ABSTRAKRuang lingkup dan cara penelitian: Telah dilakukan penelitian induksi kanker hati tikus dengan aflatoksin B1. Penelitian ini ingin melihat apakah asam sialat meningkat lebih dini dari AFP pada induksi kanker hati tersebut. 50 ekor tikus perlakuan diinduksi dengan intubasi lambung 42 x 20 ug aflatoksin B1 Sebagai kontrol, 50 ekor tikus diintubasi dengan sejumlah sama akuades. Pengamatan dilakukan dengan mengambil plasma dan jaringan hati setiap bulan selama 10 bulan pemeliharaan. Pada contoh uji dilakukan pengukuran kadar asam sialat, deteksi AFP plasma dan pengamatan histopatologis jaringan hati. Pengukuran kadar asam sialat dilakukan dengan cara spektrofotometri, sedangkan deteksi AFP dengan cara ELISA. Pengukuran AFP dengan teknik ELISA memerlukan antigen AFP dan antibodi anti AFP tikus, yang ternyata tidak tersedia dipasaran. Antibodi anti AFP dibuat dengan imunisasi kelinci menggunakan protein amnion. Setelah imunisasi ke 3 titer antibodi sebesar 1280, setelah imunisasi ke 5 sebesar 2560. Purifikasi antibodi dilakukan dengan kolom imunoafinitas AminoLink. Purifikasi AFP dari amnion dilakukan dengan kolom afinitas Cibacron Blue. Uji statistik yang dipakai untuk analisis hasil adalah analisis Marian dua arah, dengan batas kemaknaan p<0,05.Hasil dan kesimpulan: Hasil penelitian menunjukkan bahwa kadar asam sialat plasma kelompok tikus perlakuan berbeda bermakna dengan kelompok kontrol sejak bulan pertama (p<0,05). Deteksi AFP menunjukkan peningkatan AFP pada kelompok tikus perlakuan dan tidak pada kelompok tikus kontrol sejak bulan pertama (p<0,05). Pengamatan histopatologis menunjukkan adanya perubahan yang dapat menuju kearah keganasan. Dengan demikian hasil pengamatan petanda tumor pada tikus yang diinduksi kanker hati menunjukkan bahwa baik kadar AFP maupun asam sialat meningkat pada waktu dini, satu bulan setelah induksi dengan aflatoksin B1 .
ABSTRACTDetermination of The Earliness of Rat Plasma α-Fetoprotein (AFP) and Sialic Acid Levels as Tumor Markers in the Process of Liver Carcinogenesis Induction By Aflatoxin B1 (AFB1) Scope and method of study: An experimental study of aflatoxin B1 induced liver carcinogenesis has been done. The aim oh this study was to prove the increase of plasma sialic acid level is earlier than the AFP level in liver carcinogenesis. Fifty rats that were treated with daily intubations of 20 gg for a period of 42 days. Fifty control rats were intubated with equal volumes of aquadest. Observation of the plasma and liver tissues were done every month, including measurement of plasma AFP, sialic acid level, and histopatology of the liver tissue. Sialic acid level was done by spectrophotometric technique, and ELISA technique was used for the detection of plasma AFP. The ELISA technique for rat AFP needs rat AFP antigen and antibody anti rat AFP which were not available from chemical suppliers. The antibody anti rat AFP was developed by immunization of rabbits with rat amniotic protein. After the 3rd immunization, rabbit antibody anti rat amniotic titter was 1280, and after the 5th immunization 2560. Antibody purification was done by immunoaffinity chromatography column (AminoLink). Purification of the rat AFP from amniotic fluid was done by affinity chromatography column (Cibacron Blue). The result were analyzed by Analysis of varians, with p<0,05.Results and conclusions: The results showed that the plasma sialic acid level of rats treated with aflatoxin B1 were significantly different from that of the control rats, p<0,05. Besides, plasma AFP was detected in the treated rats, but not in the control rats, from the first month. The histopathologic observation of the treated rat livers showed that there were changes that could lead to neoplastic livers. Both plasma and sialic acid levels were seen early, starting from the first month after induction of carcinogen by aflatoxin B1. |
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