Tesis

EFEK PEMBERIAN SEL PUNCA MESENKIM ASAL TALI PUSAT MANUSIA PADA EKSPRESI DAN DISTRIBUSI OV6 DAN CK19 JARINGAN HATI TIKUS SETELAH INDUKSI CCL4 DAN 2-AAF = EFFECT OF HUMAN UMBILICAL CORD-DERIVED MESENCHYMAL STEM CELLS ON OV-6 AND CK-19 EXPRESSION AND DISTRIBUTION IN RAT LIVER AFTER CCL4 AND 2-AAF INDUCTION.

Sel punca mesenkim asal tali pusat manusia terbukti bermanfaat pada regenerasi hati saat terjadi cedera kronis. Namun, masih perlu penelitian lebih mendalam mengenai peran pemberian sel punca mesenkim asal tali pusat manusia (SPM-TP) pada regenerasi hati akibat cedera kronis melalui sel oval (sel progenitor hati). Penelitian ini bertujuan untuk melihat apakah pemberian SPM-TP pada model tikus dengan cedera kronis akibat Carbon tetrachloride (CCl4) dan 2-Acetylaminofluorene (2-AAF) selama 12 minggu dapat meningkatkan ekspresi dan distribusi sel oval melalui pemeriksaan imunohistokimia OV6 dan CK19. Jaringan sampel didapatkan dari bahan biologi tersimpan penelitian sebelumnya, yaitu lima kelompok (n=6) tikus (Rattus novergicus) strain Wistar jantan usia 8 minggu yang diberikan CCl4 dan 2-AAF selama 12 minggu. Kelompok satu dan dua adalah kelompok kontrol dan induksi 2-AAF/CCl4 yang dieuthanasia pada akhir minggu ke-12, sedangkan kelompok tiga, empat, dan lima adalah kelompok yang dieuthanasia pada akhir minggu ke-14 dan terdiri dari kelompok kontrol, induksi 2-AAF/CCl4 serta induksi 2-AAF/CCl4 ditambah pemberian SPM-TP. Ekspresi OV6 dan CK19 dinilai secara kuantitatif melalui analisis skor optical density dari program IHC Profiler-ImageJ. dari gambar digital pada tiap zona hati. Hasilnya, ekspresi OV6 dan CK19 gabungan seluruh zona hati menunjukkan peningkatan yang bermakna antara kelompok SPM-TP dengan kelompok kontrol 14 minggu (p=0,004 dan 0,016). Sedangkan pengamatan ekspresi OV6 berdasarkan zona hati menunjukkan peningkatan ekspresi OV6 yang bermakna (p=0,006) antara kelompok SPM-TP terhadap kelompok kontrol dan induksi 14 minggu di zona I dan II. Pengamatan ekspresi CK19 dari gabungan seluruh zona hati menunjukkan peningkatan secara bermakna antara kelompok SPM-TP dengan kelompok kontrol 14 minggu (p=0,0016). Sedangkan ekspresi CK19 pada zona II menunjukkan peningkatan ekspresi CK19 yang bermakna antara kelompok SPM-TP dengan kelompok induksi dan kontrol 14 minggu (p=0,028 dan 0,006). Dengan demikian, terdapat perbedaan ekspresi OV6 dan CK19 pada kelompok SPM-TP dibandingkan dengan kelompok induksi 14 minggu. Distribusi ekspresi OV6 paling banyak di zona I dan CK19 paling banyak di zona II hati tikus dengan pemberian SPM-TP. Perbedaan ekspresi dan distribusi ekspresi OV6 dan CK19 pada kelompok tikus yang diberikan SPM-TP menunjukkan perbedaan aktivasi dan proliferasi sel oval.
Keywords: Regenerasi hati, 2-AAF/CCl4, Sel oval, Sel punca mesenkim asal tali pusat manusia, OV6, CK19


Human umbilical cord mesenchym stem cells (hUC-MSCs) have been shown to be beneficial in liver regeneration during chronic injury. However, more in-depth research is needed on the role of HUC-MSCS in liver regeneration due to chronic injury through oval cells (known as liver progenitor cells). The study aimed to see if administering HUC-MSCS in mouse models with chronic injury from Carbon tetrachloride (CCl4) and 2-Acetylaminofluorene (2-AAF) for 12 weeks could improve oval cell expression and distribution through immunohistochemistry (IHC) examinations of OV6 and CK19. The sample tissue was obtained from biological material stored by previous research. Five groups (n =6) of rat (Rattus novergicus) strains of 8-week-old male Wistar were given CCl4 and 2-AAF for 12 weeks. Groups one and two were the 2-AAF/CCl4 control and induction group that was euthanized at the end of week 12, while groups three, four, and five were the group that were euthanized at the end of week 14 and consisted of a control group, 2-AAF/CCl4 induction and 2-AAF/CCl4 induction plus HUCMSCS administration. The expressions of OV6 and CK19 were assessed quantitatively through the optical density score analysis of the IHC Profiler-ImageJ program. from digital images in each liver zone. As a result, OV6 and CK19 expressions of the entire liver zone data showed a significant increase between the HUC-MSCS group and the 14-week control group (p=0.004 and 0.016). Observations of OV6 expressions based on the liver zone showed a noteably increase in OV6 expression (p=0.006) between the HUC-MSCS group against the control group and a 14-week induction in zones I and II. CK19 expression from the entire liver zone showed a significant increase between the hUC-MSCs group and the 14-week control group (p=0.0016). Furthermore, the CK19 expression in zone II showed a significant increase in CK19 expression between the HUC-MSCS group and the 14-week induction and control group with the p=0.028 and 0.006 respectively. Thus, there were differences in OV6 and CK19 expressions in the hUC-MSCs group compared to the 14-week induction group. The distribution of OV6 expression was mostly in zone I and CK19 was mostly in zone II of rat liver with the administration of HUC-MSCS. Differences in expression and distribution of OV6 and CK19 expressions in the group of rats hUC-MSCs-induced group showed differences in oval cell activation and proliferation.
Keywords: Liver regeneration, 2-AAF/CCl4, Oval cells, Human umbilical mesenchym stem cells, OV6, CK19