Uji Diagnostik Biomarker Trombopoietin, TGF- Beta1, Angiotensin II Untuk Penentuan Derajat Fibrosis Pada Hepatitis Kronik Studi Pada Tikus Rattus norvegicus strain Sprague-Dawley

WAHYU SISMINDARI, Prof. dr. Siti Nurdjanah, SpPD(KGEH), MKes ; dr. Totok Utoro, DMedSc, SpPA(K); Prof. dr. Budi Mulyono, MM, SpPK(K)

2015 | Disertasi | S3 Ilmu Kedokteran

Sirosis hepatis merupakan penyakit hati kronik yang ditandai dengan pembentukan jaringan fibrosis dan nodul-nodul regeneratif. Fibrosis hati adalah akumulasi jaringan matriks ekstraseluler yang menyebabkan perubahan molekuler dan histologi hati. Baku emas diagnosis fibrosis hati adalah biopsi hati. Namun, biopsi hati mempunyai beberapa kendala dan kelemahan sehingga perlu dicari alternatif pemeriksaan salah satunya dengan memeriksa biomarker non invasif. Pemeriksaan non invasif yang selama ini banyak digunakan masih memberikan variabilitas hasil yang luas. Penelitian ini bertujuan mencari nilai diagnostic trombopoetin (TPO), transforming growth factor Beta1 (TGF-Beta1) dan angiotensin II dalam memprediksi derajat fibrosis. Penelitian ini menggunakan desain cross sectional. Perlakuan berupa pemberian CCl4 secara intra peritoneal kepada 48 tikus Rattus norvegicus strain Sprague-Dawley jantan, umur 2 bulan, berat badan 220 - 249 gram, yang terbagi dalam 6 kelompok. Kelompok P0 mendapatkan olive oil selama 12 minggu. Kelompok P1 mendapatkan injeksi CCl4 selama 1 minggu, P2 selama 4 minggu, P3 selama 6 minggu, P4 selama 8 minggu, dan P5 selama 12 minggu, dengan dosis 0.2 ml/100 gram BB (2x/minggu) selama 2 minggu dilanjutkan 0.1 ml/100 gram BB sampai akhir masa perlakuan. Selanjutnya, dilakukan pemeriksaan TPO, TGF-Beta1 dan angiotensin II darah, serta histopatologi hati. Hasil penelitian menunjukkan adanya hubungan bermakna antara kadar TPO dengan derajat fibrosis (r=-0.329, p=0.026). Tidak terdapat hubungan antara kadar TGF-Beta1 dengan derajat fibrosis hati (r=0.169, p=0.262). Didapatkan hubungan bermakna antara kadar kadar angiotensin II dengan derajat fibrosis hati (r=0.309, p=0.037). Nilai diagnostik TPO dalam penentuan derajat fibrosis hati adalah sensitivitas 0.811, spesifisitas 0.667, nilai duga positif (NDP)0.909, nilai duga negatif (NDN)0.461, rasio likelihood postif 2.45, dan rasio likelihood negative 0.28. Nilai diagnostik TGF-Beta1 didapatkan sensitivitas 0.865, spesifisitas 0.333, NDP 0.842, NDN 0.375, rasio likelihood positif 1.30 dan rasio likelihood negative 0.41. Sedangkan nilai diagnostik angiotensin II adalah sensitivitas 0.973, dengan spesifisitas 0.333, NDP 0.857, NDN 0.75, rasio likelihood positif 1.46 dan rasio likelihood negatif 0.08. Kombinasi parameter terbaik adalah antara TPO + TGF-Beta1 + angiotensin II dengan titik potong skor 3 yang menghasilkan sensitivitas 0.70 dengan spesifisitas 0.89, NDP 0.96, NDN 0.42, rasio likelihood positif 6.36 dan rasio likelihood negatif 0.34. Hasil penelitian ini mendapatkan bahwa trombopoetin, TGF-Beta1 dan angiotensin II, baik sebagai biomarker tunggal maupun kombinasi, lebih tepat digunakan sebagai pemeriksaan skrining untuk penentuan derajat fibrosis.

Hepatic cirrhosis is a chronic liver disease characterized by the formation of fibrosis and regenerative nodules. Liver fibrosis is the accumulation of extracellular matrix tissue which causes the molecular and histological changes in the liver. Gold standard diagnosis of liver fibrosis is liver biopsy. However, liver biopsy has several constraints and weaknesses, so we need to look for an alternative examination by using non-invasive biomarkers. Non-invasive examinations that have been widely used still have a wide variability of results. Objective of this research is to find the diagnostic values of thrombopoietin (TPO), transforming growth factor beta 1 (TGF-Beta1)and angiotensin II in predicting the degree of fibrosis. This study was conducted using a cross-sectional design. Treatment in the form of intraperitoneal administration of CCl4 to 48 Rattus norvegicus Sprague-Dawley strain, male, 2 month, weight 220 – 249 gram, were divided into 6 groups. P0 group was given olive oil for 12 weeks. P1 group got CCl4 injection for 1 week, P2 for 4 weeks, P3 for 6 weeks, P4 for 8 weeks, and P5 for 12 weeks, with dose 0.2 ml/100g BW (2x/week)for 2 weeks then 0.1 ml/100g BW until the end of the treatment. Blood was collected for examination of TPO, TGF-Beta1, angiotensin II, and the liver was taken for histopathology examination. The results showed a significant relationship between TPO levels and the degree of fibrosis (r=-0.329, p=0.026). There was no correlation between levels of TGF-Beta1 and the degree of liver fibrosis (r=0.169, p=0262), but there was a significant correlation between the levels of angiotensin II and the degree of liver fibrosis (r=0309, p=0.037). Thrombopoietin diagnostic values in determining the degree of liver fibrosis are sensitivity of 0.811, specificity of 0.667, positive predictive value (PPV)of 0.909, negative predictive value (NPV)of 0.461, positive likelihood ratio of 2.45, and the negative likelihood ratio of 0.28. The diagnostic values of TGF-Beta1 showed a sensitivity of 0.806, specificity of 0.333, PPV of 0.842, NPV of 0.375, a positive likelihood ratio of 1.30 and a negative likelihood ratio of 0.41. While the diagnostic values of angiotensin II are sensitivity of 0.973, specificity of 0.333, PPV of 0.857, NPV of 0.75, a positive likelihood ratio of 1.46 and a negative likelihood ratio of 0.08. The best combination is TPO + TGF-Beta1 + angiotensin I parameters with cut-off point score 3, showed sensitivity of 0.70, specificity of 0.89, PPV of 0.96, NPV of 0.42, positive likelihood ratio of 6.36 and the negative likelihood ratio of 0.34. This study found that thrombopoietin, TGF-Beta1 and angiotensin II, either as a single or combination of biomarkers, are more appropriately used as a screening test for the determination of the degree of fibrosis

Kata Kunci : nilai diagnostik, trombopoetin, TGF-Beta1, angiotensin II, derajat fibrosis, tikus Rattus norvegicus strain Sprague-Dawley, diagnostic values, thrombopoietin, TGF-Beta1, angiotensin II, degree of fibrosis, Rattus norvegicus strain Sprague-Dawley