Pengaruh Pemberian Probiotik Escherichia Coli Strain Nissle 1917 dan Prebiotik Sereal Beras Hitam (Oryza sativa L. ‘Sembada Hitam’) Terhadap Tingkat Kerusakan Histologi Kolon serta Distribusi Makrofag Pada Mencit Model Colitis Associated Colorectal Cancer (CA-CRC)

solikhah, Dr. Yekti Asih Purwestri, S.Si., M.Si.; Dewi Kartikawati Paramita, S.Si, M.Si, Ph.D

2024 | Tesis | S2 Biologi

Colitis-associated colorectal cancer (CA-CRC) merupakan hasil akumulasi mutasi pada sel epitel usus dan berkembang menjadi peradangan kronis yang ditandai dengan displasia hingga adanya kejadian yang mendahului kanker kolorektal. Penyakit radang usus (inflammatory bowel disease/IBD) IBD) merupakan faktor risiko dalam perkembangan kanker kolorektal terkait kolitis (CA-CRC). Pemberian prebiotik sereal beras hitam (BRC) dan/ atau probiotik Escherichia coli Nissle 1917 (EcN berpotensi memiliki efek imunomodulator dan aktivitas antioksidan. Penelitian ini bertujuan untuk mengetahui tingkat kerusakan histologis, tingkat distribusi makrofag, tingkat indeks aktivitas penyakit (IAP) pada jaringan kolon setelah pemberian perlakuan prebiotik sereal beras hitam (BRC) dan/atau probiotik Escherichia coli Nissle 1917 (EcN). Metode penelitian ini menggunakan tiga puluh lima ekor mencit BALB/c dibagi menjadi 7 kelompok perlakuan termasuk kelompok kontrol. Induksi azoxymethane (AOM) 10 mg/kg berat badan diberikan secara injeksi intraperitoneal, dilanjutkan dengan pemberian DSS 2?lam air minum sebanyak 3 siklus. Kelompok mencit terdiri dari kelompok kontrol normal (KK), kelompok CA-CRC atau kelompok induksi tanpa perlakuan (KP1), kelompok CA-CRC dengan prebiotik awal-akhir (KP2), kelompok prebiotik CA-CRC dengan probiotik awal-akhir (KP3), kelompok CA-CRC dengan probiotik akhir (KP4), kelompok sinbiotik awal-akhir (KP5), kelompok sinbiotik akhir (KP6). Gambaran tingkat kerusakan histopatologi diamati pada irisan jaringan kolon FFPE dengan pewarnaan HE, distribusi makrofag M1 dengan imunohistokimia antibodi anti-iNOS, dan makrofag M2 dengan anti-CD163. Selain itu juga dilakukan scoring indeks aktivitas penyakit (IAP). Hasil: Kerusakan histologi pada semua kelompok berbeda signifikan terhadap kelompok CA-CRC (KP1) (p<0>

Background Colitis-associated colorectal cancer (CA-CRC) is consecuences of accumulated mutations in intestinal epithelial cells and develops into chronic inflammation, which characterised by dysplasia until the presence of events that precede colorectal cancer, Inflammatory bowel disease (IBD) is a risk factor for colitis-associated colorectal cancer (CA-CRC). The administration of probiotic Escherichia coli Nissle 1917 (EcN) and prebiotic black rice cereals (BRC) with have potential immunomodulatory effects and antioxidant activity. Objectives The aim of this study is to investigat the level of histological damage, macrophage distribution and disease activity index (DAI) in colonic tissue after the treatment of black rice cereal prebiotics (BRC) and or probiotic Escherichia coli Nissle 1917 (EcN). Methods Thirty-five BALB/c mice were divided into 7 groups including a control group. The induction of azoxymethane (AOM) 10 mg/kg body weight was given by intraperitoneal injection, followed by the administration of 2% DSS in drinking water for 3 cycles. The groups consisted of normal control group (KK), CA-CRC group or induction group without treatment (KP1), CA-CRC group with prebiotics from early-late (KP2), CA-CRC group with probiotics early-late (KP3), CA-CRC group with probiotics late (KP4), early-late synbiotic group (KP5), late synbiotic group (KP6).  Histopathological damage level was observed in FFPE colon tissue slices with HE staining, M1 macrophage distribution was examined with anti-iNOS antibody immunohistochemistry, and M2 macrophages with anti-CD163. Disease activity index (IAP) scoring was also performed. Results Histological damage in all groups was significantly different from the CA-CRC group (KP1) (p<0.05). The mildest colonic histological damage was observed in the early-late synbiotic group (KP5), followed by the late synbiotic group (KP6) and in both groups the histological picture almost resembled the normal control group (KK). The highest distribution of M1 macrophages was found in the CA-CRC group (KP1) with an IRS score of 10.5. Followed by other groups, namely KP2 and KP3 (IRS score of 6.5), KP4 with a score of 3, and KP5 and KP6 (IRS score of 1.5). M2 macrophages were not found in the KK, KP4 and KP6 groups (score 0). The distribution of M2 macrophages was mostly found in group KP1 (CA-CRC) and group KP2 (early-late prebiotics) (score 2). KP3 (early-late probiotic) and KP5 (early-late synbiotic) groups showed lower M2 macrophage distribution (score 0.5). The early-late synbiotic group (KP5) had the lowest final IAP picture to approach 0, followed by groups KP2, KP3, KP4 and KP6 compared to group KP1 (CA-CRC) (p<0.05). Conclusions The combination of BRC and EcN synbiotics gave the mildest histopathological damage effect, the distribution of M1 and M2 macrophages in the group given BRC prebiotics and/or EcN probiotics was lighter in CA-CRC mice. The disease activity index also appeared to be the mildest in the group treated with BRC and EcN synbiotics.
Keywords: CA-CRC, Black Rice Cereal (BRC), EcN 1917,  macrophages.

Kata Kunci : Colitis-associated Colorectal Cancer (CA-CRC), Black Rice Cereal (BRC), macrophages.