to 17.04 + 2.69 and 23.36 + 2.69 % respectively.


ABSTRAK

Di dalam kajian ini, kami melihat perencatan aktiviti asetilkolinesterase dalam tisu otak Channa striatus secara in vitrc oleh logam berat raksa, kadmium, plumbum, nikel dan zinkum, dan peranan kalsium ekstrasel. Tisu otak segar diperolehj dari C. striatus ternakan berukuran 35.50 + 3.20 cm, 150.00 ± 2.50 gm, dan dihomogenkan dalam penimbal fosfal 0.1M, pH 7.0, 1:4 (b/i) menggunakan biohomogeniser (Biospees Product Inc.), M133/1281-0 pada 4°C. Sampel yang slab dihomogen, kemudiannya diempar seterusnya selama 20 min pada 10,000 g menggunakan pengempar Beckman iS-6R. Aktiviti asetilkolinesterase dalam tisu otak C. striatus telah diukur mengikut kaedah Edman, et at., 1961 r.nsgunakan spectrofotometer Shimadzu UP-2401PC, dan protein ditenfukan berdasarkan kepada kaedah Lowry, et '., 1951 untuk menentukan aktiviti spesifik asetilkolinesterase. Sampel tisu otak C. straitus tela didedahkan kepada reberapa siri kepekatan separa-maut in vitro setiap logam iaitu 2.50; 5.00; 7.50; 1.00 and 20.00 ppm. Kesemua logam boat didapati merencat aktiviti asetilkolinesterase dalam tisu otak C. striatus. Perencatan logam berat ini dipengaruhi dos logam dan mempunyai perbezaan ketara pada p < 0.05. Raksa adalah paling toksik dengan dos 2.50 ppm yang ;_kup untuk merencat sebanyak 30% aktiviti asetilkolinesterase. Oleh itu, berdasarkan kepada kepekatan 20.00 ppm, kami mendapati bahawa ketoksikan logam berat terhadap C. striatus adalah Hg>Cd>Pb>Zn>Ni, di mana raksa telah erencat sebanyak 180.76 + 25.94 %; diikuti oleh kadmium 148.08 + 5.36 %; plumbum 88.19 + 1.19 %; zinkum 1 -3 ± 2.28 % dan nikel 64.19 + 2.38 %. Berikutnya, 20.00 ppm kalsium ekstrasell telah memulihkan 5.02 ± 4.12 -~ rangsangan terhadap aktiviti acetylcholinesterase. Seterusnya, adalah menarik untuk diperhatikan bahawa kepekatan talsium ekstra-sellular yang lebih rendah 2.50 dan 5.00 ppm telah meningkatkan aktiviti asetilkolinesterase kepada 17.04 - 2.69 dan 23.36 + 2.69 % masing - masing.

Key words: heavy metal, acetylcholinesterase activity, inhibition, and extracellular calcium

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