PENGARUH PEMBERIAN VITAMIN D AKTIF PADA MENCIT JANTAN DENGAN UNILATERAL URETERAL OBSTRUCTION Kajian terhadap Skor Cedera Tubulus, Jumlah Makrofag M1 & M2, serta Jumlah Myofibroblast

RIZKA ADI N. P., dr. Nur Arfian, Ph.D.; dr. Santosa Budiharjo, M. Kes., P.A(K).

2017 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Pendahuluan: Chronic Kidney Disease (CKD) merupakan salah satu masalah kesehatan yang menjadi isu global dan memberikan prognosis yang buruk berupa terjadinya fibrosis ginjal. Unilateral Ureteral Obstruction (UUO) merupakan metode eksperimen paling representatif untuk mendapatkan model fibrosis ginjal. Fibrosis ginjal ditandai dengan adanya cedera progresif pada jaringan parenkim ginjal serta adanya penumpukan matriks ekstraseluler, berupa kolagen tipe I dan III sebagai penanda khasnya. Penelitian ini bertujuan untuk mengetahui pengaruh pemberian vitamin D aktif terhadap jumlah makrofag M1 dan M2, skor cedera tubulus ginjal, serta jumlah myofibroblast, pada mencit jantan dengan UUO. Metode: Dua puluh lima mencit jantan galur Swiss usia 3 bulan (30-40 gram)dibagi menjadi 5 kelompok, yaitu perlakuan, kelompok UUO3, UUO7, UUOD3 dan UUOD7. Kelompok UUO mendapatkan etanol 0,2% sedangkan kelompok UUO+Vit.D mendapatkan vitamin D dengan dosis 0,5 mcg/kgBB selama 3 hari dan 7 hari. Cedera tubulus diamati dengan menggunakan pewarnaan Periodic AcidSchiff, pengamatan makrofag M1 melalui immunohistokimia CD 68, makrofag M2 melalui immunohistokimia Arginase I, dan myofibroblast melalui immunohistokimia +-SMA. Skor cedera tubulus, jumlah makrofag dan myofibroblast dihitung dengan software Image J, kemudian data dianalisis menggunakan software SPSS22. Hasil: Perlakuan UUO menyebabkan terjadinya peningkatan cedera tubulus ginjal, jumlah makrofag M1, jumlah myofibroblast dan penurunan jumlah makrofag M2 seiring dengan bertambahnya durasi perlakuan (p<0,05). Setelah dilakukan pemberian vitamin D, terjadi penurunan skor cedera tubulus ginjal, jumlah makrofag M1 dan jumlah myofibroblast, serta peningkatan jumlah makrofag M2 dibandingkan dengan kelompok UUO (p<0,05). Kesimpulan: Pemberian vitamin D aktif menghambat pembentukan fibrosis ginjal melalui penurunan cedera tubulus ginjal, rasio makrofag M1/M2, serta penurunan jumlah myofibroblast.

Introduction. Chronic Kidney Disease (CKD) is the global health issues with bad prognosis of kidney fibrosis behavioural. Unilateral Ureteral Obstruction (UUO)is the most representative experimental methodology to obtain kidney fibrosis model. Kidney fibrosis observed with progressive injury of parenchymal kidney and extracellular matrix aggregation of type I and III collagen. This study was aimed to explore the effect of active vitamin D administration toward the expression of macrophage M1 and M2, tubular injury and myofibroblast in male mice by using UUO. Methods. Twenty-five Switzerland furrowed mouse were divided into 5 equals groups; control group (SO), UUO3, UUO7, UUOD3 and UUOD7. UUO groups received 0.2% ethanol and UUOD groups received 0.5 mcg/kg BM vitamin D for 3 and 7 days exposure. Twenty-five paraffin-embedded section of kidney tissue were analysed by Periodic Acid Schiff and immunohistochemical staining against antibody anti-CD68, Arginase I and +-SMA. Data were shown in the proportion of tubular injury, M1 and M2 macrophage, and myofibroblast accumulation with imageJ software for scoring. Data were analysed using one way anova using SPSS22 software with p<0.05 considered to be statistically significant. Results. UUO treatment increased the tubular injury, M1 macrophage and myofibroblast accumulation but decreased M2 macrophage (p<0.05) compared to SO group. This study also revealed the poorer prognosis in different exposure of treatment (p<0.05). But vitamin D (UUOD) administration to the treatment decreased the tubular injury, M1 macrophage and myofibroblast accumulation and increased M2 macrophage (p<0.05) compared to UUO groups. Conclusion. Active vitamin D administration decreased kidney fibrosis by reducing tubular injury and M1-M2 macrophage ratio and inducing myofibroblast in mice.

Kata Kunci : Unilateral Ureteral Obstruction, Makrofag M1, Makrofag M2, Rasio, Cedera tubulus, Myofibroblast, Unilateral Ureteral Obstruction, M1-M2 Macrophage ratio, Tubular Injury