PERBANDINGAN DETEKSI PROLIFERASI SEL DENGAN IMUNOHISTOKIMIA PCNA DAN BrdU PADA PANKREAS TIKUS

David Pakaya, Wiwit Setyowati, Rina Susilowati

DOI: https://doi.org/10.23917/biomedika.v12i2.10641

Abstract

ABSTRAK

Pankreas merupakan jaringan labil karena terdiri dari sel-sel asinar yang selalu berproliferasi sepanjang hidupnya untuk menggantikan sel yang rusak. Proliferasi tersebut dapat diidentifikasi dengan pemeriksaan histologis, yaitu mikroteknik dengan pewarnaan imunohistokmia (IHC). Penelitian ini bertujuan untuk membandingkan deteksi proliferasi sel dengan IHC anti-PCNA dan anti-BrdU pada pankreas. Tikus diinjeksi BrdU secara intraperitoneal sebanyak 4 kali dengan selang waktu 3 jam dengan dosis BrdU 0,125 ml, 0,375 ml, 0,250 ml, dan 0,250 ml dan dibiarkan selama 1 malam. Jaringan pankreas dipotong sepanjang ± 5 mm dan direndam dalam fiksatif paraformaldehid 4%, kemudian dibuat blok paraffinnya. Blok parafin diiris dengan ketebalan 6µm untuk pengecatan hematoksilin eosin (HE) dan 4 µm untuk pengecatan IHC. Pengecatan IHC dengan antibodi anti PCNA (antibodi monoklonal PCNA 1;1000 dalam PBS). Pengecatan IHC dengan antibodi anti BrdU menggunakan antibodi primer (antibodi anti BrdU 1:1000). Pengecatan dianalisis secara kualitatif. IHC menggunakan antibodi anti-PCNA dan anti-BrdU menunjukkan adanya penyebaran warna coklat gelap yang menandakan proliferasi sel aktif, dimana sel yang terekspresi pada BrdU lebih sedikit dibandingkan PCNA.  Penelitian ini menyimpulkan bahwa proliferasi sel pankreas dapat terekspresi melalui perwarnaan IHC dengan antibodi anti PCNA dan anti BrdU.

Kata kunci: Imunohistokimia, PCNA, Brdu, Proliferasi

 

ABSTRACT

                   The pancreas is a labile tissue because it consists of acinar cells which proliferate throughout their lives to replace damaged cells. The proliferation can be identified by histological examination, that is microtechnics with immunohistochmia (IHC) staining. This study aims to compare the detection of cell proliferation with anti-PCNA and anti-BrdU IHC in the pancreas. The rats were injected BrdU intraperitoneally 4 times with an interval of 3 hours with a dose of BrdU 0.125 ml, 0.375 ml, 0.250 ml, and 0.250 ml and left for 1 night. Pancreatic tissue were necropsy along ± 5 mm and soaked in 4% paraformaldehyde fixative, then made paraffin block. The paraffin block was sliced to a thickness of 6 µm for hematoxylin eosin (HE) and 4 μm for IHC painting. IHC used anti PCNA antibodies (PCNA monoclonal antibody 1; 1000 in PBS) and anti BrdU antibodies (anti BrdU primary antibodies 1: 1000). Staining was analyzed qualitatively. IHC with anti-PCNA and anti-BrdU antibodies showed the spread of dark brown which indicates active cell proliferation, whereas cells expressed in BrdU are less than PCNA. This study concluded that pancreatic cell proliferation can be expressed through the coloring of IHC with anti-PCNA and anti-BrdU antibodies.

Keyword: Immunohistochemistry, PCNA, Brdu, Proliferation

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